Patent Publication Number: US-2022220446-A1

Title: Compositions and methods for organoid generation and disease modeling

Description:
CROSS REFERENCE TO RELATED APPLICATIONS 
     This application is a divisional of patent application U.S. Ser. No. 16/301,671, filed Nov. 14, 2018, which is a U.S. national stage application under 35 U.S.C. § 371 of International PCT Application No. PCT/US2017/034278, filed May 24, 2017, which claims priority to and benefit of U.S. Provisional Patent Application No. 62/341,461, filed May 25, 2016, the contents of all of which are hereby incorporated by reference herein in their entireties. 
    
    
     STATEMENT OF RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSORED RESEARCH 
     This invention was made with government support under Grant Nos. DK057978 and DK0909962 awarded by the National Institutes of Health. The government has certain rights in the invention. 
    
    
     BACKGROUND OF THE INVENTION 
     Although animal disease models can yield insight into the pathogenesis of diseases, drugs screened and selected using animal models often fail to be adopted in human patients. Because evolutionary biology, molecular biology, and genetic studies show animals and humans can profoundly differ, recapitulating human disease using human cells and generation of functional human organs is urgently needed. 
     SUMMARY OF THE INVENTION 
     As described below, the present invention features compositions and methods for generating an organoid, including a pancreatic islet organoid or a pancreatic organoid. 
     In one aspect, the invention provides a method of generating a pancreatic islet organoid, the method involving culturing an induced pluripotent stem cell (iPSC)-derived beta-like cell in a 3-dimensional matrix containing gellan gum, thereby generating a pancreatic islet organoid. 
     In another aspect, the invention provides a cell culture including an iPSC-derived beta-like cell in a three-dimensional matrix containing gellan gum. 
     In another aspect, the invention provides a cell culture including a human iPSC-derived beta-like cell, a human adipose-derived stem cell (hADSC), and a human umbilical vein endothelial cell (HUVEC) in a three-dimensional matrix containing gellan gum. 
     In various embodiments of any aspect delineated herein, the cell culture includes an adipose-derived stem cell and/or an endothelial cell. 
     In another aspect, the invention provides a pancreatic islet organoid containing an iPSC-derived beta-like cell, where the organoid is vascularized and exhibits glucose-stimulated insulin secretion (GSIS). 
     In another aspect, the invention provides a pancreatic islet organoid containing an iPSC-derived beta-like cell, an iPSC-derived alpha cell, an iPSC-derived delta cell, an iPSC-derived duct cell, an adipose-derived stem cell (hADSC), and an endothelial cell where the organoid is vascularized and exhibits glucose-stimulated insulin secretion (GSIS), KCl-stimulated insulin secretion, GLP-1 stimulated insulin secretion, somatostatin secretion, and glucagon secretion. 
     In a related aspect, the invention provides a non-human organism transplanted with the organoid of any aspect delineated herein. 
     In another aspect, the invention provides a method of identifying an agent that modulates pancreatic activity and/or treats a pancreatic disease involving contacting a candidate agent with a pancreatic islet organoid or a pancreatic organoid; and measuring an activity of the organoid contacted with the candidate agent, where the candidate agent is identified as an agent that modulates pancreatic activity and/or treats a pancreatic disease if the activity of the organoid is altered relative to a reference. 
     In another aspect, the invention provides a method of identifying an agent that modulates pancreatic activity and/or treats a pancreatic disease involving administering a candidate agent to a non-human subject transplanted with a pancreatic islet organoid or a pancreatic organoid; and measuring a pancreatic activity of the non-human subject, where the candidate agent is identified as an agent that modulates pancreatic activity and/or treats a pancreatic disease if the pancreatic activity of the non-human subject is altered relative to a reference. 
     In another aspect, the invention provides a method of treating a pancreatic disease in a subject involving transplanting a pancreatic islet organoid into the subject, where the pancreatic islet organoid contains an iPSC-derived beta-like cell, is vascularized, and exhibits glucose-stimulated insulin secretion (GSIS). 
     In another aspect, the invention provides a method of treating type 1 diabetes in a subject, involving transplanting a pancreatic islet organoid into the subject, where the pancreatic islet organoid contains an iPSC-derived beta-like cell, is vascularized, and exhibits glucose-stimulated insulin secretion (GSIS). 
     In another aspect, the invention provides a pancreatic islet organoid generated by culturing an induced pluripotent stem cell (iPSC)-derived beta-like cell in a 3-dimensional matrix containing gellan gum. 
     In another aspect, the invention provides a pancreatic organoid generated by culturing an induced pluripotent stem cell (iPSC)-derived beta-like cell and an iPSC-derived exocrine component cell in a 3-dimensional matrix containing gellan gum. 
     In another aspect, the invention provides a liver organoid generated by culturing an induced pluripotent stem cell (iPSC)-derived hepatocyte in a 3-dimensional matrix containing gellan gum. 
     In another aspect, the invention provides a heart organoid generated by culturing an induced pluripotent stem cell (iPSC)-derived cardiomyocyte in a 3-dimensional matrix containing gellan gum. 
     In another aspect, the invention provides an intestinal organoid generated by culturing an induced pluripotent stem cell (iPSC)-derived intestinal cell in a 3-dimensional matrix containing gellan gum. 
     In various embodiments of any aspect delineated herein, the method involves culturing the iPSC-derived beta-like cell with an adipose-derived stem cell and/or an endothelial cell. In various embodiments of any aspect delineated herein, the method involves culturing the iPSC-derived beta-like cell with an iPSC-derived alpha-like cell, an iPSC-derived delta-like cell, and/or an iPSC-derived duct-like cell. 
     In various embodiments of any aspect delineated herein, the pancreatic islet organoid contains an iPSC-derived alpha-like cell, an iPSC-derived delta-like cell, and/or an iPSC-derived duct-like cell. In various embodiments of any aspect delineated herein, the pancreatic islet organoid includes an adipose-derived stem cell and/or an endothelial cell. In various embodiments of any aspect delineated herein, the pancreatic islet organoid exhibits KCl-stimulated insulin secretion, GLP-1 stimulated insulin secretion, somatostatin secretion, and/or glucagon secretion. In various embodiments of any aspect delineated herein, the pancreatic islet organoid expresses one or more of the beta cell transcription factors Pdx1, MafA, Pax4, Pax6, NeuroD1, Nkx6-1, Gata6, and Foxa2. In certain embodiments, the pancreatic islet organoid contains an iPSC-derived beta-like cell, an iPSC-derived alpha cell, an iPSC-derived delta cell, an iPSC-derived duct cell, an adipose-derived stem cell (hADSC), and an endothelial cell, where the organoid is vascularized and exhibits glucose-stimulated insulin secretion (GSIS), KCl-stimulated insulin secretion, GLP-1 stimulated insulin secretion, somatostatin secretion, and glucagon secretion. In various embodiments of any aspect delineated herein, the pancreatic islet organoid is surrounded by an iPSC-derived exocrine component. In various embodiments, the iPSC-derived exocrine component expresses one or more of the markers PDX1, Nkx6-1, and Ptf1. 
     In various embodiments of any aspect delineated herein, the method involves inducing or mimicking a pancreatic disease in the organoid or non-human subject. In various embodiments of any aspect delineated herein, the disease is induced by contacting the organoid with or administering to the non-human subject one or more of the following agents: a free fatty acid (FFA), glucose, and cytokine. In various embodiments of any aspect delineated herein, the disease is mimicked by culturing the organoid with pancreatic cancer cells, stellate cells, and immune cells to create human pancreatic cancer microenvironment. In various embodiments of any aspect delineated herein, the pancreatic activity is one or more of insulin secretion, beta cell apoptosis, expression or activity of a NDUFA4, ESRRG, G6PC2, MDH1, LDHA, KCNK3, or MAFA polypeptide or polynucleotide, amylase secretion, apoptosis of an exocrine component, collagen synthesis, and stellate cell activation. In various embodiments, the non-human subject is also transplanted with a liver organoid. 
     In various embodiments of any aspect delineated herein, the candidate agent increases insulin secretion. In various embodiments of any aspect delineated herein, the candidate agent identified as an agent that modulates pancreatic activity is tested for the ability to treat a pancreatic disease. In various embodiments, the pancreatic disease is type 2 diabetes or pancreatic cancer. 
     In various embodiments of any aspect delineated herein, the organoid is an organoid according to any aspect delineated herein. 
     In various embodiments, the non-human organism is a mammal (e.g., a mouse). 
     In various embodiments of any aspect delineated herein, the pancreatic islet organoid contains an iPSC-derived alpha-like cell, an iPSC-derived delta-like cell, and/or an iPSC-derived duct-like cell. 
     In various embodiments of any aspect delineated herein, the pancreatic islet organoid contains an adipose-derived stem cell and/or an endothelial cell. 
     In various embodiments of any aspect delineated herein, a pancreatic disease is induced or mimicked in the subject. In particular embodiments, the pancreatic disease is type 1 diabetes or type 2 diabetes. In certain embodiments, the subject is a mammal (e.g., human). In various embodiments of any aspect delineated herein, the subject is administered an immunosuppressive agent. 
     In various embodiments of any aspect delineated herein, the liver organoid expresses one or more of the markers AFP, ALB, and Cyp3a7. In various embodiments of any aspect delineated herein, the liver organoid exhibits insulin signaling, insulin resistance by palmitic acids, and lipid accumulation. 
     In various embodiments of any aspect delineated herein, the heart organoid expresses one or more of the markers hMlc2a, hNkx2-5, alphaMHC and KCNQ1. In various embodiments of any aspect delineated herein, the heart organoid exhibits cardiac beating. 
     In various embodiments of any aspect delineated herein, the intestinal organoid expresses one or more of the markers CDX2, Muc2, and Lgr5. In various embodiments of any aspect delineated herein, the intestinal organoid exhibits budding in response to R-spondin. 
     In various embodiments of any aspect delineated herein, the iPSC-derived beta-like cell, iPSC-derived alpha-like cell, iPSC-derived delta-like cell, and/or iPSC-derived duct-like cell is human. In various embodiments of any aspect delineated herein, the iPSC-derived beta-like cell, iPSC-derived exocrine component cell, iPSC-derived hepatocyte, iPSC-derived cardiomyocyte, or iPSC-derived intestinal cell is human. In various embodiments, the adipose-derived stem cell is a human adipose-derived stem cell (hADSC). In various embodiments of any aspect delineated herein, the endothelial cell is a human umbilical vein endothelial cell (HUVEC). 
     In various embodiments of any aspect delineated herein, the pancreatic islet organoid, pancreatic organoid, liver organoid, heart organoid, or intestinal organoid, contains an adipose-derived stem cell and/or an endothelial cell. In various embodiments of any aspect delineated herein, the pancreatic islet organoid, pancreatic organoid, liver organoid, heart organoid, or intestinal organoid is vascularized. 
     In another aspect, the invention provides a method of generating a pancreatic islet organoid, the method comprising culturing an induced pluripotent stem cell (iPSC)-derived beta-like cell in a medium comprising Wnt4 or Wnt5a protein. In an embodiment, the induced pluripotent stem cell (iPSC)-derived beta-like cell is cultured in a 3-dimensional matrix. In an embodiment of the foregoing aspect, the Wnt4 or Wnt5a protein is a recombinant human Wnt4 or Wnt5a protein. In a particular embodiment, the medium comprises recombinant human Wnt4 protein. In another particular embodiment, the medium comprises recombinant human Wnt5a protein. 
     In another aspect the invention provides a cell culture comprising a human iPSC-derived beta-like cell and Wnt4 or Wnt5a protein. In an embodiment, the human iPSC-derived beta-like cell is in a three-dimensional matrix comprising gellan gum. In an embodiment, the Wnt4 or Wnt5a protein is a recombinant human Wnt4 or Wnt5a protein. In a particular embodiment, the medium comprises recombinant human Wnt4 protein. In another particular embodiment, the medium comprises recombinant human Wnt5a protein. 
     In another aspect, the invention provides a pancreatic islet organoid comprising an iPSC-derived beta-like cell cultured in medium comprising Wnt4 or Wnt5a protein, wherein the organoid is vascularized and exhibits glucose-stimulated insulin secretion (GSIS). In an embodiment, the organoid further exhibits KCl-stimulated insulin secretion or glucose stimulated insulin secretion. In an embodiment, the pancreatic islet organoid expresses Fltp and Esrrg genes. In an embodiment, the Wnt4 or Wnt5a protein is a recombinant human Wnt4 or Wnt5a protein. In a particular embodiment, the medium comprises recombinant human Wnt4 protein. In another particular embodiment, the medium comprises recombinant human Wnt5a protein. 
     In another aspect, the invention provides a non-human organism transplanted with the organoid defined in the above described aspects. 
     In another aspect, the invention provides a method of enhancing self organization of adipose-derived stem cells (ADSCs) for generating an induced pluripotent stem cell (iPSC)-derived organoid, the method comprising culturing the ADSCs in a 3-dimensional (3-D) culture matrix medium comprising a Wnt5a protein. In an embodiment of the method, the ADSCs are cultured in a 3-D culture matrix comprising gellan gum. In an embodiment, the ADSCs are cultured in the 3-D culture matrix medium comprising a Wnt5 protein and an iPSC-derived cell selected from an iPSC-derived beta-like cell, an iPSC-derived exocrine component cell, an iPSC-derived hepatocyte, an iPSC-derived cardiomyocyte, or an iPSC-derived intestinal cell. In an embodiment of the method, the iPSC-derived organoid is selected from a pancreatic islet organoid, pancreatic organoid, a liver organoid, a heart organoid, or an intestinal organoid. In an embodiment of the method, the induced pluripotent stem cell (iPSC)-derived organoid is a human induced pluripotent stem cell (hiPSC)-derived organoid. In an embodiment of the method, the Wnt5a protein is a recombinant human Wnt5a protein. In an embodiment of the method, the pancreatic islet organoid, pancreatic organoid, liver organoid, heart organoid, or intestinal organoid is derived from an iPSC-derived cell selected from an iPSC-derived beta-like cell, an iPSC-derived exocrine component cell, an iPSC-derived hepatocyte, an iPSC-derived cardiomyocyte, or an iPSC-derived intestinal cell, respectively. In an embodiment, of any of the above, the iPSC-derived cell is human. In another aspect, the invention provides a method of enhancing self organization of adipose-derived stem cells (ADSCs) for generating a pancreatic islet or pancreatic organoid, comprising culturing ADSCs in medium comprising Wnt5a protein. In an embodiment, the ADSCs are cultured in a 3-dimensional matrix comprising gellan gum. In another embodiment, the Wnt5a protein a recombinant human Wnt5a protein. In another aspect, the invention provides a pancreatic islet organoid, pancreatic organoid, a liver organoid, a heart organoid, or intestinal organoid produced by any of the above-delineated methods and embodiments thereof. 
     Compositions and articles defined by the invention were isolated or otherwise manufactured in connection with the examples provided below. Other features and advantages of the invention will be apparent from the detailed description, and from the claims. 
     Definitions 
     Unless defined otherwise, all technical and scientific terms used herein have the meaning commonly understood by a person skilled in the art to which this invention belongs. The following references provide one of skill with a general definition of many of the terms used in this invention: Singleton et al., Dictionary of Microbiology and Molecular Biology (2nd ed. 1994); The Cambridge Dictionary of Science and Technology (Walker ed., 1988); The Glossary of Genetics, 5th Ed., R. Rieger et al. (eds.), Springer Verlag (1991); and Hale &amp; Marham, The Harper Collins Dictionary of Biology (1991). As used herein, the following terms have the meanings ascribed to them below, unless specified otherwise. 
     By “AFP polypeptide” or “alpha-fetoprotein” is meant a protein or fragment thereof having at least 85% amino acid sequence identity to the sequence provided at NCBI Accession No. NP_001125.1 and having a biological activity of an AFP polypeptide. Exemplary biological activities of an AFP polypeptide include binding to copper, nickel, fatty acids, and bilirubin. The amino acid sequence provided at NCBI Accession No. NP_001125.1 is shown below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 1) 
                   
               
            
           
           
               
               
               
            
               
                   1 
                 mkwvesifli fllnftesrt lhrneygias ildsyqctae isladlatif faqfvqeaty 
                   
               
               
                   
               
               
                  61 
                 kevskmvkda ltaiekptgd eqssgclenq lpafleelch ekeilekygh sdccsqseeg 
               
               
                   
               
               
                 121 
                 rhncflahkk ptpasiplfq vpepvtscea yeedretfmn kfiyeiarrh pflyaptill 
               
               
                   
               
               
                 181 
                 waarydkiip scckaenave cfqtkaatvt kelresslln qhacavmknf gtrtfqaitv 
               
               
                   
               
               
                 241 
                 tklsqkftkv nfteiqklvl dvahvhehcc rgdvldclqd gekimsyics qqdtlsnkit 
               
               
                   
               
               
                 301 
                 eccklttler gqciihaend ekpeglspnl nrflgdrdfn qfssgeknif lasfvheysr 
               
               
                   
               
               
                 361 
                 rhpqlaysvi lrvakgygel lekcfqtenp lecqdkgeee lqkyiqesqa lakrscglfq 
               
               
                   
               
               
                 421 
                 klgeyylqna flvaytkkap qltsselmai trkmaataat ccqlsedkll acgegaadii 
               
               
                   
               
               
                 481 
                 ighlcirhem tpvnpgvgqc ctssyanrrp cfsslvvdet yvppafsddk fifhkdlcqa 
               
               
                   
               
               
                 541 
                 qgvalgtmkg eflinlvkqk pqiteeqlea viadfsglle kccqgqeqev cfaeegqkli 
               
               
                   
               
               
                 601 
                 sktraalgv 
               
            
           
         
       
     
     By “AFP polynucleotide” is meant a polynucleotide encoding a AFP polypeptide or fragment thereof. An exemplary AFP polynucleotide sequence is provided at NCBI Ref: NM_001134.2. The sequence provided at NCBI Ref: NM_001134.2 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 2) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 atattgtgct tccaccactg ccaataacaa aataactagc aaccatgaag tgggtggaat 
                   
               
               
                   
               
               
                 61 
                 caattttttt aattttccta ctaaatttta ctgaatccag aacactgcat agaaatgaat 
               
               
                   
               
               
                 121 
                 atggaatagc ttccatattg gattcttacc aatgtactgc agagataagt ttagctgacc 
               
               
                   
               
               
                 181 
                 tggctaccat attttttgcc cagtttgttc aagaagccac ttacaaggaa gtaagcaaaa 
               
               
                   
               
               
                 241 
                 tggtgaaaga tgcattgact gcaattgaga aacccactgg agatgaacag tcttcagggt 
               
               
                   
               
               
                 301 
                 gtttagaaaa ccagctacct gcctttctgg aagaactttg ccatgagaaa gaaattttgg 
               
               
                   
               
               
                 361 
                 agaagtacgg acattcagac tgctgcagcc aaagtgaaga gggaagacat aactgttttc 
               
               
                   
               
               
                 421 
                 ttgcacacaa aaagcccact ccagcatcga tcccactttt ccaagttcca gaacctgtca 
               
               
                   
               
               
                 481 
                 caagctgtga agcatatgaa gaagacaggg agacattcat gaacaaattc atttatgaga 
               
               
                   
               
               
                 541 
                 tagcaagaag gcatcccttc ctgtatgcac ctacaattct tctttgggct gctcgctatg 
               
               
                   
               
               
                 601 
                 acaaaataat tccatcttgc tgcaaagctg aaaatgcagt tgaatgcttc caaacaaagg 
               
               
                   
               
               
                 661 
                 cagcaacagt tacaaaagaa ttaagagaaa gcagcttgtt aaatcaacat gcatgtgcag 
               
               
                   
               
               
                 721 
                 taatgaaaaa ttttgggacc cgaactttcc aagccataac tgttactaaa ctgagtcaga 
               
               
                   
               
               
                 781 
                 agtttaccaa agttaatttt actgaaatcc agaaactagt cctggatgtg gcccatgtac 
               
               
                   
               
               
                 841 
                 atgagcactg ttgcagagga gatgtgctgg attgtctgca ggatggggaa aaaatcatgt 
               
               
                   
               
               
                 901 
                 cctacatatg ttctcaacaa gacactctgt caaacaaaat aacagaatgc tgcaaactga 
               
               
                   
               
               
                 961 
                 ccacgctgga acgtggtcaa tgtataattc atgcagaaaa tgatgaaaaa cctgaaggtc 
               
               
                   
               
               
                 1021 
                 tatctccaaa tctaaacagg tttttaggag atagagattt taaccaattt tcttcagggg 
               
               
                   
               
               
                 1081 
                 aaaaaaatat cttcttggca agttttgttc atgaatattc aagaagacat cctcagcttg 
               
               
                   
               
               
                 1141 
                 ctgtctcagt aattctaaga gttgctaaag gataccagga gttattggag aagtgtttcc 
               
               
                   
               
               
                 1201 
                 agactgaaaa ccctcttgaa tgccaagata aaggagaaga agaattacag aaatacatcc 
               
               
                   
               
               
                 1261 
                 aggagagcca agcattggca aagcgaagct gcggcctctt ccagaaacta ggagaatatt 
               
               
                   
               
               
                 1321 
                 acttacaaaa tgcgtttctc gttgcttaca caaagaaagc cccccagctg acctcgtcgg 
               
               
                   
               
               
                 1381 
                 agctgatggc catcaccaga aaaatggcag ccacagcagc cacttgttgc caactcagtg 
               
               
                   
               
               
                 1441 
                 aggacaaact attggcctgt ggcgagggag cggctgacat tattatcgga cacttatgta 
               
               
                   
               
               
                 1501 
                 tcagacatga aatgactcca gtaaaccctg gtgttggcca gtgctgcact tcttcatatg 
               
               
                   
               
               
                 1561 
                 ccaacaggag gccatgcttc agcagcttgg tggtggatga aacatatgtc cctcctgcat 
               
               
                   
               
               
                 1621 
                 tctctgatga caagttcatt ttccataagg atctgtgcca agctcagggt gtagcgctgc 
               
               
                   
               
               
                 1681 
                 aaacgatgaa gcaagagttt ctcattaacc ttgtgaagca aaagccacaa ataacagagg 
               
               
                   
               
               
                 1741 
                 aacaacttga ggctgtcatt gcagatttct caggcctgtt ggagaaatgc tgccaaggcc 
               
               
                   
               
               
                 1801 
                 aggaacagga agtctgcttt gctgaagagg gacaaaaact gatttcaaaa actcgtgctg 
               
               
                   
               
               
                 1861 
                 ctttgggagt ttaaattact tcaggggaag agaagacaaa acgagtcttt cattcggtgt 
               
               
                   
               
               
                 1921 
                 gaacttttct ctttaatttt aactgattta acactttttg tgaattaatg aaatgataaa 
               
               
                   
               
               
                 1981 
                 gacttttatg tgagatttcc ttatcacaga aataaaatat ctccaaatgt ttccttttca 
               
               
                   
               
               
                 2041 
                 aaaaaaaaaa aaaaaaa 
               
            
           
         
       
     
     By “ALB polypeptide” or “albumin” is meant a protein or fragment thereof having at least 85% amino acid sequence identity to the sequence provided at NCBI Accession No. NP_000468.1 and having a biological activity of ALB polypeptide. Exemplary biological activities of ALB polypeptide include binding to fatty acids, calcium ions, sodium ions, potassium ions, hormones, and bilirubin; stabilization of extracellular fluid volume; and, transport of plasma zinc. The amino acid sequence provided at NCBI Accession No. NP_000468.1 is shown below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 3) 
                   
               
            
           
           
               
               
               
            
               
                   1 
                 mkwvtfisll flfssaysrg vfrrdahkse vahrfkdlge enfkalvlia faqylqqcpf 
                   
               
               
                   
               
               
                  61 
                 edhvklvnev tefaktcvad esaencdksl htlfgdklct vatlretyge madccakqep 
               
               
                   
               
               
                 121 
                 ernecflqhk ddnpnlprlv rpevdvmcta fhdneetflk kylyeiarrh pyfyapellf 
               
               
                   
               
               
                 181 
                 fakrykaaft eccqaadkaa cllpkldelr degkassakq rlkcaslqkf gerafkawav 
               
               
                   
               
               
                 241 
                 arlsqrfpka efaevsklvt dltkvhtecc hgdllecadd radlakyice nqdsissklk 
               
               
                   
               
               
                 301 
                 eccekpllek shciaevend empadlpsla adfveskdvc knyaeakdvf lgmflyeyar 
               
               
                   
               
               
                 361 
                 rhpdysvvll lrlaktyett lekccaaadp hecyakvfde fkplveepqn likqncelfe 
               
               
                   
               
               
                 421 
                 qlgeykfqna llvrytkkvp qvstptlvev srnlgkvgsk cckhpeakrm pcaedylsvv 
               
               
                   
               
               
                 481 
                 lnqlcvlhek tpvsdrvtkc cteslvnrrp cfsalevdet yvpkefnaet ftfhadictl 
               
               
                   
               
               
                 541 
                 sekergikkg talvelvkhk pkatkeqlka vmddfaafve kcckaddket cfaeegkklv 
               
               
                   
               
               
                 601 
                 aasqaalgl 
               
            
           
         
       
     
     By “ALB polynucleotide” is meant a polynucleotide encoding a ALB polypeptide or fragment thereof. An exemplary AFP polynucleotide sequence is provided at NCBI Ref: NM_000477.5. The sequence provided at NCBI Ref: NM_000477.5 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 4) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 agtatattag tgctaatttc cctccgtttg tcctagcttt tctcttctgt caaccccaca 
                   
               
               
                   
               
               
                 61 
                 cgcctttggc acaatgaagt gggtaacctt tatttccctt ctttttctct ttagctcggc 
               
               
                   
               
               
                 121 
                 ttattccagg ggtgtgtttc gtcgagatgc acacaagagt gaggttgctc atcggtttaa 
               
               
                   
               
               
                 181 
                 agatttggga gaagaaaatt tcaaagcctt ggtgttgatt gcctttgctc agtatcttca 
               
               
                   
               
               
                 241 
                 gcagtgtcca tttgaagatc atgtaaaatt agtgaatgaa gtaactgaat ttgcaaaaac 
               
               
                   
               
               
                 301 
                 atgtgttgct gatgagtcag ctgaaaattg tgacaaatca cttcataccc tttttggaga 
               
               
                   
               
               
                 361 
                 caaattatgc acagttgcaa ctcttcgtga aacctatggt gaaatggctg actgctgtgc 
               
               
                   
               
               
                 421 
                 aaaacaagaa cctgagagaa atgaatgctt cttgcaacac aaagatgaca acccaaacct 
               
               
                   
               
               
                 481 
                 cccccgattg gtgagaccag aggttgatgt gatgtgcact gcttttcatg acaatgaaga 
               
               
                   
               
               
                 541 
                 gacatttttg aaaaaatact tatatgaaat tgccagaaga catccttact tttatgcccc 
               
               
                   
               
               
                 601 
                 ggaactcctt ttctttgcta aaaggtataa agctgctttt acagaatgtt gccaagctgc 
               
               
                   
               
               
                 661 
                 tgataaagct gcctgcctgt tgccaaagct cgatgaactt cgggatgaag ggaaggcttc 
               
               
                   
               
               
                 721 
                 gtctgccaaa cagagactca agtgtgccag tctccaaaaa tttggagaaa gagctttcaa 
               
               
                   
               
               
                 781 
                 agcatgggca gtagctcgcc tgagccagag atttcccaaa gctgagtttg cagaagtttc 
               
               
                   
               
               
                 841 
                 caagttagtg acagatctta ccaaagtcca cacggaatgc tgccatggag atctgcttga 
               
               
                   
               
               
                 901 
                 atgtgctgat gacagggcgg accttgccaa gtatatctgt gaaaatcaag attcgatctc 
               
               
                   
               
               
                 961 
                 cagtaaactg aaggaatgct gtgaaaaacc tctgttggaa aaatcccact gcattgccga 
               
               
                   
               
               
                 1021 
                 agtggaaaat gatgagatgc ctgctgactt gccttcatta gctgctgatt ttgttgaaag 
               
               
                   
               
               
                 1081 
                 taaggatgtt tgcaaaaact atgctgaggc aaaggatgtc ttcctgggca tgtttttgta 
               
               
                   
               
               
                 1141 
                 tgaatatgca agaaggcatc ctgattactc tgtcgtgctg ctgctgagac ttgccaagac 
               
               
                   
               
               
                 1201 
                 atatgaaacc actctagaga agtgctgtgc cgctgcagat cctcatgaat gctatgccaa 
               
               
                   
               
               
                 1261 
                 agtgttcgat gaatttaaac ctcttgtgga agagcctcag aatttaatca aacaaaattg 
               
               
                   
               
               
                 1321 
                 tgagcttttt gagcagcttg gagagtacaa attccagaat gcgctattag ttcgttacac 
               
               
                   
               
               
                 1381 
                 caagaaagta ccccaagtgt caactccaac tcttgtagag gtctcaagaa acctaggaaa 
               
               
                   
               
               
                 1441 
                 agtgggcagc aaatgttgta aacatcctga agcaaaaaga atgccctgtg cagaagacta 
               
               
                   
               
               
                 1501 
                 tctatccgtg gtcctgaacc agttatgtgt gttgcatgag aaaacgccag taagtgacag 
               
               
                   
               
               
                 1561 
                 agtcaccaaa tgctgcacag aatccttggt gaacaggcga ccatgctttt cagctctgga 
               
               
                   
               
               
                 1621 
                 agtcgatgaa acatacgttc ccaaagagtt taatgctgaa acattcacct tccatgcaga 
               
               
                   
               
               
                 1681 
                 tatatgcaca ctttctgaga aggagagaca aatcaagaaa caaactgcac ttgttgagct 
               
               
                   
               
               
                 1741 
                 cgtgaaacac aagcccaagg caacaaaaga gcaactgaaa gctgttatgg atgatttcgc 
               
               
                   
               
               
                 1801 
                 agcttttgta gagaagtgct gcaaggctga cgataaggag acctgctttg ccgaggaggg 
               
               
                   
               
               
                 1861 
                 taaaaaactt gttgctgcaa gtcaagctgc cttaggctta taacatcaca tttaaaagca 
               
               
                   
               
               
                 1921 
                 tctcagccta ccatgagaat aagagaaaga aaatgaagat caaaagctta ttcatctgtt 
               
               
                   
               
               
                 1981 
                 tttctttttc gttggtgtaa agccaacacc ctgtctaaaa aacataaatt tctttaatca 
               
               
                   
               
               
                 2041 
                 ttttgcctct tttctctgtg cttcaattaa taaaaaatgg aaagaatcta atagagtggt 
               
               
                   
               
               
                 2101 
                 acagcactgt tatttttcaa agatgtgttg ctatcctgaa aattctgtag gttctgtgga 
               
               
                   
               
               
                 2161 
                 agttccagtg ttctctctta ttccacttcg gtagaggatt tctagtttct tgtgggctaa 
               
               
                   
               
               
                 2221 
                 ttaaataaat cattaatact cttctaaaaa aaaaaaaaaa aaaa 
               
            
           
         
       
     
     By “agent” is meant any small molecule chemical compound, antibody, nucleic acid molecule, or polypeptide, or fragments thereof. 
     By “ameliorate” is meant decrease, suppress, attenuate, diminish, arrest, or stabilize the development or progression of a disease. 
     By “altered” is meant an increase or decrease. An increase is any positive change, e.g., by at least about 5%, 10%, or 20%; by at least about 25%, 50%, 75%, or even by 100%, 200%, 300% or more. A decrease is a negative change, e.g., a decrease by at least about 5%, 10%, or 20%; by at least about 25%, 50%, 75%; or even an increase by 100%, 200%, 300% or more. 
     In this disclosure, “comprises,” “comprising,” “containing” and “having” and the like can have the meaning ascribed to them in U.S. Patent law and can mean “includes,” “including,” and the like; “consisting essentially of” or “consists essentially” likewise has the meaning ascribed in U.S. Patent law and the term is open-ended, allowing for the presence of more than that which is recited so long as basic or novel characteristics of that which is recited is not changed by the presence of more than that which is recited, but excludes prior art embodiments. 
     By “CDX2 polypeptide” is meant a protein or fragment thereof having at least 85% amino acid sequence identity to the sequence provided at NCBI Accession No. NP_001256.3 and having transcription factor activity. The amino acid sequence provided at NCBI Accession No. NP_001256.3 is shown below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 5) 
                   
               
            
           
           
               
               
               
            
               
                   1 
                 myvsylldkd vsmypssvrh sgglnlapqn fvsppqypdy ggyhvaaaaa aaanldsaqs 
                   
               
               
                   
               
               
                  61 
                 pgpswpaayg aplredwngy apggaaaaan avahglnggs paaamgyssp adyhphhhph 
               
               
                   
               
               
                 121 
                 hhphhpaaap scasgllgtl npgppgpaat aaaeqlspgg qrrnlcewmr kpaqqslgsq 
               
               
                   
               
               
                 181 
                 vktrtkdkyr vvytdhqrle lekefhysry itirrkaela atlglserqv kiwfqnrrak 
               
               
                   
               
               
                 241 
                 erkinkkklq qqqqqqppqp pppppqppqp qpgplrsvpe plspvsslqa sysgsvpgvl 
               
               
                   
               
               
                 301 
                 gptggvinpt vtq 
               
            
           
         
       
     
     By “CDX2 polynucleotide” is meant a polynucleotide encoding a CDX2 polypeptide or fragment thereof. An exemplary CDX2 polynucleotide sequence is provided at NCBI Ref: NM_001265.4. The sequence provided at NCBI Ref: NM_001265.4 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 6) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 ctccaaccat tggtgtctgt gtcattacta atagagtctt gtaaacactc gttaatcacg 
                   
               
               
                   
               
               
                 61 
                 gaaggccgcc ggcctggggc tccgcacgcc agcctgtggc gggtcttccc cgcctctgca 
               
               
                   
               
               
                 121 
                 gcctagtggg aaggaggtgg gaggaaagaa ggaagaaagg gagggaggga ggaggcaggc 
               
               
                   
               
               
                 181 
                 cagagggagg gaccgcctcg gaggcagaag agccgcgagg agccagcgga gcaccgcggg 
               
               
                   
               
               
                 241 
                 ctggggcgca gccacccgcc gctcctcgag tcccctcgcc cctttccctt cgtgcccccc 
               
               
                   
               
               
                 301 
                 ggcagcctcc agcgtcggtc cccaggcagc atggtgaggt ctgctcccgg accctcgcca 
               
               
                   
               
               
                 361 
                 ccatgtacgt gagctacctc ctggacaagg acgtgagcat gtaccctagc tccgtgcgcc 
               
               
                   
               
               
                 421 
                 actctggcgg cctcaacctg gcgccgcaga acttcgtcag ccccccgcag tacccggact 
               
               
                   
               
               
                 481 
                 acggcggtta ccacgtggcg gccgcagctg cagcggcagc gaacttggac agcgcgcagt 
               
               
                   
               
               
                 541 
                 ccccggggcc atcctggccg gcagcgtatg gcgccccact ccgggaggac tggaatggct 
               
               
                   
               
               
                 601 
                 acgcgcccgg aggcgccgcg gccgccgcca acgccgtggc tcacggcctc aacggtggct 
               
               
                   
               
               
                 661 
                 ccccggccgc agccatgggc tacagcagcc ccgcagacta ccatccgcac caccacccgc 
               
               
                   
               
               
                 721 
                 atcaccaccc gcaccacccg gccgccgcgc cttcctgcgc ttctgggctg ctgcaaacgc 
               
               
                   
               
               
                 781 
                 tcaaccccgg ccctcctggg cccgccgcca ccgctgccgc cgagcagctg tctcccggcg 
               
               
                   
               
               
                 841 
                 gccagcggcg gaacctgtgc gagtggatgc ggaagccggc gcagcagtcc ctcggcagcc 
               
               
                   
               
               
                 901 
                 aagtgaaaac caggacgaaa gacaaatatc gagtggtgta cacggaccac cagcggctgg 
               
               
                   
               
               
                 961 
                 agctggagaa ggagtttcac tacagtcgct acatcaccat ccggaggaaa gccgagctag 
               
               
                   
               
               
                 1021 
                 ccgccacgct ggggctctct gagaggcagg ttaaaatctg gtttcagaac cgcagagcaa 
               
               
                   
               
               
                 1081 
                 aggagaggaa aatcaacaag aagaagttgc agcagcaaca gcagcagcag ccaccacagc 
               
               
                   
               
               
                 1141 
                 cgcctccgcc gccaccacag cctccccagc ctcagccagg tcctctgaga agtgtcccag 
               
               
                   
               
               
                 1201 
                 agcccttgag tccggtgtct tccctgcaag cctcagtgtc tggctctgtc cctggggttc 
               
               
                   
               
               
                 1261 
                 tggggccaac tgggggggtg ctaaacccca ccgtcaccca gtgacccacc gggttctgca 
               
               
                   
               
               
                 1321 
                 gcggcagagc aattccaggc tgagccatga ggagcgtgga ctctgctaga ctcctcagga 
               
               
                   
               
               
                 1381 
                 gagacccctc ccctcccacc cacagccata gacctacaga cctggctctc agaggaaaaa 
               
               
                   
               
               
                 1441 
                 tgggagccag gagtaagaca agtgggattt ggggcctcaa gaaatatact ctcccagatt 
               
               
                   
               
               
                 1501 
                 tttacttttt cccatctggc tttttctgcc actgaggaga cagaaagcct ccgctgggct 
               
               
                   
               
               
                 1561 
                 tcattccgga ctggcagaag cattgcctgg actgaccaca ccaaccaggc cttcatcctc 
               
               
                   
               
               
                 1621 
                 ctccccagct cttctcttcc tagatctgca ggctgcacct ctggctagag ccgaggggag 
               
               
                   
               
               
                 1681 
                 agagggactc aagggaaagg caagcttgag gccaagatgg ctgctgcctg ctcatggccc 
               
               
                   
               
               
                 1741 
                 tcggaggtcc agctgggcct cctgcctccg ggcaggcaag gtttacactg cggaagccaa 
               
               
                   
               
               
                 1801 
                 aggcagctaa gatagaaagc tggactgacc aaagactgca gaacccccag gtggcctgcg 
               
               
                   
               
               
                 1861 
                 tcttttttct cttcccttcc cagaccagga aaggcttggc tggtgtatgc acagggtgtg 
               
               
                   
               
               
                 1921 
                 gtatgagggg gtggttattg gactccaggc ctgaccaggg ggcccgaaca gggacttgtt 
               
               
                   
               
               
                 1981 
                 tagagagcct gtcaccagag cttctctggg ctgaatgtat gtcagtgcta taaatgccag 
               
               
                   
               
               
                 2041 
                 agccaacctg gacttcctgt cattttcaca atcttggggc tgatgaagaa gggggtgggg 
               
               
                   
               
               
                 2101 
                 ggagtttgtg ttgttgttgc tgctgtttgg gttgttggtc tgtgtaacat ccaagccaga 
               
               
                   
               
               
                 2161 
                 gtttttaaag ccttctggat ccatgggggg agaagtgata tggtgaaggg aagtggggag 
               
               
                   
               
               
                 2221 
                 tatttgaaca cagttgaatt ttttctaaaa agaaaaagag ataaatgagc tttccagatt 
               
               
                   
               
               
                 2281 
                 tcagattctg tatttatctt cagattttgt ctgcaactat tttttatttt ttaaagaaat 
               
               
                   
               
               
                 2341 
                 gaaatatctt caaaaaaaaa aaaaaaaaaa 
               
            
           
         
       
     
     By “CYP3A7 polypeptide” or “cytochrome P450” is meant a protein or fragment thereof having at least 85% amino acid sequence identity to the sequence provided at NCBI Accession No. NP_000756.3 and having monooxygenase activity. The amino acid sequence provided at NCBI Accession No. NP_000756.3 is shown below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 7) 
                   
               
            
           
           
               
               
               
            
               
                   1 
                 mdlipnlave twlllavsli llylygtrth glfkklgipg ptplpflgna lsfrkgywtf 
                   
               
               
                   
               
               
                  61 
                 dmecykkyrk vwgiydcqqp mlaitdpdmi ktvlvkecys vftnrrpfgp vgfmknaisi 
               
               
                   
               
               
                 121 
                 aedeewkrir sllsptftsg klkemvpiia qygdvlvrnl rreaetgkpv tlkhvfgays 
               
               
                   
               
               
                 181 
                 mdvitstsfg vsidslnnpq dpfventkkl lrfnpldpfv lsikvfpflt pilealnitv 
               
               
                   
               
               
                 241 
                 fprkvisflt ksvkqikegr lketqkhrvd flqlmidsqn skdsethkal sdlelmaqsi 
               
               
                   
               
               
                 301 
                 ififagyett ssvlsfiiye lathpdvqqk vqkeidtvlp nkapptydtv lqleyldmvv 
               
               
                   
               
               
                 361 
                 netlrlfpva mrlervckkd veingmfipk gvvvmipsyv lhhdpkywte pekflperfs 
               
               
                   
               
               
                 421 
                 kknkdnidpy iytpfgsgpr ncigmrfalv nmklalvrvl qnfsfkpcke tqiplklrfg 
               
               
                   
               
               
                 481 
                 gllltekpiv lkaesrdetv sga 
               
            
           
         
       
     
     By “CYP3A7 polynucleotide” is meant a polynucleotide encoding a CYP3A7 polypeptide or fragment thereof. An exemplary AFP polynucleotide sequence is provided at NCBI Ref: NM_000765.4. The sequence provided at NCBI Ref: NM_000765.4 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 8) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 aatcactgct gtgcagggca ggaaagctcc acacacacag cccagcaaac agcagcacgc 
                   
               
               
                   
               
               
                 61 
                 tgctgaaaaa aagactcaga ggagagagat aaggaaggaa agtagtgatg gatctcatcc 
               
               
                   
               
               
                 121 
                 caaacttggc cgtggaaacc tggcttctcc tggctgtcag cctgatactc ctctatctat 
               
               
                   
               
               
                 181 
                 atggaacccg tacacatgga ctttttaaga agcttggaat tccagggccc acacctctgc 
               
               
                   
               
               
                 241 
                 cttttttggg aaatgctttg tccttccgta agggctattg gacgtttgac atggaatgtt 
               
               
                   
               
               
                 301 
                 ataaaaagta tagaaaagtc tggggtattt atgactgtca acagcctatg ctggctatca 
               
               
                   
               
               
                 361 
                 cagatcccga catgatcaaa acagtgctag tgaaagaatg ttattctgtc ttcacaaacc 
               
               
                   
               
               
                 421 
                 ggaggccttt cgggccagtg ggatttatga aaaatgccat ctctatagct gaggatgaag 
               
               
                   
               
               
                 481 
                 aatggaagag aatacgatca ttgctgtctc caacattcac cagcggaaaa ctcaaggaga 
               
               
                   
               
               
                 541 
                 tggtccctat cattgcccag tatggagatg tgttggtgag aaatctgagg cgggaagcag 
               
               
                   
               
               
                 601 
                 agacaggcaa gcctgtcacc ttgaaacacg tctttggggc ctacagcatg gatgtgatca 
               
               
                   
               
               
                 661 
                 ctagcacatc atttggagtg agcatcgact ctctcaacaa tccacaagac ccctttgtgg 
               
               
                   
               
               
                 721 
                 aaaacaccaa gaagctttta agatttaatc cattagatcc attcgttctc tcaataaaag 
               
               
                   
               
               
                 781 
                 tctttccatt ccttacccca attcttgaag cattaaatat cactgtgttt ccaagaaaag 
               
               
                   
               
               
                 841 
                 ttataagttt tctaacaaaa tctgtaaaac agataaaaga aggtcgcctc aaagagacac 
               
               
                   
               
               
                 901 
                 aaaagcaccg agtggatttc cttcagctga tgattgactc tcagaattca aaagactctg 
               
               
                   
               
               
                 961 
                 agacccacaa agctctgtct gatctggagc tcatggccca atcaattatc tttatttttg 
               
               
                   
               
               
                 1021 
                 ctggctatga aaccacgagc agtgttctct ccttcattat atatgaactg gccactcacc 
               
               
                   
               
               
                 1081 
                 ctgatgtcca gcagaaagtg cagaaggaaa ttgatacagt tttacccaat aaggcaccac 
               
               
                   
               
               
                 1141 
                 ccacctatga tactgtgcta cagttggagt atcttgacat ggtggtgaat gaaacactca 
               
               
                   
               
               
                 1201 
                 gattattccc agttgctatg agacttgaga gggtctgcaa aaaagatgtt gaaatcaatg 
               
               
                   
               
               
                 1261 
                 ggatgtttat tcccaaaggg gtggtggtga tgattccaag ctatgttctt catcatgacc 
               
               
                   
               
               
                 1321 
                 caaagtactg gacagagcct gagaagttcc tccctgaaag gttcagtaaa aagaacaagg 
               
               
                   
               
               
                 1381 
                 acaacataga tccttacata tacacaccct ttggaagtgg acccagaaac tgcattggca 
               
               
                   
               
               
                 1441 
                 tgaggtttgc tctcgtgaac atgaaacttg ctctagtcag agtccttcag aacttctcct 
               
               
                   
               
               
                 1501 
                 tcaaaccttg taaagaaaca cagatccccc tgaaattacg ctttggagga cttcttctaa 
               
               
                   
               
               
                 1561 
                 cagaaaaacc cattgttcta aaggctgagt caagggatga gaccgtaagt ggagcctgat 
               
               
                   
               
               
                 1621 
                 ttccctaagg acttctggtt tgctctttaa gaaagctgtg ccccagaaca ccagagacct 
               
               
                   
               
               
                 1681 
                 caaattactt tacaaataga accctgaaat gaagacgggc ttcatccaat gtgctgcata 
               
               
                   
               
               
                 1741 
                 aataatcagg gattctgtac gtgcattgtg ctctctcatg gtctgtatag agtgttatac 
               
               
                   
               
               
                 1801 
                 ttggtaatat agaggagatg accaaatcag tgctggggaa gtagatttgg cttctctgct 
               
               
                   
               
               
                 1861 
                 tctcatagga ctatctccac cacccccagt tagcaccatt aactcctcct gagctctgat 
               
               
                   
               
               
                 1921 
                 aacataatta acatttctca ataatttcaa ccacaatcat taataaaaat aggaattatt 
               
               
                   
               
               
                 1981 
                 ttgatggctc taacagtgac atttatatca tgtgttatat ctgtagtatt ctatagtaag 
               
               
                   
               
               
                 2041 
                 ctttatatta agcaaatcaa taaaaacctc tttacaaaag taaaaaaaaa aaaaaaaaa 
               
            
           
         
       
     
     “Detect” refers to identifying the presence, absence or amount of the analyte to be detected. 
     By “detectable label” is meant a composition that when linked to a molecule of interest renders the latter detectable, via spectroscopic, photochemical, biochemical, immunochemical, or chemical means. For example, useful labels include radioactive isotopes, magnetic beads, metallic beads, colloidal particles, fluorescent dyes, electron-dense reagents, enzymes (for example, as commonly used in an ELISA), biotin, digoxigenin, or haptens. 
     “Differentiation” refers to the developmental process of lineage commitment. Differentiation can be assayed by measuring an increase in one or more cell specific markers relative to their expression in a corresponding undifferentiated control cell. A “lineage” refers to a pathway of cellular development, in which precursor or “progenitor” cells undergo progressive physiological changes to become a specified cell type having a characteristic function. In some embodiments, the cell type is a beta cell. In some embodiments, the cell type is an alpha cell, delta cell, or duct cell. In some other embodiments, the cell type is a hepatocyte. In still other embodiments, the cell type is a cardiomyocyte. In some embodiments, the cell type is a intestinal cell. Differentiation occurs in stages, whereby cells gradually become more specified until they reach full maturity, which is also referred to as “terminal differentiation.” A “terminally differentiated cell” is a cell that has committed to a specific lineage, and has reached the end stage of differentiation (i.e., a cell that has fully matured). In some embodiments, an induced pluripotent stem cell (iPSC) is differentiated into a beta-like cell, an alpha-like cell, a delta-like cell, or a duct-like cell. In some other embodiments, an induced pluripotent stem cell (iPSC) is differentiated into a hepatocyte, cardiomyocyte, or intestinal cell. 
     A “de-differentiated cell” is a cell in which the process of differentiation has been, at least to some degree, reversed. De-differentiation can be assayed, for example, by identifying a reduction in the expression of one or more cell specific markers relative to their expression in a corresponding control cell. Alternatively, de-differentiation can be assayed by measuring an increase in one or more markers typically expressed in an embryonic stem cell, a pluripotent or multi-potent cell type, or expressed at an earlier stage of development. In some embodiments, the de-differentiated cell is an induced pluripotent stem cell (iPSC). In certain embodiments, the de-differentiated cell is a human induced pluripotent stem cell (iPSC). 
     By “disease” is meant any condition or disorder that damages or interferes with the normal function of a cell, tissue, or organ. Examples of diseases include type 1 diabetes, type 2 diabetes, and pancreatic cancer. 
     By “effective amount” is meant the amount of a therapeutic agent or organoid required to ameliorate the symptoms of a disease in a subject relative to an untreated subject. The effective amount of a therapeutic used to practice the present invention for therapeutic treatment of a disease varies depending upon the manner of administration, the age, body weight, and general health of the subject. Ultimately, the attending physician or veterinarian will decide the appropriate amount and dosage regimen. Such amount is referred to as an “effective” amount. In some embodiments, the therapeutic organoid is a pancreatic islet organoid. In some other embodiments, an effective amount of a pancreatic islet organoid is administered to a subject having type 1 or type 2 diabetes. 
     By “ESRRG polypeptide” is meant a protein or fragment thereof having at least 85% amino acid sequence identity to the sequence provided at NCBI Accession No. NP_001230448.1 and having nuclear hormone receptor activity. The amino acid sequence provided at NCBI Accession No. NP_001230448.1 is shown below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 9) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 msnkdrhids scssfiktep sspasltdsv nhhspggssd asgsysstmn ghqngldspp 
                   
               
               
                   
               
               
                 61 
                 lypsapilgg sgpvrklydd csstivedpq tkceymlnsm pkrlclvcgd iasgyhygva 
               
               
                   
               
               
                 121 
                 sceackaffk rtiqgnieys cpatneceit krrrkscqac rfmkclkvgm lkegvrldrv 
               
               
                   
               
               
                 181 
                 rggrqkykrr idaenspyln pqlvqpakkp ynkivshllv aepekiyamp dptvpdsdik 
               
               
                   
               
               
                 241 
                 altticdlad relvviigwa khipgfstls ladqmsllqs awmeililgv vyrslsfede 
               
               
                   
               
               
                 301 
                 lvyaddyimd edqsklagll dlnnailqlv kkyksmklek eefvtlkaia lansdsmhie 
               
               
                   
               
               
                 361 
                 dveavqklqd vlhealqdye agqhmedprr agkmlmtlpl lrgtstkavq hfyniklegk 
               
               
                   
               
               
                 421 
                 vpmhklflem leakv 
               
            
           
         
       
     
     By “ESRRG polynucleotide” is meant a polynucleotide encoding a ESRRG polypeptide or fragment thereof. An exemplary ESRRG polynucleotide sequence is provided at NCBI Ref: NM_001243519.1. The sequence provided at NCBI Ref: NM_001243519.1 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 10) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 aagctccaat cggggcttta agtccttgat taggagagtg tgagagcttt ggtcccaact 
                   
               
               
                   
               
               
                 61 
                 ggctgtgcct ataggcttgt cactaggaga acatttgtgt taattgcact gtgctctgtc 
               
               
                   
               
               
                 121 
                 aaggaaactt tgatttatag ctggggtgca caaataatgg ttgccggtcg cacatggatt 
               
               
                   
               
               
                 181 
                 cggtagaact ttgccttcct gaatcttttt ccctgcacta cgaggaagag tagacttgaa 
               
               
                   
               
               
                 241 
                 tgagacctgc ctcatcagtc atgggatcat agtgtcacag atggaaaagc aactatcagc 
               
               
                   
               
               
                 301 
                 tgaattgtac tgaactacac acttggctaa ttcatcttat tgctctacac atctaaagga 
               
               
                   
               
               
                 361 
                 aggctcattc tgttcttgga gtctagacag catcaggagt tgggctcagt gaacaaaact 
               
               
                   
               
               
                 421 
                 ttaatgtcta gagcatttat gagggtttta atgattggaa aatctatcct gagaatgtgg 
               
               
                   
               
               
                 481 
                 tcaccatatg tgacagcctt gctttctatc ttgtcttcag tttctggggc ttctctgcag 
               
               
                   
               
               
                 541 
                 aatgtcaaac aaagatcgac acattgattc cagctgttcg tccttcatca agacggaacc 
               
               
                   
               
               
                 601 
                 ttccagccca gcctccctga cggacagcgt caaccaccac agccctggtg gctcttcaga 
               
               
                   
               
               
                 661 
                 cgccagtggg agctacagtt caaccatgaa tggccatcag aacggacttg actcgccacc 
               
               
                   
               
               
                 721 
                 tctctaccct tctgctccta tcctgggagg tagtgggcct gtcaggaaac tgtatgatga 
               
               
                   
               
               
                 781 
                 ctgctccagc accattgttg aagatcccca gaccaagtgt gaatacatgc tcaactcgat 
               
               
                   
               
               
                 841 
                 gcccaagaga ctgtgtttag tgtgtggtga catcgcttct gggtaccact atggggtagc 
               
               
                   
               
               
                 901 
                 atcatgtgaa gcctgcaagg cattcttcaa gaggacaatt caaggcaata tagaatacag 
               
               
                   
               
               
                 961 
                 ctgccctgcc acgaatgaat gtgaaatcac aaagcgcaga cgtaaatcct gccaggcttg 
               
               
                   
               
               
                 1021 
                 ccgcttcatg aagtgtttaa aagtgggcat gctgaaagaa ggggtgcgtc ttgacagagt 
               
               
                   
               
               
                 1081 
                 acgtggaggt cggcagaagt acaagcgcag gatagatgcg gagaacagcc catacctgaa 
               
               
                   
               
               
                 1141 
                 ccctcagctg gttcagccag ccaaaaagcc atataacaag attgtctcac atttgttggt 
               
               
                   
               
               
                 1201 
                 ggctgaaccg gagaagatct atgccatgcc tgaccctact gtccccgaca gtgacatcaa 
               
               
                   
               
               
                 1261 
                 agccctcact acactgtgtg acttggccga ccgagagttg gtggttatca ttggatgggc 
               
               
                   
               
               
                 1321 
                 gaagcatatt ccaggcttct ccacgctgtc cctggcggac cagatgagcc ttctgcagag 
               
               
                   
               
               
                 1381 
                 tgcttggatg gaaattttga tccttggtgt cgtataccgg tctctttcgt ttgaggatga 
               
               
                   
               
               
                 1441 
                 acttgtctat gcagacgatt atataatgga cgaagaccag tccaaattag caggccttct 
               
               
                   
               
               
                 1501 
                 tgatctaaat aatgctatcc tgcagctggt aaagaaatac aagagcatga agctggaaaa 
               
               
                   
               
               
                 1561 
                 agaagaattt gtcaccctca aagctatagc tcttgctaat tcagactcca tgcacataga 
               
               
                   
               
               
                 1621 
                 agatgttgaa gccgttcaga agcttcagga tgtcttacat gaagcgctgc aggattatga 
               
               
                   
               
               
                 1681 
                 agctggccag cacatggaag accctcgtcg agctggcaag atgctgatga cactgccact 
               
               
                   
               
               
                 1741 
                 cctgaggcag acctctacca aggccgtgca gcatttctac aacatcaaac tagaaggcaa 
               
               
                   
               
               
                 1801 
                 agtcccaatg cacaaacttt ttttggaaat gttggaggcc aaggtctgac taaaagctcc 
               
               
                   
               
               
                 1861 
                 ctgggccttc ccatccttca tgttgaaaaa gggaaaataa acccaagagt gatgtcgaag 
               
               
                   
               
               
                 1921 
                 aaacttagag tttagttaac aacatcaaaa atcaacagac tgcactgata atttagcagc 
               
               
                   
               
               
                 1981 
                 aagactatga agcagctttc agattcctcc ataggttcct gatgagtttc tttctacttt 
               
               
                   
               
               
                 2041 
                 ctccatcatc ttctttcctc tttcttccca catttctctt tctctttatt ttttctcctt 
               
               
                   
               
               
                 2101 
                 ttcttctttc acctccctta tttctttgct tctttcattc ctagttccca ttctccttta 
               
               
                   
               
               
                 2161 
                 ttttcttccc gtctgcctgc cttctttctt ttctttacct actctcattc ctctcttttc 
               
               
                   
               
               
                 2221 
                 tcatccttcc ccttttttct aaatttgaaa tagctttagt ttaaaaaaaa atcctccctt 
               
               
                   
               
               
                 2281 
                 ccccctttcc tttccctttc tttccttttt ccctttcctt ttccctttcc tttcctttcc 
               
               
                   
               
               
                 2341 
                 tcttgacctt ctttccatct ttctttttct tccttctgct gctgaacttt taaaagaggt 
               
               
                   
               
               
                 2401 
                 ctctaactga agagagatgg aagccagccc tgccaaagga tggagatcca taatatggat 
               
               
                   
               
               
                 2461 
                 gccagtgaac ttattgtgaa ccatactgtc cccaatgact aaggaatcaa agagagagaa 
               
               
                   
               
               
                 2521 
                 ccaacgttcc taaaagtaca gtgcaacata tacaaattga ctgagtgcag tattagattt 
               
               
                   
               
               
                 2581 
                 catgggagca gcctctaatt agacaactta agcaacgttg catcggctgc ttcttatcat 
               
               
                   
               
               
                 2641 
                 tgcttttcca tctagatcag ttacagccat ttgattcctt aattgttttt tcaagtcttc 
               
               
                   
               
               
                 2701 
                 caggtatttg ttagtttagc tactatgtaa ctttttcagg gaatagttta agctttattc 
               
               
                   
               
               
                 2761 
                 attcatgcaa tactaaagag aaataagaat actgcaattt tgtgctggct ttgaacaatt 
               
               
                   
               
               
                 2821 
                 acgaacaata atgaaggaca aatgaatcct gaaggaagat ttttaaaaat gttttgtttc 
               
               
                   
               
               
                 2881 
                 ttcttacaaa tggagatttt tttgtaccag ctttaccact tttcagccat ttattaatat 
               
               
                   
               
               
                 2941 
                 gggaatttaa cttactcaag caatagttga agggaaggtg catattatca cggatgcaat 
               
               
                   
               
               
                 3001 
                 ttatgttgtg tgccagtctg gtcccaaaca tcaatttctt aacatgagct ccagtttacc 
               
               
                   
               
               
                 3061 
                 taaatgttca ctgacacaaa ggatgagatt acacctacag tgactctgag tagtcacata 
               
               
                   
               
               
                 3121 
                 tataagcact gcacatgaga tatagatccg tagaattgtc aggagtgcac ctctctactt 
               
               
                   
               
               
                 3181 
                 gggaggtaca attgccatat gatttctagc tgccatggtg gttaggaatg tgatactgcc 
               
               
                   
               
               
                 3241 
                 tgtttgcaaa gtcacagacc ttgcctcaga aggagctgtg agccagtatt catttaagag 
               
               
                   
               
               
                 3301 
                 gcaataaggc aaatgccaga attaaaaaaa aaaatcatca aagacagaaa atgcctgacc 
               
               
                   
               
               
                 3361 
                 aaattctaaa acctaatcca tataagttta ttcatttagg aatgttcgtt taaattaatc 
               
               
                   
               
               
                 3421 
                 tgcagttttt accaagagct aagccaatat atgtgctttt caaccagtat tgtcacagca 
               
               
                   
               
               
                 3481 
                 tgaaagtcaa gtcaggttcc agactgttaa gaggtgtaat ctaatgaaga aatcaattag 
               
               
                   
               
               
                 3541 
                 atgccccgaa atctacagtc gctgaataac caataaacag taacctccat caaatgctat 
               
               
                   
               
               
                 3601 
                 accaatggac cagtgttagt agctgctccc tgtattatgt gaacagtctt attctatgta 
               
               
                   
               
               
                 3661 
                 cacagatgta attaaaattg taatcctaac aaacaaaaga aatgtagttc agcttttcaa 
               
               
                   
               
               
                 3721 
                 tgtttcatgt ttgctgtgct tttctgaatt ttatgttgca ttcaaagact gttgtcttgt 
               
               
                   
               
               
                 3781 
                 tcttgtggtg tttggattct tgtggtgtgt gcttttagac acagggtaga attagagaca 
               
               
                   
               
               
                 3841 
                 atattggatg tacaattcct caggagacta cagtagtata ttctattcct taccagtaat 
               
               
                   
               
               
                 3901 
                 aaggttcttc ctaataataa ttaagagatt gaaactccaa acaagtattc attatgaaca 
               
               
                   
               
               
                 3961 
                 gatacacatc aaaatcataa taatattttc aaaacaagga ataatttctc taatggttta 
               
               
                   
               
               
                 4021 
                 ttatagaata ccaatgtata gcttagaaat aaaactttga atatttcaag aatatagata 
               
               
                   
               
               
                 4081 
                 agtctaattt ttaaatgctg tatatatggc tttcactcaa tcatctctca gatgttgtta 
               
               
                   
               
               
                 4141 
                 ttaactcgct ctgtgttgtt gcaaaacttt ttggtgcaga ttcgtttcca aaactattgc 
               
               
                   
               
               
                 4201 
                 tactttgtgt gctttaaaca aaataccttg ggttgatgaa acatcaaccc agtgctagga 
               
               
                   
               
               
                 4261 
                 atactgtgta tctatcatta gctatatggg actatattgt agattgtggt ttctcagtag 
               
               
                   
               
               
                 4321 
                 agaagtgact gtagtgtgat tctagataaa tcatcattag caattcattc agatggtcaa 
               
               
                   
               
               
                 4381 
                 taacttgaaa tttatagctg tgataggagt tcagaaattg gcacatccct ttaaaaataa 
               
               
                   
               
               
                 4441 
                 caacagaaaa tacaactcct gggaaaaaag gtgctgattc tataagatta tttatatatg 
               
               
                   
               
               
                 4501 
                 taagtgttta aaaagattat tttccagaaa gtttgtgcag ggtttaagtt gctactattc 
               
               
                   
               
               
                 4561 
                 aactacacta tatataaata aaatatatac aatatataca ttgttttcac tgtatcacat 
               
               
                   
               
               
                 4621 
                 taaagtactt gggcttcaga agtaagagcc aaccaactga aaacctgaga tggagatatg 
               
               
                   
               
               
                 4681 
                 ttcaaagaat gagatacaat tttttagttt tcagtttaag taactctcag cattacaaaa 
               
               
                   
               
               
                 4741 
                 gagtaagtat ctcacaaata ggaaataaaa ctaaaacgtg gatttaaaaa gaactgcacg 
               
               
                   
               
               
                 4801 
                 ggctttaggg taaatgctca tcttaaacct cactagaggg aagtcttctc aagtttcaag 
               
               
                   
               
               
                 4861 
                 caagaccatt tacttaatgt gaagttttgg aaagttataa aggtgtatgt tttagccata 
               
               
                   
               
               
                 4921 
                 tgattttaat tttaattttg cttcttttag gttcgttctt atttaaagca atatgattgt 
               
               
                   
               
               
                 4981 
                 gtgactcctt gtagttacac ttgtgtttca atcagatcag attgttgtat ttattccact 
               
               
                   
               
               
                 5041 
                 attttgcatt taaatgataa cataaaagat ataaaaaatt taaaactgct atttttctta 
               
               
                   
               
               
                 5101 
                 tagaagagaa aatgggtgtt ggtgattgta ttttaattat ttaagcgtct ctgtttacct 
               
               
                   
               
               
                 5161 
                 gcctaggaaa acattttatg gcagtcttat gtgcaaagat cgtaaaagga caaaaaattt 
               
               
                   
               
               
                 5221 
                 aaactgctta taataatcca ggagttgcat tatagccagt agtaaaaata ataataataa 
               
               
                   
               
               
                 5281 
                 taataaaacc atgtctatag ctgtagatgg gcttcacatc tgtaaagcaa tcaattgtat 
               
               
                   
               
               
                 5341 
                 atttttgtga tgtgtaccat actgtgtgct ccagcaaatg tccatttgtg taaatgtatt 
               
               
                   
               
               
                 5401 
                 tattttatat tgtatatatt gttaaatgca aaaaggagat atgattctgt aactccaatc 
               
               
                   
               
               
                 5461 
                 agttcagatg tgtaactcaa attattatgc ctttcaggat gatggtagag caatattaaa 
               
               
                   
               
               
                 5521 
                 caagcttcca cttttgactg ctaaaaaaaa aaaaaaaaa 
               
            
           
         
       
     
     As used herein, “endocrine” refers to secretion of an agent (e.g., a hormone) into a bloodstream. “Exocrine” refers to secretion of an agent into an epithelial surface by way of a duct. 
     By “fragment” is meant a portion of a polypeptide or nucleic acid molecule. This portion contains at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% of the entire length of the reference nucleic acid molecule or polypeptide. A fragment may contain 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100, 200, 300, 400, 500, 600, 700, 800, 900, or 1000 nucleotides or amino acids. 
     By “FOXA2 polypeptide” is meant a protein or fragment thereof having at least 85% amino acid sequence identity to the sequence provided at NCBI Accession No. NP_068556.2 and having transcription factor activity. The amino acid sequence provided at NCBI Accession No. NP_068556.2 is shown below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 11) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 mhsassmlga vkmeghepsd wssyyaepeg yssvsnmnag lgmngmntym smsaaamgsg 
                   
               
               
                   
               
               
                 61 
                 sgnmsagsmn mssyvgagms pslagmspga gamagmggsa gaagvagmgp hlspslsplg 
               
               
                   
               
               
                 121 
                 gqaagamggl apyanmnsms pmygqaglsr ardpktyrrs ythakppysy islitmaigq 
               
               
                   
               
               
                 181 
                 spnkmltlse iyqwimdlfp fyrqnqqrwq nsirhslsfn dcflkvprsp dkpgkgsfwt 
               
               
                   
               
               
                 241 
                 lhpdsgnmfe ngcylrrqkr fkcekqlalk eaagaagsgk kaaagagasq aqlgeaagpa 
               
               
                   
               
               
                 301 
                 setpagtesp hssaspcqeh krgglgelkg tpaaalsppe papspgqqqq aaahllgpph 
               
               
                   
               
               
                 361 
                 hpglppeahl kpehhyafnh pfsinnlmss eqqhhhshhh hqphkmdlka yeqvmhypgy 
               
               
                   
               
               
                 421 
                 gspmpgslam gpvtnktgld asplaadtsy yqgvysrpim nss 
               
            
           
         
       
     
     By “FOXA2 polynucleotide” is meant a polynucleotide encoding a FOXA2 polypeptide or fragment thereof. An exemplary FOXA2 polynucleotide sequence is provided at NCBI Ref: NM_021784.4. The sequence provided at NCBI Ref: NM_021784.4 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 12) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 cccgcccact tccaactacc gcctccggcc tgcccaggga gagagaggga gtggagccca 
                   
               
               
                   
               
               
                 61 
                 gggagaggga gcgcgagaga gggagggagg aggggacggt gctttggctg actttttttt 
               
               
                   
               
               
                 121 
                 aaaagagggt gggggtgggg ggtgattgct ggtcgtttgt tgtggctgtt aaattttaaa 
               
               
                   
               
               
                 181 
                 ctgccatgca ctcggcttcc agtatgctgg gagcggtgaa gatggaaggg cacgagccgt 
               
               
                   
               
               
                 241 
                 ccgactggag cagctactat gcagagcccg agggctactc ctccgtgagc aacatgaacg 
               
               
                   
               
               
                 301 
                 ccggcctggg gatgaacggc atgaacacgt acatgagcat gtcggcggcc gccatgggca 
               
               
                   
               
               
                 361 
                 gcggctcggg caacatgagc gcgggctcca tgaacatgtc gtcgtacgtg ggcgctggca 
               
               
                   
               
               
                 421 
                 tgagcccgtc cctggcgggg atgtcccccg gcgcgggcgc catggcgggc atgggcggct 
               
               
                   
               
               
                 481 
                 cggccggggc ggccggcgtg gcgggcatgg ggccgcactt gagtcccagc ctgagcccgc 
               
               
                   
               
               
                 541 
                 tcggggggca ggcggccggg gccatgggcg gcctggcccc ctacgccaac atgaactcca 
               
               
                   
               
               
                 601 
                 tgagccccat gtacgggcag gcgggcctga gccgcgcccg cgaccccaag acctacaggc 
               
               
                   
               
               
                 661 
                 gcagctacac gcacgcaaag ccgccctact cgtacatctc gctcatcacc atggccatcc 
               
               
                   
               
               
                 721 
                 agcagagccc caacaagatg ctgacgctga gcgagatcta ccagtggatc atggacctct 
               
               
                   
               
               
                 781 
                 tccccttcta ccggcagaac cagcagcgct ggcagaactc catccgccac tcgctctcct 
               
               
                   
               
               
                 841 
                 tcaacgactg tttcctgaag gtgccccgct cgcccgacaa gcccggcaag ggctccttct 
               
               
                   
               
               
                 901 
                 ggaccctgca ccctgactcg ggcaacatgt tcgagaacgg ctgctacctg cgccgccaga 
               
               
                   
               
               
                 961 
                 agcgcttcaa gtgcgagaag cagctggcgc tgaaggaggc cgcaggcgcc gccggcagcg 
               
               
                   
               
               
                 1021 
                 gcaagaaggc ggccgccgga gcccaggcct cacaggctca actcggggag gccgccgggc 
               
               
                   
               
               
                 1081 
                 cggcctccga gactccggcg ggcaccgagt cgcctcactc gagcgcctcc ccgtgccagg 
               
               
                   
               
               
                 1141 
                 agcacaagcg agggggcctg ggagagctga aggggacgcc ggctgcggcg ctgagccccc 
               
               
                   
               
               
                 1201 
                 cagagccggc gccctctccc gggcagcagc agcaggccgc ggcccacctg ctgggcccgc 
               
               
                   
               
               
                 1261 
                 cccaccaccc gggcctgccg cctgaggccc acctgaagcc ggaacaccac tacgccttca 
               
               
                   
               
               
                 1321 
                 accacccgtt ctccatcaac aacctcatgt cctcggagca gcagcaccac cacagccacc 
               
               
                   
               
               
                 1381 
                 accaccacca accccacaaa atggacctca aggcctacga acaggtgatg cactaccccg 
               
               
                   
               
               
                 1441 
                 gctacggttc ccccatgcct ggcagcttgg ccatgggccc ggtcacgaac aaaacgggcc 
               
               
                   
               
               
                 1501 
                 tggacgcctc gcccctggcc gcagatacct cctactacca gggggtgtac tcccggccca 
               
               
                   
               
               
                 1561 
                 ttatgaactc ctcttaagaa gacgacggct tcaggcccgg ctaactctgg caccccggat 
               
               
                   
               
               
                 1621 
                 cgaggacaag tgagagagca agtgggggtc gagactttgg ggagacggtg ttgcagagac 
               
               
                   
               
               
                 1681 
                 gcaagggaga agaaatccat aacaccccca ccccaacacc cccaagacag cagtcttctt 
               
               
                   
               
               
                 1741 
                 cacccgctgc agccgttccg tcccaaacag agggccacac agatacccca cgttctatat 
               
               
                   
               
               
                 1801 
                 aaggaggaaa acgggaaaga atataaagtt aaaaaaaagc ctccggtttc cactactgtg 
               
               
                   
               
               
                 1861 
                 tagactcctg cttcttcaag cacctgcaga ttctgatttt tttgttgttg ttgttctcct 
               
               
                   
               
               
                 1921 
                 ccattgctgt tgttgcaggg aagtcttact taaaaaaaaa aaaaaatttt gtgagtgact 
               
               
                   
               
               
                 1981 
                 cggtgtaaaa ccatgtagtt ttaacagaac cagagggttg tactattgtt taaaaacagg 
               
               
                   
               
               
                 2041 
                 aaaaaaaata atgtaagggt ctgttgtaaa tgaccaagaa aaagaaaaaa aaagcattcc 
               
               
                   
               
               
                 2101 
                 caatcttgac acggtgaaat ccaggtctcg ggtccgatta atttatggtt tctgcgtgct 
               
               
                   
               
               
                 2161 
                 ttatttatgg cttataaatg tgtattctgg ctgcaagggc cagagttcca caaatctata 
               
               
                   
               
               
                 2221 
                 ttaaagtgtt atacccggtt ttatcccttg aatcttttct tccagatttt tcttttcttt 
               
               
                   
               
               
                 2281 
                 acttggctta caaaatatac aggcttggaa attatttcaa gaaggaggga gggataccct 
               
               
                   
               
               
                 2341 
                 gtctggttgc aggttgtatt ttattttggc ccagggagtg ttgctgtttt cccaacattt 
               
               
                   
               
               
                 2401 
                 tattaataaa attttcagac ataaaaaa 
               
            
           
         
       
     
     By “GATA6 polypeptide” is meant a protein or fragment thereof having at least 85% amino acid sequence identity to the sequence provided at NCBI Accession No. NP_005248.2 and having transcription factor activity. The amino acid sequence provided at NCBI Accession No. NP_005248.2 is shown below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 13) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 maltdggwcl pkrfgaagad asdsrafpar epstppspis ssssscsrgg ergpggasnc 
                   
               
               
                   
               
               
                 61 
                 gtpqldteaa agpparslll ssyashpfga phgpsapgva gpggnlsswe dlllftdldq 
               
               
                   
               
               
                 121 
                 aataskllws srgaklspfa peqpeemyqt laalssqgpa aydgapggfv hsaaaaaaaa 
               
               
                   
               
               
                 181 
                 aaasspvyvp ttrvgsmlpg lpyhlqgsgs gpanhaggag ahpgwpqasa dsppygsggg 
               
               
                   
               
               
                 241 
                 aagggaagpg gagsaaahvs arfpyspspp mangaarepg gyaaagsgga ggvsgggssl 
               
               
                   
               
               
                 301 
                 aamggrepqy sslsaarpln gtyhhhhhhh hhhpspyspy vgapltpawp agpfetpvlh 
               
               
                   
               
               
                 361 
                 slqsragapl pvprgpsadl ledlsesrec vncgsiqtpl wrrdgtghyl cnacglyskm 
               
               
                   
               
               
                 421 
                 nglsrplikp qkrvpssrrl glscanchtt tttlwrrnae gepvcnacgl ymklhgvprp 
               
               
                   
               
               
                 481 
                 lamkkegiqt rkrkpknink sktcsgnsnn sipmtptsts snsddcsknt spttqptasg 
               
               
                   
               
               
                 541 
                 agapvmtgag estnpensel kysgqdglyi gvslaspaev tssvrpdswc alala 
               
            
           
         
       
     
     By “GATA6 polynucleotide” is meant a polynucleotide encoding a GATA6 polypeptide or fragment thereof. An exemplary KCNK3 polynucleotide sequence is provided at NCBI Ref: NM_005257.5. The sequence provided at NCBI Ref: NM_005257.5 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 14) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 agttccgacc cacagcctgg cacccttcgg cgagcgctgt ttgtttaggg ctcggtgagt 
                   
               
               
                   
               
               
                 61 
                 ccaatcagga gcccaggctg cagttttccg gcagagcagt aagaggcgcc tcctctctcc 
               
               
                   
               
               
                 121 
                 tttttattca ccagcagcgc ggcgcagacc ccggactcgc gctcgcccgc tggcgccctc 
               
               
                   
               
               
                 181 
                 ggcttctctc cgcgcctggg agcaccctcc gccgcggccg ttctccatgc gcagcgcccg 
               
               
                   
               
               
                 241 
                 cccgaggagc tagacgtcag cttggagcgg cgccggaccg tggatggcct tgactgacgg 
               
               
                   
               
               
                 301 
                 cggctggtgc ttgccgaagc gcttcggggc cgcgggtgcg gacgccagcg actccagagc 
               
               
                   
               
               
                 361 
                 ctttccagcg cgggagccct ccacgccgcc ttcccccatc tcttcctcgt cctcctcctg 
               
               
                   
               
               
                 421 
                 ctcccggggc ggagagcggg gccccggcgg cgccagcaac tgcgggacgc ctcagctcga 
               
               
                   
               
               
                 481 
                 cacggaggcg gcggccggac ccccggcccg ctcgctgctg ctcagttcct acgcttcgca 
               
               
                   
               
               
                 541 
                 tcccttcggg gctccccacg gaccttcggc gcctggggtc gcgggccccg ggggcaacct 
               
               
                   
               
               
                 601 
                 gtcgagctgg gaggacttgc tgctgttcac tgacctcgac caagccgcga ccgccagcaa 
               
               
                   
               
               
                 661 
                 gctgctgtgg tccagccgcg gcgccaagct gagccccttc gcacccgagc agccggagga 
               
               
                   
               
               
                 721 
                 gatgtaccag accctcgccg ctctctccag ccagggtccg gccgcctacg acggcgcgcc 
               
               
                   
               
               
                 781 
                 cggcggcttc gtgcactctg cggccgcggc ggcagcagcc gcggcggcgg ccagctcccc 
               
               
                   
               
               
                 841 
                 ggtctacgtg cccaccaccc gcgtgggttc catgctgccc ggcctaccgt accacctgca 
               
               
                   
               
               
                 901 
                 ggggtcgggc agtgggccag ccaaccacgc gggcggcgcg ggcgcgcacc ccggctggcc 
               
               
                   
               
               
                 961 
                 tcaggcctcg gccgacagcc ctccatacgg cagcggaggc ggcgcggctg gcggcggggc 
               
               
                   
               
               
                 1021 
                 cgcggggcct ggcggcgctg gctcagccgc ggcgcacgtc tcggcgcgct tcccctactc 
               
               
                   
               
               
                 1081 
                 tcccagcccg cccatggcca acggcgccgc gcgggagccg ggaggctacg cggcggcggg 
               
               
                   
               
               
                 1141 
                 cagtgggggc gcgggaggcg tgagcggcgg cggcagtagc ctggcggcca tgggcggccg 
               
               
                   
               
               
                 1201 
                 cgagccccag tacagctcgc tgtcggccgc gcggccgctg aacgggacgt accaccacca 
               
               
                   
               
               
                 1261 
                 ccaccaccac caccaccacc atccgagccc ctactcgccc tacgtggggg cgccactgac 
               
               
                   
               
               
                 1321 
                 gcctgcctgg cccgccggac ccttcgagac cccggtgctg cacagcctgc agagccgcgc 
               
               
                   
               
               
                 1381 
                 cggagccccg ctcccggtgc cccggggtcc cagtgcagac ctgctggagg acctgtccga 
               
               
                   
               
               
                 1441 
                 gagccgcgag tgcgtgaact gcggctccat ccagacgccg ctgtggcggc gggacggcac 
               
               
                   
               
               
                 1501 
                 cggccactac ctgtgcaacg cctgcgggct ctacagcaag atgaacggcc tcagccggcc 
               
               
                   
               
               
                 1561 
                 cctcatcaag ccgcagaagc gcgtgccttc atcacggcgg cttggattgt cctgtgccaa 
               
               
                   
               
               
                 1621 
                 ctgtcacacc acaactacca ccttatggcg cagaaacgcc gagggtgaac ccgtgtgcaa 
               
               
                   
               
               
                 1681 
                 tgcttgtgga ctctacatga aactccatgg ggtgcccaga ccacttgcta tgaaaaaaga 
               
               
                   
               
               
                 1741 
                 gggaattcaa accaggaaac gaaaacctaa gaacataaat aaatcaaaga cttgctctgg 
               
               
                   
               
               
                 1801 
                 taatagcaat aattccattc ccatgactcc aacttccacc tcttctaact cagatgattg 
               
               
                   
               
               
                 1861 
                 cagcaaaaat acttccccca caacacaacc tacagcctca ggggcgggtg ccccggtgat 
               
               
                   
               
               
                 1921 
                 gactggtgcg ggagagagca ccaatcccga gaacagcgag ctcaagtatt cgggtcaaga 
               
               
                   
               
               
                 1981 
                 tgggctctac ataggcgtca gtctcgcctc gccggccgaa gtcacgtcct ccgtgcgacc 
               
               
                   
               
               
                 2041 
                 ggattcctgg tgcgccctgg ccctggcctg agcccacgcc gccaggaggc agggagggct 
               
               
                   
               
               
                 2101 
                 ccgccgcggg cctcactcca ctcgtgtctg cttttgtgca gcggtccaga cagtggcgac 
               
               
                   
               
               
                 2161 
                 tgcgctgaca gaacgtgatt ctcgtgcctt tattttgaaa gagatgtttt tcccaagagg 
               
               
                   
               
               
                 2221 
                 cttgctgaaa gagtgagaga agatggaagg gaagggccag tgcaactggg cgcttgggcc 
               
               
                   
               
               
                 2281 
                 actccagcca gcccgcctcc ggggcggacc ctgctccact tccagaagcc aggactagga 
               
               
                   
               
               
                 2341 
                 cctgggcctt gcctgctatg gaatattgag agagattttt taaaaaagat tttgcatttt 
               
               
                   
               
               
                 2401 
                 gtccaaaatc atgtgcttct tctgatcaat tttggttgtt ccagaatttc ttcatacctt 
               
               
                   
               
               
                 2461 
                 ttccacatcc agatttcatg tgcgttcatg gagaagatca cttgaggcca tttggtacac 
               
               
                   
               
               
                 2521 
                 atctctggag gctgagtcgg ttcatgaggt ctcttatcaa aaatattact cagtttgcaa 
               
               
                   
               
               
                 2581 
                 gactgcattg taactttaac atacactgtg actgacgttt ctcaaagttc atattgtgtg 
               
               
                   
               
               
                 2641 
                 gctgatctga agtcagtcgg aatttgtaaa cagggtagca aacaagatat ttttcttcca 
               
               
                   
               
               
                 2701 
                 tgtatacaat aattttttta aaaagtgcaa tttgcgttgc agcaatcagt gttaaatcat 
               
               
                   
               
               
                 2761 
                 ttgcataaga tttaacagca ttttttataa tgaatgtaaa cattttaact taatggtact 
               
               
                   
               
               
                 2821 
                 taaaataatt taaaagaaaa atgttaactt agacattctt atgcttcttt tacaactaca 
               
               
                   
               
               
                 2881 
                 tcccatttta tatttccaat tgttaaagaa aaatatttca agaacaaatc ttctctcagg 
               
               
                   
               
               
                 2941 
                 aaaattgcct ttctctattt gttaagaatt tttatacaag aacaccaata tacccccttt 
               
               
                   
               
               
                 3001 
                 attttactgt ggaatatgtg ctggaaaaat tgcaacaaca ctttactacc taacggatag 
               
               
                   
               
               
                 3061 
                 catttgtaaa tactctaggt atctgtaaac actctgatga agtctgtata gtgtgactaa 
               
               
                   
               
               
                 3121 
                 cccacaggca ggttggttta cattaatttt tttttttgaa tgggatgtcc tatggaaacc 
               
               
                   
               
               
                 3181 
                 tatttcacca gagttttaaa aataaaaagg gtattgtttt gtcttctgta cagtgagttc 
               
               
                   
               
               
                 3241 
                 cttccctttt caaagctttc tttttatgct gtatgtgact atagatattc atataaaaca 
               
               
                   
               
               
                 3301 
                 agtgcacgtg aagtttgcaa aatgctttaa ggccttcctt tcaaagcata gtccttttgg 
               
               
                   
               
               
                 3361 
                 agccgttttg taccttttat accttggctt atttgaagtt gacacatggg gttagttact 
               
               
                   
               
               
                 3421 
                 actctccatg tgcattgggg acagttttta taagtgggaa ggactcagta ttattatatt 
               
               
                   
               
               
                 3481 
                 tgagatgata agcattttgt ttgggaacaa tgcttaaaaa tattccagaa agttcagatt 
               
               
                   
               
               
                 3541 
                 ttttttcttt gtgaatgaaa tatattctgg cccacgaaca gggcgatttc ctttcagttt 
               
               
                   
               
               
                 3601 
                 tttccttttg caacgtgcct tgaagtctca aagctcacct gaggttgcag acgttacccc 
               
               
                   
               
               
                 3661 
                 caacagaaga taggtagaaa tgattccagt ggcctctttg tattttcttc attgttgagt 
               
               
                   
               
               
                 3721 
                 agatttcagg aaatcaggag gtgtttcaca atacagaatg atggccttta actgtgaaaa 
               
               
                   
               
               
                 3781 
                 aaaaa 
               
            
           
         
       
     
     By “gellan gum” is meant a polysaccharide having a straight chain with a repeating unit that has any one of the following molecular structures: 
     
       
         
         
             
             
         
       
     
     In the foregoing structures, “Ac” refers to an acetate group and “Gly” refers to a glycerate group and “M+” is a monovalent cation. In some embodiments, the gellan gum is KELCOGEL® gellan gum. 
     “Hybridization” means hydrogen bonding, which may be Watson-Crick, Hoogsteen or reversed Hoogsteen hydrogen bonding, between complementary nucleobases. For example, adenine and thymine are complementary nucleobases that pair through the formation of hydrogen bonds. 
     By “immunosuppressive agent” or “immunosuppressant” is meant an agent that inhibits or prevents an immune reaction, such as rejection, of a transplanted organ or organoid in a subject. Examples of immunosuppressants include, but are not limited to, basilizimab, antithymocyte globulin, alemtuzumab, prednisone, azathioprine, mycophenolate, cyclosporine, sirolimus, methotrexate, interferon, and tacrolimus. 
     By “induced pluripotent stem cell” or “iPSC” is meant a differentiated somatic cell that acquires pluripotency by the exogenous expression of one or more transcription factors in the cell. An “iPSC-derived cell” is a cell derived from an induced pluripotent stem cell. An “iPSC-derived beta-like cell,” “iPSC-derived alpha-like cell,” “iPSC-derived delta-like cell,” or “iPSC-derived duct-like cell” is a cell derived from an induced pluripotent stem cell and has characteristics of a beta cell, alpha cell, delta cell, or duct cell, respectively. 
     The terms “isolated,” “purified,” or “biologically pure” refer to material that is free to varying degrees from components which normally accompany it as found in its native state. “Isolate” denotes a degree of separation from original source or surroundings. “Purify” denotes a degree of separation that is higher than isolation. A “purified” or “biologically pure” protein is sufficiently free of other materials such that any impurities do not materially affect the biological properties of the protein or cause other adverse consequences. That is, a nucleic acid or peptide of this invention is purified if it is substantially free of cellular material, viral material, or culture medium when produced by recombinant DNA techniques, or chemical precursors or other chemicals when chemically synthesized. Purity and homogeneity are typically determined using analytical chemistry techniques, for example, polyacrylamide gel electrophoresis or high performance liquid chromatography. The term “purified” can denote that a nucleic acid or protein gives rise to essentially one band in an electrophoretic gel. For a protein that can be subjected to modifications, for example, phosphorylation or glycosylation, different modifications may give rise to different isolated proteins, which can be separately purified. 
     By “isolated polynucleotide” is meant a nucleic acid (e.g., a DNA) that is free of the genes which, in the naturally-occurring genome of the organism from which the nucleic acid molecule of the invention is derived, flank the gene. The term therefore includes, for example, a recombinant DNA that is incorporated into a vector; into an autonomously replicating plasmid or virus; or into the genomic DNA of a prokaryote or eukaryote; or that exists as a separate molecule (for example, a cDNA or a genomic or cDNA fragment produced by PCR or restriction endonuclease digestion) independent of other sequences. In addition, the term includes an RNA molecule that is transcribed from a DNA molecule, as well as a recombinant DNA that is part of a hybrid gene encoding additional polypeptide sequence. 
     By an “isolated polypeptide” is meant a polypeptide of the invention that has been separated from components that naturally accompany it. Typically, the polypeptide is isolated when it is at least 60%, by weight, free from the proteins and naturally-occurring organic molecules with which it is naturally associated. The preparation can be at least 75%, at least 90%, and at least 99%, by weight, a polypeptide of the invention. An isolated polypeptide of the invention may be obtained, for example, by extraction from a natural source, by expression of a recombinant nucleic acid encoding such a polypeptide; or by chemically synthesizing the protein. Purity can be measured by any appropriate method, for example, column chromatography, polyacrylamide gel electrophoresis, or by HPLC analysis. 
     By “KCNK3 polypeptide” is meant a protein or fragment thereof having at least 85% amino acid sequence identity to the sequence provided at NCBI Accession No. NP_002237.1 and having potassium channel activity. The amino acid sequence provided at NCBI Accession No. NP_002237.1 is shown below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 15) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 mkrqnvrtla livctftyll vgaavfdale sepelierqr lelrqqelra rynlsqggye 
                   
               
               
                   
               
               
                 61 
                 elervvlrlk phkagvqwrf agsfyfaitv ittigyghaa pstdggkvfc mfyallgipl 
               
               
                   
               
               
                 121 
                 tlvmfqslge rintlvryll hrakkglgmr radvsmanmv ligffscist lcigaaafsh 
               
               
                   
               
               
                 181 
                 yehwtffqay yycfitltti gfgdyvalqk dgalgtqpqy vafsfvyilt gltvigafln 
               
               
                   
               
               
                 241 
                 lvvlrfmtmn aedekrdaeh ralltrngqa gggggggsah ttdtasstaa aggggfrnvy 
               
               
                   
               
               
                 301 
                 aevlhfqsmc sclwyksrek lqysipmiip rdlstsdtcv eqshsspggg grysdtpsrr 
               
               
                   
               
               
                 361 
                 clcsgaprsa issystglhs lstfrglmkr rssv 
               
            
           
         
       
     
     By “KCNK3 polynucleotide” is meant a polynucleotide encoding a KCNK3 polypeptide or fragment thereof. An exemplary KCNK3 polynucleotide sequence is provided at NCBI Ref: NM_002246.2. The sequence provided at NCBI Ref: NM_002246.2 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 16) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 ggcggcggcg gcggcggcgg ccccgggcgc tgagcgggtg cccggcgcgg agagcggcga 
                   
               
               
                   
               
               
                 61 
                 gcgcagccat gccccaggcc gcctccgggg cagcagcagc ggcggccggg gccgaggcgc 
               
               
                   
               
               
                 121 
                 gggccggggg cgccgggggg ccggcggcgg cccgggcggg acgatgaagc ggcagaacgt 
               
               
                   
               
               
                 181 
                 gcgcacgctg gcgctcatcg tgtgcacctt cacctacctg ctggtgggcg ccgcggtctt 
               
               
                   
               
               
                 241 
                 cgacgcgctg gagtcggagc ccgagctgat cgagcggcag cggctggagc tgcggcagca 
               
               
                   
               
               
                 301 
                 ggagctgcgg gcgcgctaca acctcagcca gggcggctac gaggagctgg agcgcgtcgt 
               
               
                   
               
               
                 361 
                 gctgcgcctc aagccgcaca aggccggcgt gcagtggcgc ttcgccggct ccttctactt 
               
               
                   
               
               
                 421 
                 cgccatcacc gtcatcacca ccatcggcta cgggcacgcg gcacccagca cggatggcgg 
               
               
                   
               
               
                 481 
                 caaggtgttc tgcatgttct acgcgctgct gggcatcccg ctcacgctcg tcatgttcca 
               
               
                   
               
               
                 541 
                 gagcctgggc gagcgcatca acaccttggt gaggtacctg ctgcaccgcg ccaagaaggg 
               
               
                   
               
               
                 601 
                 gctgggcatg cggcgcgccg acgtgtccat ggccaacatg gtgctcatcg gcttcttctc 
               
               
                   
               
               
                 661 
                 gtgcatcagc acgctgtgca tcggcgccgc cgccttctcc cactacgagc actggacctt 
               
               
                   
               
               
                 721 
                 cttccaggcc tactactact gcttcatcac cctcaccacc atcggcttcg gcgactacgt 
               
               
                   
               
               
                 781 
                 ggcgctgcag aaggaccagg ccctgcagac gcagccgcag tacgtggcct tcagcttcgt 
               
               
                   
               
               
                 841 
                 ctacatcctt acgggcctca cggtcatcgg cgccttcctc aacctcgtgg tgctgcgctt 
               
               
                   
               
               
                 901 
                 catgaccatg aacgccgagg acgagaagcg cgacgccgag caccgcgcgc tgctcacgcg 
               
               
                   
               
               
                 961 
                 caacgggcag gcgggcggcg gcggaggggg tggcagcgcg cacactacgg acaccgcctc 
               
               
                   
               
               
                 1021 
                 atccacggcg gcagcgggcg gcggcggctt ccgcaacgtc tacgcggagg tgctgcactt 
               
               
                   
               
               
                 1081 
                 ccagtccatg tgctcgtgcc tgtggtacaa gagccgcgag aagctgcagt actccatccc 
               
               
                   
               
               
                 1141 
                 catgatcatc ccgcgggacc tctccacgtc cgacacgtgc gtggagcaga gccactcgtc 
               
               
                   
               
               
                 1201 
                 gccgggaggg ggcggccgct acagcgacac gccctcgcga cgctgcctgt gcagcggggc 
               
               
                   
               
               
                 1261 
                 gccacgctcc gccatcagct cggtgtccac gggtctgcac agcctgtcca ccttccgcgg 
               
               
                   
               
               
                 1321 
                 cctcatgaag cgcaggagct ccgtgtgact gccccgaggg gcctggagca cctgggggcg 
               
               
                   
               
               
                 1381 
                 cgggcggggg acccctgctg ggaggccagg agactgcccc tgctgccttc tgcccagtgg 
               
               
                   
               
               
                 1441 
                 gaccccgcac aacatccctc accactctcc cccagcaccc ccatctccga ctgtgcctgc 
               
               
                   
               
               
                 1501 
                 ttgcaccagc cggcaggagg ccgggctctg aggacccctg gggcccccat cggagccctg 
               
               
                   
               
               
                 1561 
                 caaattccga gaaatgtgaa acttggtggg gtcagggagg aaaggcagaa gctgggagcc 
               
               
                   
               
               
                 1621 
                 tcccttccct ttgaaaatct aagaagctcc cagtcctcag agaccctgct ggtacccaga 
               
               
                   
               
               
                 1681 
                 cccccacctt cggaggggac ttcatgttcc gtgtacgttt gcatctctat ttatacctct 
               
               
                   
               
               
                 1741 
                 gtcctgctag gtctcccacc ttcccttggt tccaaaagcc agggtgtcta tgtccaagtc 
               
               
                   
               
               
                 1801 
                 acccctactc agccccactc cccttcctca tccccagctg tgtctcccaa cctcccttcg 
               
               
                   
               
               
                 1861 
                 tgttgttttg catggctttg cagttatgga gaaagtggaa acccagcagt ccctaaagct 
               
               
                   
               
               
                 1921 
                 ggtccccaga aagcaggaca gaaagaagga gggacaggca ggcagcagga ggggcgagct 
               
               
                   
               
               
                 1981 
                 gggaggcagg aggcagcggc ctgtcagtct gcagaatggt cgcactggag gttcaagcta 
               
               
                   
               
               
                 2041 
                 actggcctcc agccacattc tcatagcagg taggacttca gccttccaga cactgccctt 
               
               
                   
               
               
                 2101 
                 agaatctgga acagaagact tcagactcac cataattgct gataattacc cactcttaaa 
               
               
                   
               
               
                 2161 
                 tttgtcgagt gatttttagc ctctgaaaac tctatgctgg ccactgattc ctttgagtct 
               
               
                   
               
               
                 2221 
                 cacaaaaccc tacttaggtc atcagggcag gagttctcac tcccatttta cagatgagaa 
               
               
                   
               
               
                 2281 
                 tactgaggcc tggacaggtg aagtgaccag agagcaaaag gcaaaggggt gggggctggg 
               
               
                   
               
               
                 2341 
                 tgcagtggct cacacctgta ttcccaacac ttttggaggc tgaggttgga ggattgcttg 
               
               
                   
               
               
                 2401 
                 agcccaggaa tttgagacca gcctaggtga catagtgaga ccccatctct acaaaaaata 
               
               
                   
               
               
                 2461 
                 aaaaattaac caggtgtggt ggcacgtgcc tgggagtccc agcgacttgg gaggctgagg 
               
               
                   
               
               
                 2521 
                 tgggaggatt gtttgagcct gggaggtcga ggctgtagtg agccctgatt gcaccactgt 
               
               
                   
               
               
                 2581 
                 actccagcct gggtgacagg gcaagaccct gtctcaaaaa aaaaaaaaaa aatggcaaag 
               
               
                   
               
               
                 2641 
                 ggagacaaga gcccagcctg cttgttgcta gccaaagtgt tctttccttc cagcttggcc 
               
               
                   
               
               
                 2701 
                 tgctcttaaa agcaaagctc ctgcagtgta catcctggca ttgtgtggct acctgggttt 
               
               
                   
               
               
                 2761 
                 taaaccagaa tcagaagtcc cggatcagag ggcactgctg aggttcagcc tcttctcttc 
               
               
                   
               
               
                 2821 
                 ttggccagga ggcagcagct ctgaatgggc ccctgaggct gcacaggggc ctttgtcact 
               
               
                   
               
               
                 2881 
                 ggggcgcatg cttacaaaca gtgcagttct tgggaccgag gtaagcaggg ctgggtctca 
               
               
                   
               
               
                 2941 
                 tggcagaaag gccaggatct ggggctctag gaatttggga attgggcaga gtggccaaga 
               
               
                   
               
               
                 3001 
                 aagctggcag gcatatccta tgggacatca cacctggcac cattgtcatt gttggtgcct 
               
               
                   
               
               
                 3061 
                 gtgtcccaag tagctagtga taagctgagg ctgcagcaag aaacaccctt cccaggtggg 
               
               
                   
               
               
                 3121 
                 ggagtttgga ccagaggtgc cctctgccca ccacacctgc aacccagaag cccagatgga 
               
               
                   
               
               
                 3181 
                 acgcagctga cgaaggtgat gcttgaggct cacttttggg gccccacagc tggagccggt 
               
               
                   
               
               
                 3241 
                 ataatgactg ggacaacatc aaggggtgga tgaggggcct ctcctcccgc aacactgcct 
               
               
                   
               
               
                 3301 
                 tcccatgctg ttcccctgcc agctccttaa cactgccgac caaggccagc cctggcattc 
               
               
                   
               
               
                 3361 
                 agggaaattg gagggcagca cccgtagggt ggccagcctc aggccccacc ccagctgtgt 
               
               
                   
               
               
                 3421 
                 cctctagtct ctggggaccc ctggggggaa gaagtctacc ctgcttgtga gtcccgtctc 
               
               
                   
               
               
                 3481 
                 agtgtggagg aactggctgc acgtgggacc tgaaggtgcc ctctgtgttt atgttggggg 
               
               
                   
               
               
                 3541 
                 tgggggggca gtgctggctg cctctgtcct gtgtgtgacc ctgccctcga agggtcctgt 
               
               
                   
               
               
                 3601 
                 cctgtcagtc ccgagggagc cacaaccaaa gctgcggaga gaaggtgggg aagggtgcag 
               
               
                   
               
               
                 3661 
                 aatggccgtg gggcacagcg tggcagactg ttcagtctct gctgggtctt tcctagggac 
               
               
                   
               
               
                 3721 
                 ctggaaggcc agtgttgctt ccccctcact ccctttcact gcaggcagcc tctctgcttc 
               
               
                   
               
               
                 3781 
                 cccaatgcct tatgcctggg cacactgcca cagaatatgc aatatgtgtg ggtgaccatg 
               
               
                   
               
               
                 3841 
                 ccctcacgac cacaccccca ccccgggcag cccccggact ccaaaggtcg tggctgccac 
               
               
                   
               
               
                 3901 
                 agcctccctc agctcttcct gcctatctgt cttcacactg agaatggcgc ccaataaatg 
               
               
                   
               
               
                 3961 
                 ctatccacgg agaccagg 
               
            
           
         
       
     
     By “KCNQ1 polypeptide” is meant a protein or fragment thereof having at least 85% amino acid sequence identity to the sequence provided at NCBI Accession No. NP_000209.2 (isoform 1) or NP_861463.1 (isoform 2) and having potassium channel activity. The amino acid sequence provided at NCBI Accession No. NP_000209.2 is shown below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 17) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 maaassppra erkrwgwgrl pgarrgsagl akkcpfslel aeggpaggal yapiapgapg 
                   
               
               
                   
               
               
                 61 
                 pappaspaap aappvasdlg prppvsldpr vsiystrrpv larthvqgry ynflerptgw 
               
               
                   
               
               
                 121 
                 kcfvyhfavf livlvclifs vlstieqyaa latgtlfwme ivlvvffgte yvvrlwsagc 
               
               
                   
               
               
                 181 
                 rskyvglwgr lrfarkpisi idlivvvasm vvlcvgskgq vfatsairgi rflqilrmlh 
               
               
                   
               
               
                 241 
                 vdrqggtwrl lgsvvfihrq elittlyigf lglifssyfv ylaekdavne sgrvefgsya 
               
               
                   
               
               
                 301 
                 dalwwgvvtv ttigygdkvp qtwvgktias cfsvfaisff alpagilgsg falkvqqkqr 
               
               
                   
               
               
                 361 
                 qkhfnrqipa aasliqtawr cyaaenpdss twkiyirkap rshtllspsp kpkksvvvkk 
               
               
                   
               
               
                 421 
                 kkfkldkdng vtpgekmltv phitcdppee rrldhfsvdg ydssvrkspt llevsmphfm 
               
               
                   
               
               
                 481 
                 rtnsfaedld legetlltpi thisqlrehh ratikvirrm qyfvakkkfq qarkpydvrd 
               
               
                   
               
               
                 541 
                 vieqysqghl nlmvrikelq rrldqsigkp slfisvseks kdrgsntiga rlnrvedkvt 
               
               
                   
               
               
                 601 
                 qldqrlalit dmlhqllslh ggstpgsggp preggahitq pcgsggsvdp elflpsntlp 
               
               
                   
               
               
                 661 
                 tyeqltvprr gpdegs 
               
            
           
         
       
     
     By “KCNQ1 polynucleotide” is meant a polynucleotide encoding a KCNQ1 polypeptide or fragment thereof. An exemplary KCNQ1 polynucleotide sequence is provided at NCBI Ref: NM_000218.2. The sequence provided at NCBI Ref: NM_000218.2 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 18) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 gcggcggggc tggcagcagt ggctgcccgc actgcgcccg ggcgctcgcc ttcgctgcag 
                   
               
               
                   
               
               
                 61 
                 ctcccggtgc cgccgctcgg gccggccccc cggcaggccc tcctcgttat ggccgcggcc 
               
               
                   
               
               
                 121 
                 tcctccccgc ccagggccga gaggaagcgc tggggttggg gccgcctgcc aggcgcccgg 
               
               
                   
               
               
                 181 
                 cggggcagcg cgggcctggc caagaagtgc cccttctcgc tggagctggc ggagggcggc 
               
               
                   
               
               
                 241 
                 ccggcgggcg gcgcgctcta cgcgcccatc gcgcccggcg ccccaggtcc cgcgccccct 
               
               
                   
               
               
                 301 
                 gcgtccccgg ccgcgcccgc cgcgccccca gttgcctccg accttggccc gcggccgccg 
               
               
                   
               
               
                 361 
                 gtgagcctag acccgcgcgt ctccatctac agcacgcgcc gcccggtgtt ggcgcgcacc 
               
               
                   
               
               
                 421 
                 cacgtccagg gccgcgtcta caacttcctc gagcgtccca ccggctggaa atgcttcgtt 
               
               
                   
               
               
                 481 
                 taccacttcg ccgtcttcct catcgtcctg gtctgcctca tcttcagcgt gctgtccacc 
               
               
                   
               
               
                 541 
                 atcgagcagt atgccgccct ggccacgggg actctcttct ggatggagat cgtgctggtg 
               
               
                   
               
               
                 601 
                 gtgttcttcg ggacggagta cgtggtccgc ctctggtccg ccggctgccg cagcaagtac 
               
               
                   
               
               
                 661 
                 gtgggcctct gggggcggct gcgctttgcc cggaagccca tttccatcat cgacctcatc 
               
               
                   
               
               
                 721 
                 gtggtcgtgg cctccatggt ggtcctctgc gtgggctcca aggggcaggt gtttgccacg 
               
               
                   
               
               
                 781 
                 tcggccatca ggggcatccg cttcctgcag atcctgagga tgctacacgt cgaccgccag 
               
               
                   
               
               
                 841 
                 ggaggcacct ggaggctcct gggctccgtg gtcttcatcc accgccagga gctgataacc 
               
               
                   
               
               
                 901 
                 accctgtaca tcggcttcct gggcctcatc ttctcctcgt actttgtgta cctggctgag 
               
               
                   
               
               
                 961 
                 aaggacgcgg tgaacgagtc aggccgcgtg gagttcggca gctacgcaga tgcgctgtgg 
               
               
                   
               
               
                 1021 
                 tggggggtgg tcacagtcac caccatcggc tatggggaca aggtgcccca gacgtgggtc 
               
               
                   
               
               
                 1081 
                 gggaagacca tcgcctcctg cttctctgtc tttgccatct ccttctttgc gctcccagcg 
               
               
                   
               
               
                 1141 
                 gggattcttg gctcggggtt tgccctgaag gtgcagcaga agcagaggca gaagcacttc 
               
               
                   
               
               
                 1201 
                 aaccggcaga tcccggcggc agcctcactc attcagaccg catggaggtg ctatgctgcc 
               
               
                   
               
               
                 1261 
                 gagaaccccg actcctccac ctggaagatc tacatccgga aggccccccg gagccacact 
               
               
                   
               
               
                 1321 
                 ctgctgtcac ccagccccaa acccaagaag tctgtggtgg taaagaaaaa aaagttcaag 
               
               
                   
               
               
                 1381 
                 ctggacaaag acaatggggt gactcctgga gagaagatgc tcacagtccc ccatatcacg 
               
               
                   
               
               
                 1441 
                 tgcgaccccc cagaagagcg gcggctggac cacttctctg tcgacggcta tgacagttct 
               
               
                   
               
               
                 1501 
                 gtaaggaaga gcccaacact gctggaagtg agcatgcccc atttcatgag aaccaacagc 
               
               
                   
               
               
                 1561 
                 ttcgccgagg acctggacct ggaaggggag actctgctga cacccatcac ccacatctca 
               
               
                   
               
               
                 1621 
                 cagctgcggg aacaccatcg ggccaccatt aaggtcattc gacgcatgca gtactttgtg 
               
               
                   
               
               
                 1681 
                 gccaagaaga aattccagca agcgcggaag ccttacgatg tgcgggacgt cattgagcag 
               
               
                   
               
               
                 1741 
                 tactcgcagg gccacctcaa cctcatggtg cgcatcaagg agctgcagag gaggctggac 
               
               
                   
               
               
                 1801 
                 cagtccattg ggaagccctc actgttcatc tccgtctcag aaaagagcaa ggatcgcggc 
               
               
                   
               
               
                 1861 
                 agcaacacga tcggcgcccg cctgaaccga gtagaagaca aggtgacgca gctggaccag 
               
               
                   
               
               
                 1921 
                 aggctggcac tcatcaccga catgcttcac cagctgctct ccttgcacgg tggcagcacc 
               
               
                   
               
               
                 1981 
                 cccggcagcg gcggcccccc cagagagggc ggggcccaca tcacccagcc ctgcggcagt 
               
               
                   
               
               
                 2041 
                 ggcggctccg tcgaccctga gctcttcctg cccagcaaca ccctgcccac ctacgagcag 
               
               
                   
               
               
                 2101 
                 ctgaccgtgc ccaggagggg ccccgatgag gggtcctgag gaggggatgg ggctggggga 
               
               
                   
               
               
                 2161 
                 tgggcctgag tgagagggga ggccaagagt ggccccacct ggccctctct gaaggaggcc 
               
               
                   
               
               
                 2221 
                 acctcctaaa aggcccagag agaagagccc cactctcaga ggccccaata ccccatggac 
               
               
                   
               
               
                 2281 
                 catgctgtct ggcacagcct gcacttgggg gctcagcaag gccacctctt cctggccggt 
               
               
                   
               
               
                 2341 
                 gtgggggccc cgtctcaggt ctgagttgtt accccaagcg ccctggcccc cacatggtga 
               
               
                   
               
               
                 2401 
                 tgttgacatc actggcatgg tggttgggac ccagtggcag ggcacagggc ctggcccatg 
               
               
                   
               
               
                 2461 
                 tatggccagg aagtagcaca ggctgagtgc aggcccaccc tgcttggccc agggggcttc 
               
               
                   
               
               
                 2521 
                 ctgaggggag acagagcaac ccctggaccc cagcctcaaa tccaggaccc tgccaggcac 
               
               
                   
               
               
                 2581 
                 aggcagggca ggaccagccc acgctgacta cagggccgcc ggcaataaaa gcccaggagc 
               
               
                   
               
               
                 2641 
                 ccatttggag ggcctgggcc tggctccctc actctcagga aatgctgacc catgggcagg 
               
               
                   
               
               
                 2701 
                 agactgtgga gactgctcct gagcccccag cttccagcag gagggacagt ctcaccattt 
               
               
                   
               
               
                 2761 
                 ccccagggca cgtggttgag tggggggaac gcccacttcc ctgggttaga ctgccagctc 
               
               
                   
               
               
                 2821 
                 ttcctagctg gagaggagcc ctgcctctcc gcccctgagc ccactgtgcg tggggctccc 
               
               
                   
               
               
                 2881 
                 gcctccaacc cctcgcccag tcccagcagc cagccaaaca cacagaaggg gactgccacc 
               
               
                   
               
               
                 2941 
                 tccccttgcc agctgctgag ccgcagagaa gtgacggttc ctacacagga caggggttcc 
               
               
                   
               
               
                 3001 
                 ttctgggcat tacatcgcat agaaatcaat aatttgtggt gatttggatc tgtgttttaa 
               
               
                   
               
               
                 3061 
                 tgagtttcac agtgtgattt tgattattaa ttgtgcaagc ttttcctaat aaacgtggag 
               
               
                   
               
               
                 3121 
                 aatcacaggc tgggctgggc actgctctca ccttggttcc tggggcatcc atggggtctc 
               
               
                   
               
               
                 3181 
                 tcacagacag gacccctgca gttcccctgg aagcagtgcc caggtggctg tggaatagga 
               
               
                   
               
               
                 3241 
                 acgctaaaaa aaaaaaaaaa aa 
               
            
           
         
       
     
     By “LGR5 polypeptide” is meant a protein or fragment thereof having at least 85% amino acid sequence identity to the sequence provided at NCBI Accession No. NP_003658.1 (isoform 1), NP_001264155.1 (isoform 2), or NP_001264156.1 (isoform 3) and having transmembrane signaling receptor activity or G-protein coupled receptor activity. The amino acid sequence provided at NCBI Accession No. NP_003658.1 is shown below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 19) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 mdtsrlgvll slpvllqlat ggssprsgvl lrgcpthchc epdgrmllrv dcsdlglsel 
                   
               
               
                   
               
               
                 61 
                 psnlsvftsy ldlsmnnisq llpnplpslr fleelrlagn altyipkgaf tglyslkvlm 
               
               
                   
               
               
                 121 
                 lqnnqlrhvp tealqnlrsl qslrldanhi syvppscfsg lhslrhlwld dnalteipvq 
               
               
                   
               
               
                 181 
                 afrslsalqa mtlalnkihh ipdyafgnls slvvlhlhnn rihslgkkcf dglhsletld 
               
               
                   
               
               
                 241 
                 lnynnldefp tairtlsnlk elgfhsnnir sipekafvgn pslitihfyd npiqfvgrsa 
               
               
                   
               
               
                 301 
                 fghlpelrtl tlngasqite fpdltgtanl esltltgaqi sslpqtvcnq lpnlqvldls 
               
               
                   
               
               
                 361 
                 ynlledlpsf svcqklqkid lrhneiyeik vdtfqqllsl rslnlawnki aiihpnafst 
               
               
                   
               
               
                 421 
                 lpslikldls snllssfpit glhglthlkl tgnhalqsli ssenfpelkv iempyayqcc 
               
               
                   
               
               
                 481 
                 afgvcenayk isnqwnkgdn ssmddlhkkd agmfqaqder dledflldfe edlkalhsvq 
               
               
                   
               
               
                 541 
                 cspspgpfkp cehlldgwli rigvwtiavl altcnalvts tvfrsplyis piklligvia 
               
               
                   
               
               
                 601 
                 avnmltgvss avlagvdaft fgsfarhgaw wengvgchvi gflsifases svflltlaal 
               
               
                   
               
               
                 661 
                 ergfsvkysa kfetkapfss lkviillcal laltmaavpl lggskygasp lclplpfgep 
               
               
                   
               
               
                 721 
                 stmgymvali llnslcflmm tiaytklycn ldkgdleniw dcsmvkhial llftncilnc 
               
               
                   
               
               
                 781 
                 pvaflsfssl inltfispev ikfillvvvp lpaclnplly ilfnphfked lvslrkqtyv 
               
               
                   
               
               
                 841 
                 wtrskhpslm sinsddvekq scdstqalvt ftsssitydl ppssvpspay pvteschlss 
               
               
                   
               
               
                 901 
                 vafvpcl 
               
            
           
         
       
     
     By “LGR5 polynucleotide” is meant a polynucleotide encoding a LGR5 polypeptide or fragment thereof. An exemplary LGR5 polynucleotide sequence is provided at NCBI Ref: NM_003667.3. The sequence provided at NCBI Ref: NM_003667.3 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 20) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 aaaaaacgag cgtgcaagca gagatgctgc tccacaccgc tcaggccgcg agcagcagca 
                   
               
               
                   
               
               
                 61 
                 aggcgcaccg ccactgtcgc cgctgcagcc agggctgctc cgaaggccgg cgtggcggca 
               
               
                   
               
               
                 121 
                 accggcacct ctgtccccgc cgcgcttctc ctcgccgccc acgccgtggg gtcaggaacg 
               
               
                   
               
               
                 181 
                 cggcgtctgg cgctgcagac gcccgctgag ttgcagaagc ccacggagcg gcgcccggcg 
               
               
                   
               
               
                 241 
                 cgccacggcc cgtagcagtc cggtgctgct ctccgcccgc gtccggctcg tggcccccta 
               
               
                   
               
               
                 301 
                 cttcgggcac catggacacc tcccggctcg gtgtgctcct gtccttgcct gtgctgctgc 
               
               
                   
               
               
                 361 
                 agctggcgac cgggggcagc tctcccaggt ctggtgtgtt gctgaggggc tgccccacac 
               
               
                   
               
               
                 421 
                 actgtcattg cgagcccgac ggcaggatgt tgctcagggt ggactgctcc gacctggggc 
               
               
                   
               
               
                 481 
                 tctcggagct gccttccaac ctcagcgtct tcacctccta cctagacctc agtatgaaca 
               
               
                   
               
               
                 541 
                 acatcagtca gctgctcccg aatcccctgc ccagtctccg cttcctggag gagttacgtc 
               
               
                   
               
               
                 601 
                 ttgcgggaaa cgctctgaca tacattccca agggagcatt cactggcctt tacagtctta 
               
               
                   
               
               
                 661 
                 aagttcttat gctgcagaat aatcagctaa gacacgtacc cacagaagct ctgcagaatt 
               
               
                   
               
               
                 721 
                 tgcgaagcct tcaatccctg cgtctggatg ctaaccacat cagctatgtg cccccaagct 
               
               
                   
               
               
                 781 
                 gtttcagtgg cctgcattcc ctgaggcacc tgtggctgga tgacaatgcg ttaacagaaa 
               
               
                   
               
               
                 841 
                 tccccgtcca ggcttttaga agtttatcgg cattgcaagc catgaccttg gccctgaaca 
               
               
                   
               
               
                 901 
                 aaatacacca cataccagac tatgcctttg gaaacctctc cagcttggta gttctacatc 
               
               
                   
               
               
                 961 
                 tccataacaa tagaatccac tccctgggaa agaaatgctt tgatgggctc cacagcctag 
               
               
                   
               
               
                 1021 
                 agactttaga tttaaattac aataaccttg atgaattccc cactgcaatt aggacactct 
               
               
                   
               
               
                 1081 
                 ccaaccttaa agaactagga tttcatagca acaatatcag gtcgatacct gagaaagcat 
               
               
                   
               
               
                 1141 
                 ttgtaggcaa cccttctctt attacaatac atttctatga caatcccatc cagtttgttg 
               
               
                   
               
               
                 1201 
                 ggagatctgc ttttcaacat ttacctgaac taagaacact gactctgaat ggtgcctcac 
               
               
                   
               
               
                 1261 
                 aaataactga atttcctgat ttaactggaa ctgcaaacct ggagagtctg actttaactg 
               
               
                   
               
               
                 1321 
                 gagcacagat ctcatctctt cctcaaaccg tctgcaatca gttacctaat ctccaagtgc 
               
               
                   
               
               
                 1381 
                 tagatctgtc ttacaaccta ttagaagatt tacccagttt ttcagtctgc caaaagcttc 
               
               
                   
               
               
                 1441 
                 agaaaattga cctaagacat aatgaaatct acgaaattaa agttgacact ttccagcagt 
               
               
                   
               
               
                 1501 
                 tgcttagcct ccgatcgctg aatttggctt ggaacaaaat tgctattatt caccccaatg 
               
               
                   
               
               
                 1561 
                 cattttccac tttgccatcc ctaataaagc tggacctatc gtccaacctc ctgtcgtctt 
               
               
                   
               
               
                 1621 
                 ttcctataac tgggttacat ggtttaactc acttaaaatt aacaggaaat catgccttac 
               
               
                   
               
               
                 1681 
                 agagcttgat atcatctgaa aactttccag aactcaaggt tatagaaatg ccttatgctt 
               
               
                   
               
               
                 1741 
                 accagtgctg tgcatttgga gtgtgtgaga atgcctataa gatttctaat caatggaata 
               
               
                   
               
               
                 1801 
                 aaggtgacaa cagcagtatg gacgaccttc ataagaaaga tgctggaatg tttcaggctc 
               
               
                   
               
               
                 1861 
                 aagatgaacg tgaccttgaa gatttcctgc ttgactttga ggaagacctg aaagcccttc 
               
               
                   
               
               
                 1921 
                 attcagtgca gtgttcacct tccccaggcc ccttcaaacc ctgtgaacac ctgcttgatg 
               
               
                   
               
               
                 1981 
                 gctggctgat cagaattgga gtgtggacca tagcagttct ggcacttact tgtaatgctt 
               
               
                   
               
               
                 2041 
                 tggtgacttc aacagttttc agatcccctc tgtacatttc ccccattaaa ctgttaattg 
               
               
                   
               
               
                 2101 
                 gggtcatcgc agcagtgaac atgctcacgg gagtctccag tgccgtgctg gctggtgtgg 
               
               
                   
               
               
                 2161 
                 atgcgttcac ttttggcagc tttgcacgac atggtgcctg gtgggagaat ggggttggtt 
               
               
                   
               
               
                 2221 
                 gccatgtcat tggttttttg tccatttttg cttcagaatc atctgttttc ctgcttactc 
               
               
                   
               
               
                 2281 
                 tggcagccct ggagcgtggg ttctctgtga aatattctgc aaaatttgaa acgaaagctc 
               
               
                   
               
               
                 2341 
                 cattttctag cctgaaagta atcattttgc tctgtgccct gctggccttg accatggccg 
               
               
                   
               
               
                 2401 
                 cagttcccct gctgggtggc agcaagtatg gcgcctcccc tctctgcctg cctttgcctt 
               
               
                   
               
               
                 2461 
                 ttggggagcc cagcaccatg ggctacatgg tcgctctcat cttgctcaat tccctttgct 
               
               
                   
               
               
                 2521 
                 tcctcatgat gaccattgcc tacaccaagc tctactgcaa tttggacaag ggagacctgg 
               
               
                   
               
               
                 2581 
                 agaatatttg ggactgctct atggtaaaac acattgccct gttgctcttc accaactgca 
               
               
                   
               
               
                 2641 
                 tcctaaactg ccctgtggct ttcttgtcct tctcctcttt aataaacctt acatttatca 
               
               
                   
               
               
                 2701 
                 gtcctgaagt aattaagttt atccttctgg tggtagtccc acttcctgca tgtctcaatc 
               
               
                   
               
               
                 2761 
                 cccttctcta catcttgttc aatcctcact ttaaggagga tctggtgagc ctgagaaagc 
               
               
                   
               
               
                 2821 
                 aaacctacgt ctggacaaga tcaaaacacc caagcttgat gtcaattaac tctgatgatg 
               
               
                   
               
               
                 2881 
                 tcgaaaaaca gtcctgtgac tcaactcaag ccttggtaac ctttaccagc tccagcatca 
               
               
                   
               
               
                 2941 
                 cttatgacct gcctcccagt tccgtgccat caccagctta tccagtgact gagagctgcc 
               
               
                   
               
               
                 3001 
                 atctttcctc tgtggcattt gtcccatgtc tctaattaat atgtgaagga aaatgttttc 
               
               
                   
               
               
                 3061 
                 aaaggttgag aacctgaaaa tgtgagattg agtatatcag agcagtaatt aataagaaga 
               
               
                   
               
               
                 3121 
                 gctgaggtga aactcggttt aaaaaccaaa aaagaatctc tcagttagta agaaaaggct 
               
               
                   
               
               
                 3181 
                 gaaaacctct tgatacttga gagtgaatat aagtctaaat gctgctttgt ataatttgtt 
               
               
                   
               
               
                 3241 
                 cagctaaggg atagatcgat cacactattt aagtgagccc agatcaaaaa agcagattga 
               
               
                   
               
               
                 3301 
                 aattttcttt agaaaagatt ctccatgatt tgaattgcat tctctttaaa ctcaccaatg 
               
               
                   
               
               
                 3361 
                 taatcatttt gggaggaggg agaacccact tgctttccaa atgggtttat ttaaacccac 
               
               
                   
               
               
                 3421 
                 aaactcaaga ggttgttggg ggaattagga aaataagggt tttcaatgac ctacattgct 
               
               
                   
               
               
                 3481 
                 aggtagaggc tgtgatccat gggatttcat tctaatgacc atgtgaagat gtttgagtcc 
               
               
                   
               
               
                 3541 
                 tcctttgcct ttcctcagaa agaatccttc taaggcacaa atcccttaga tggataatgt 
               
               
                   
               
               
                 3601 
                 aaggtattgt taactcactc atattgagat catttttaga gataccaggt tttatgtatc 
               
               
                   
               
               
                 3661 
                 agcactagat ggttccaccc tcatgggata aaactgctta caagtatttt gaaagaaaaa 
               
               
                   
               
               
                 3721 
                 ctgaccaaaa ttcttaaatt gttactaagg caatcatgca caggtgacgt atgtcttatc 
               
               
                   
               
               
                 3781 
                 tgatttgttt ttaactcctt ggtgcccaaa gctcagaagg gaattccact gccagcaatg 
               
               
                   
               
               
                 3841 
                 aacatacctg gaaaagaaag taagcaatct gggatttttt ttctgggtta gtaaagaatt 
               
               
                   
               
               
                 3901 
                 tttgcaataa gttttatcag ttgattcaaa ctgatgtgca tcttaatgat caaatgtgca 
               
               
                   
               
               
                 3961 
                 cattacataa attaagtcca ctgatacaac ttcttacaca tgtatctcta gtagctctgg 
               
               
                   
               
               
                 4021 
                 caaacccaat atctgacacc actttggact caagagactc agtaacgtat tatcctgttt 
               
               
                   
               
               
                 4081 
                 atttagcttg gttttagctg tgttctctct ggataaccca cttgatgtta ggaacattac 
               
               
                   
               
               
                 4141 
                 ttctctgctt attccatatt aatactgtgt taggtatttt aagaagcaag ttattaaata 
               
               
                   
               
               
                 4201 
                 agaaaagtca aagtattaat tcttaccttc tattatccta tattagcttc aatacatcca 
               
               
                   
               
               
                 4261 
                 aaccaaatgg ctgttaggta gatttatttt tatataagca tgtttatttt gatcagatgt 
               
               
                   
               
               
                 4321 
                 tttaacttgg atttgaaaaa atacatttat gagatgtttt ataagatgtg taaatataga 
               
               
                   
               
               
                 4381 
                 actgtattta ttactatagt aaaggttcag taacattaag gaccatgata atgataataa 
               
               
                   
               
               
                 4441 
                 accttgtaca gtggcatatt ctttgattta tattgtgttt ctctgcccat tttctttaaa 
               
               
                   
               
               
                 4501 
                 ttcattaact gtatatatgt aaatatatag tacttgtaaa tagattccaa atttgctttt 
               
               
                   
               
               
                 4561 
                 ctattgggta aaaaataaat ttgtaataaa atgtgtgact atgaaacaaa aaaaaaaaaa 
               
               
                   
               
               
                 4621 
                 aaaaa 
               
            
           
         
       
     
     By “LDHA polypeptide” or “lactate dehydrogenase A polypeptide” is meant a protein or fragment thereof having at least 85% amino acid sequence identity to the sequence provided at NCBI Accession No. NP_005557.1 (isoform 1), NP_001128711.1 (isoform 2), NP_001158886.1 (isoform 3), NP_001158887.1 (isoform 4), or NP_001158888.1 (isoform 5) and having dehydrogenase activity. The amino acid sequence provided at NCBI Accession No. NP_005557.1 is shown below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 21) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 matlkdqliy nllkeeqtpq nkitvvgvga vgmacaisil mkdladelal vdviedklkg 
                   
               
               
                   
               
               
                 61 
                 emmdlqhgsl flrtpkivsg kdynvtansk lviitagarq qegesrlnlv qrnvnifkfi 
               
               
                   
               
               
                 121 
                 ipnvvkyspn ckllivsnpv diltyvawki sgfpknrvig sgcnldsarf rylmgerlgv 
               
               
                   
               
               
                 181 
                 hplschgwvl gehgdssvpv wsgmnvagvs lktlhpdlgt dkdkeqwkev hkqvvesaye 
               
               
                   
               
               
                 241 
                 viklkgytsw aiglsvadla esimknlrrv hpvstmikgl ygikddvfls vpcilgqngi 
               
               
                   
               
               
                 301 
                 sdlvkvtlts eeearlkksa dtlwgiqkel qf 
               
            
           
         
       
     
     By “LDHA polynucleotide” or “lactate dehydrogenase A polynucleotide” is meant a polynucleotide encoding a LDHA polypeptide or fragment thereof. An exemplary LDHA polynucleotide sequence is provided at NCBI Ref: NM_005566.3. The sequence provided at NCBI Ref: NM_005566.3 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 22) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 gtctgccggt cggttgtctg gctgcgcgcg ccacccgggc ctctccagtg ccccgcctgg 
                   
               
               
                   
               
               
                 61 
                 ctcggcatcc acccccagcc cgactcacac gtgggttccc gcacgtccgc cggccccccc 
               
               
                   
               
               
                 121 
                 cgctgacgtc agcatagctg ttccacttaa ggcccctccc gcgcccagct cagagtgctg 
               
               
                   
               
               
                 181 
                 cagccgctgc cgccgattcc ggatctcatt gccacgcgcc cccgacgacc gcccgacgtg 
               
               
                   
               
               
                 241 
                 cattcccgat tccttttggt tccaagtcca atatggcaac tctaaaggat cagctgattt 
               
               
                   
               
               
                 301 
                 ataatcttct aaaggaagaa cagacccccc agaataagat tacagttgtt ggggttggtg 
               
               
                   
               
               
                 361 
                 ctgttggcat ggcctgtgcc atcagtatct taatgaagga cttggcagat gaacttgctc 
               
               
                   
               
               
                 421 
                 ttgttgatgt catcgaagac aaattgaagg gagagatgat ggatctccaa catggcagcc 
               
               
                   
               
               
                 481 
                 ttttccttag aacaccaaag attgtctctg gcaaagacta taatgtaact gcaaactcca 
               
               
                   
               
               
                 541 
                 agctggtcat tatcacggct ggggcacgtc agcaagaggg agaaagccgt cttaatttgg 
               
               
                   
               
               
                 601 
                 tccagcgtaa cgtgaacatc tttaaattca tcattcctaa tgttgtaaaa tacagcccga 
               
               
                   
               
               
                 661 
                 actgcaagtt gcttattgtt tcaaatccag tggatatctt gacctacgtg gcttggaaga 
               
               
                   
               
               
                 721 
                 taagtggttt tcccaaaaac cgtgttattg gaagcggttg caatctggat tcagcccgat 
               
               
                   
               
               
                 781 
                 tccgttacct aatgggggaa aggctgggag ttcacccatt aagctgtcat gggtgggtcc 
               
               
                   
               
               
                 841 
                 ttggggaaca tggagattcc agtgtgcctg tatggagtgg aatgaatgtt gctggtgtct 
               
               
                   
               
               
                 901 
                 ctctgaagac tctgcaccca gatttaggga ctgataaaga taaggaacag tggaaagagg 
               
               
                   
               
               
                 961 
                 ttcacaagca ggtggttgag agtgcttatg aggtgatcaa actcaaaggc tacacatcct 
               
               
                   
               
               
                 1021 
                 gggctattgg actctctgta gcagatttgg cagagagtat aatgaagaat cttaggcggg 
               
               
                   
               
               
                 1081 
                 tgcacccagt ttccaccatg attaagggtc tttacggaat aaaggatgat gtcttcctta 
               
               
                   
               
               
                 1141 
                 gtgttccttg cattttggga cagaatggaa tctcagacct tgtgaaggtg actctgactt 
               
               
                   
               
               
                 1201 
                 ctgaggaaga ggcccgtttg aagaagagtg cagatacact ttgggggatc caaaaggagc 
               
               
                   
               
               
                 1261 
                 tgcaatttta aagtcttctg atgtcatatc atttcactgt ctaggctaca acaggattct 
               
               
                   
               
               
                 1321 
                 aggtggaggt tgtgcatgtt gtccttttta tctgatctgt gattaaagca gtaatatttt 
               
               
                   
               
               
                 1381 
                 aagatggact gggaaaaaca tcaactcctg aagttagaaa taagaatggt ttgtaaaatc 
               
               
                   
               
               
                 1441 
                 cacagctata tcctgatgct ggatggtatt aatcttgtgt agtcttcaac tggttagtgt 
               
               
                   
               
               
                 1501 
                 gaaatagttc tgccacctct gacgcaccac tgccaatgct gtacgtactg catttgcccc 
               
               
                   
               
               
                 1561 
                 ttgagccagg tggatgttta ccgtgtgtta tataacttcc tggctccttc actgaacatg 
               
               
                   
               
               
                 1621 
                 cctagtccaa cattttttcc cagtgagtca catcctggga tccagtgtat aaatccaata 
               
               
                   
               
               
                 1681 
                 tcatgtcttg tgcataattc ttccaaagga tcttattttg tgaactatat cagtagtgta 
               
               
                   
               
               
                 1741 
                 cattaccata taatgtaaaa agatctacat acaaacaatg caaccaacta tccaagtgtt 
               
               
                   
               
               
                 1801 
                 ataccaacta aaacccccaa taaaccttga acagtgacta ctttggttaa ttcattatat 
               
               
                   
               
               
                 1861 
                 taagatataa agtcataaag ctgctagtta ttatattaat ttggaaatat taggctattc 
               
               
                   
               
               
                 1921 
                 ttgggcaacc ctgcaacgat tttttctaac agggatatta ttgactaata gcagaggatg 
               
               
                   
               
               
                 1981 
                 taatagtcaa ctgagttgta ttggtaccac ttccattgta agtcccaaag tattatatat 
               
               
                   
               
               
                 2041 
                 ttgataataa tgctaatcat aattggaaag taacattcta tatgtaaatg taaaatttat 
               
               
                   
               
               
                 2101 
                 ttgccaactg aatataggca atgatagtgt gtcactatag ggaacacaga tttttgagat 
               
               
                   
               
               
                 2161 
                 cttgtcctct ggaagctggt aacaattaaa aacaatctta aggcagggaa aaaaaaaaaa 
               
               
                   
               
               
                 2221 
                 aaaaaa 
               
            
           
         
       
     
     By “MAFA polypeptide” is meant a protein or fragment thereof having at least 85% amino acid sequence identity to the sequence provided at NCBI Accession No. NP_963883.2 and having transcription factor activity. The amino acid sequence provided at NCBI Accession No. NP_963883.2 is shown below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 23) 
                   
               
            
           
           
               
               
               
            
               
                   1 
                 maaelamgae lpssplaiey vndfdlmkfe vkkeppeaer fchrlppgsl sstplstpcs 
                   
               
               
                   
               
               
                  61 
                 svpsspsfca pspgtggggg agggggssqa ggapgppsgg pgavggtsgk paledlywms 
               
               
                   
               
               
                 121 
                 gyqhhlnpea lnltpedave aligsghhga hhgahhpaaa aayeafrgpg faggggaddm 
               
               
                   
               
               
                 181 
                 gaghhhgahh aahhhhaahh hhhhhhhhgg aghgggaghh vrleerfsdd qlvsmsvrel 
               
               
                   
               
               
                 241 
                 nrqlrgfske evirlkqkrr tlknrgyaqs crfkrvqqrh ilesekcqlq sqveqlklev 
               
               
                   
               
               
                 301 
                 grlakerdly kekyeklagr ggpgsaggag fprepsppqa gpggakgtad ffl 
               
            
           
         
       
     
     By “MAFA polynucleotide” is meant a polynucleotide encoding a MAFA polypeptide or fragment thereof. An exemplary MAFA polynucleotide sequence is provided at NCBI Ref: NM_201589.3. The sequence provided at NCBI Ref: NM_201589.3 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 24) 
                   
               
            
           
           
               
               
               
            
               
                   1 
                 gcgcggccgg gcgcgggccc cgggcgatgg ccgcggagct ggcgatgggc gccgagctgc 
                   
               
               
                   
               
               
                  61 
                 ccagcagccc gctggccatc gagtacgtca acgacttcga cctgatgaag ttcgaggtga 
               
               
                   
               
               
                  121 
                 agaaggagcc tcccgaggcc gagcgcttct gccaccgcct gccgccaggc tcgctgtcct 
               
               
                   
               
               
                  181 
                 cgacgccgct cagcacgccc tgctcctccg tgccctcctc gcccagcttc tgcgcgccca 
               
               
                   
               
               
                  241 
                 gcccgggcac cggcggcggc ggcggcgcgg ggggcggcgg cggctcgtct caggccgggg 
               
               
                   
               
               
                  301 
                 gcgcccccgg gccgccgagc gggggccccg gcgccgtcgg gggcacctcg gggaagccgg 
               
               
                   
               
               
                  361 
                 cgctggagga tctgtactgg atgagcggct accagcatca cctcaacccc gaggcgctca 
               
               
                   
               
               
                  421 
                 acctgacgcc cgaggacgcg gtggaggcgc tcatcggcag cggccaccac ggcgcgcacc 
               
               
                   
               
               
                  481 
                 acggcgcgca ccacccggcg gccgccgcag cctacgaggc tttccgcggc ccgggcttcg 
               
               
                   
               
               
                  541 
                 cgggcggcgg cggagcggac gacatgggcg ccggccacca ccacggcgcg caccacgccg 
               
               
                   
               
               
                  601 
                 cccaccatca ccacgccgcc caccaccacc accaccacca ccaccaccat ggcggcgcgg 
               
               
                   
               
               
                  661 
                 gacacggcgg tggcgcgggc caccacgtgc gcctggagga gcgcttctcc gacgaccagc 
               
               
                   
               
               
                  721 
                 tggtgtccat gtcggtgcgc gagctgaacc ggcagctccg cggcttcagc aaggaggagg 
               
               
                   
               
               
                  781 
                 tcatccggct caagcagaag cggcgcacgc tcaagaaccg cggctacgcg cagtcctgcc 
               
               
                   
               
               
                  841 
                 gcttcaagcg ggtgcagcag cggcacattc tggagagcga gaagtgccaa ctccagagcc 
               
               
                   
               
               
                  901 
                 aggtggagca gctgaagctg gaggtggggc gcctggccaa agagcgggac ctgtacaagg 
               
               
                   
               
               
                  961 
                 agaaatacga gaagctggcg ggccggggcg gccccgggag cgcgggcggg gccggtttcc 
               
               
                   
               
               
                 1021 
                 cgcgggagcc ttcgccgccg caggccggtc ccggcggggc caagggcacg gccgacttct 
               
               
                   
               
               
                 1081 
                 tcctgtaggc gccggacccc gagcccgcgc cgccgtcgcc ggggacaagt tcgcgcaggc 
               
               
                   
               
               
                 1141 
                 ctctcggggc ctcggctcgg actccgcggt acaggacgtg gacaccaggc ccggcccggc 
               
               
                   
               
               
                 1201 
                 cgtgctggcc ccggtgccaa gtctgcgggc gcggggctgg aggccccttc gctcccggtc 
               
               
                   
               
               
                 1261 
                 cccgttcgcg cgcgtcggcc cgggtcgccg tcctgaggtt gagcggagaa cggtgatttc 
               
               
                   
               
               
                 1321 
                 taaggaaact tgagccaggt ctaacttctt tccaagcgtc cgcttgtaca tacgttgaac 
               
               
                   
               
               
                 1381 
                 gtggttctcc gttcccacct tcgccctgcc agcctagagg gaccgcgctg ccgtcccttc 
               
               
                   
               
               
                 1441 
                 ccgggtggcc cctgcctgcc cccgccctcc ttcgttctct tctcagcctc cctttccttg 
               
               
                   
               
               
                 1501 
                 ccttttttaa cttcccctcc ccgttttaaa atcggtctta ttttcgaagt atttataatt 
               
               
                   
               
               
                 1561 
                 attatgcttg gtgattagaa aagaaaacct tggaggaagc cccttctttc cccagccggg 
               
               
                   
               
               
                 1621 
                 gtccgccctc agtcgcgagt cacagcatga gtcgctcgcc aggaggggcc cggcccctgc 
               
               
                   
               
               
                 1681 
                 ctgccccctc cccgcttgcc cccgaccctg ctaccggcgt tccttggagg tcgaagccag 
               
               
                   
               
               
                 1741 
                 ggacgtcacc cgtgctgtgt ccaggcctgc tgtcctacta tgctcaaccg ggggtggggg 
               
               
                   
               
               
                 1801 
                 gaggggggtg agtcctgtgc tcagtcgggt gggggctggc ccggatcccg agctgctgtc 
               
               
                   
               
               
                 1861 
                 tctctatgca ccagaacata tctgtaactc ctggggaaat acatcttgtt ttaaccttca 
               
               
                   
               
               
                 1921 
                 agagaagtga aagaaaaaag taatgcacag tatttctagc agaaaatttt tttttttaag 
               
               
                   
               
               
                 1981 
                 aggaggcttg ggccagagcc ttctggcatg gggcgggtgg agaaagtgtt tttattttaa 
               
               
                   
               
               
                 2041 
                 tttaaattgt gtttcgtttt gtttgtggaa tctttcttta atgcttcgtc gctctttgga 
               
               
                   
               
               
                 2101 
                 ctagccggga gagagggcga ggaggcgggt gctccaggcc ctgtaggctg ggccaggcgc 
               
               
                   
               
               
                 2161 
                 ctgggggatc tgcccgtttt cggaggccct caggggccat cagtgggatt ccagccgctc 
               
               
                   
               
               
                 2221 
                 cacacccctc ccctgagcac tcggagtgga aggcgcgccg actcgttgaa agttttgttg 
               
               
                   
               
               
                 2281 
                 tgtagttggt tttcgttgag ttcttttttc atttgctacg aaactgagaa aaagaaaaaa 
               
               
                   
               
               
                 2341 
                 atacacaaaa taaatctgtt cagatccaag tca 
               
            
           
         
       
     
     As used herein, a “marker” is meant any protein or polynucleotide having an alteration in expression level or activity that is associated with a disease or disorder or that is associated with a particular cell type. In some embodiments, a marker for a beta cell is Pdx1, MafA, Pax4, Pax6, NeuroD1, Nkx6-1, Gata6, or Foxa2. In some embodiments, a marker for a hepatocyte is AFP, ALB, or Cyp3a7. In some other embodiments, a marker for a cardiomyocyte is hMlc2a, hNkx2-5, alphaMHC or KCNQ1. In still other embodiments, a marker for a small intestine cell is CDX2, Muc2, or Lgr5. 
     By “alphaMHC polypeptide” or “myosin heavy chain (MHC) alpha polypeptide” is meant a protein or fragment thereof having at least 85% amino acid sequence identity to the sequence provided at NCBI Accession No. NP_002462.2 and having actin binding activity. The amino acid sequence provided at NCBI Accession No. NP_002462.2 is shown below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 25) 
                   
               
            
           
           
               
               
               
            
               
                   1 
                 mtdaqmadfg aaaqylrkse kerleaqtrp fdirtecfvp ddkeefvkak ilsreggkvi 
                   
               
               
                   
               
               
                  61 
                 aetengktvt vkedqvlqqn ppkfdkiedm amltflhepa vlfnlkerya awmiytysgl 
               
               
                   
               
               
                  121 
                 fcvtvnpykw lpvynaevva ayrgkkrsea pphifsisdn ayqymltdre nqsilitges 
               
               
                   
               
               
                  181 
                 gagktvntkr viqyfasiaa igdrgkkdna nankgtledq iiqanpalea fgnaktvrnd 
               
               
                   
               
               
                  241 
                 nssrfgkfir ihfgatgkla sadietylle ksrvifqlka ernyhifyqi lsnkkpelld 
               
               
                   
               
               
                  301 
                 mllvtnnpyd yafvsqgevs vasiddseel matdsafdvl gftseekagv ykltgaimhy 
               
               
                   
               
               
                  361 
                 gnmkfkqkqr eeqaepdgte dadksaylmg lnsadllkgl chprvkvgne yvtkggsvgq 
               
               
                   
               
               
                  421 
                 vyysigalak avyekmfnwm vtrinatlet kqprgyfigv ldiagfeifd fnsfeqlcin 
               
               
                   
               
               
                  481 
                 ftneklqqff nhhmfvleqe eykkegiewt fidfgmdlqa cidliekpmg imsileeecm 
               
               
                   
               
               
                  541 
                 fpkatdmtfk aklydnhlgk snnfqkprni kgkqeahfsl ihyagtvdyn ilgwleknkd 
               
               
                   
               
               
                  601 
                 pinetvvaly qksslklmat lfssyatadt gdsgkskggk kkgssfqtvs alhrenlnkl 
               
               
                   
               
               
                  661 
                 mtnlrtthph fvrciipner kapgvmdnpl vmhqlrongv legiricrkg fpnrilygdf 
               
               
                   
               
               
                  721 
                 rqryrilnpv aipegqfids rkgtekllss ldidhnqykf ghtkvffkag llglleemrd 
               
               
                   
               
               
                  781 
                 erlsriitrm gagargqlmr iefkkiverr dallviqwni rafmgvknwp wmklyfkikp 
               
               
                   
               
               
                  841 
                 llksaeteke matmkeefgr iketleksea rrkeleekmv sllgekndlq lqvgaegdnl 
               
               
                   
               
               
                  901 
                 ndaeercdql iknkiqleak vkemnerled eeemnaelta kkrkledecs elkkdiddle 
               
               
                   
               
               
                  961 
                 ltlakvekek hatenkvknl teemagldei iakltkekka lqeahqqald dlqveedkvn 
               
               
                   
               
               
                 1021 
                 slskskvkle qqvddlegsl egekkvrmdl erakrklegd lkltqesimd lendklqlee 
               
               
                   
               
               
                 1081 
                 klkkkefdin qqnskiedeq vlalqlqkkl kenqarieel eeeleaerta rakveklrsd 
               
               
                   
               
               
                 1141 
                 lsreleeise rleeaggats vqiemnkkre aefqkmrrdl eeatlqheat aaalrkkhad 
               
               
                   
               
               
                 1201 
                 svaelgeqid nlqrvkqkle keksefklel ddvtsnmeqi ikakanlekv srtledqane 
               
               
                   
               
               
                 1261 
                 yrvkleeaqr slndfttqra klqtengela rqleekeali sqltrgklsy tqqmedlkrq 
               
               
                   
               
               
                 1321 
                 leeegkakna lahalqsarh dcdllregye eeteakaelq rvlskansev aqwrtkyetd 
               
               
                   
               
               
                 1381 
                 aiqrteelee akkklaqrlq daeeaveavn akcsslektk hrlgneiedl mvdversnaa 
               
               
                   
               
               
                 1441 
                 aaaldkkqrn fdkilaewkq kyeesqsele ssqkearsls telfklknay eeslehletf 
               
               
                   
               
               
                 1501 
                 krenknlqee isdlteqlge ggknvhelek vrkqlevekl elqsaleeae asleheegki 
               
               
                   
               
               
                 1561 
                 lraglefnqi kaeierklae kdeemeqakr nhqrvvdslq tsldaetrsr nevlrvkkkm 
               
               
                   
               
               
                 1621 
                 egdlnemeiq lshanrmaae aqkqvkslqs llkdtgiqld davranddlk eniaiverrn 
               
               
                   
               
               
                 1681 
                 nllqaeleel ravveqters rklaeqelie tservqllhs qntslinqkk kmesdltqlq 
               
               
                   
               
               
                 1741 
                 seveeavqec rnaeekakka itdaammaee lkkeqdtsah lermkknmeq tikdlqhrld 
               
               
                   
               
               
                 1801 
                 eaeqialkgg kkqlqklear vrelegelea eqkrnaesvk gmrkserrik eltyqteedk 
               
               
                   
               
               
                 1861 
                 knllrlqdlv dklqlkvkay krqaeeaeeq antnlskfrk vqheldeaee radiaesqvn 
               
               
                   
               
               
                 1921 
                 klraksrdig akqkmhdee 
               
            
           
         
       
     
     By “alphaMHC polynucleotide” is meant a polynucleotide encoding a alphaMHC polypeptide or fragment thereof. An exemplary alphaMHC polynucleotide sequence is provided at NCBI Ref: NM_002471.3. The sequence provided at NCBI Ref: NM_002471.3 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 26) 
                   
               
            
           
           
               
               
               
            
               
                   1 
                 agatagagag actcctgcgg cccagattct tcaggattct ccgtgaaggg ataaccaggg 
                   
               
               
                   
               
               
                  61 
                 gaagcaccaa gatgaccgat gcccagatgg ctgactttgg ggcagcggcc cagtacctcc 
               
               
                   
               
               
                  121 
                 gcaagtcaga gaaggagcgt ctagaggccc agacccggcc ctttgacatt cgcactgagt 
               
               
                   
               
               
                  181 
                 gcttcgtgcc cgatgacaag gaagagtttg tcaaagccaa gattttgtcc cgggagggag 
               
               
                   
               
               
                  241 
                 gcaaggtcat tgctgaaacc gagaatggga agacggtgac tgtgaaggag gaccaggtgt 
               
               
                   
               
               
                  301 
                 tgcagcagaa cccacccaag ttcgacaaga ttgaggacat ggccatgctg accttcctgc 
               
               
                   
               
               
                  361 
                 acgagcccgc ggtgcttttc aacctcaagg agcgctacgc ggcctggatg atatatacct 
               
               
                   
               
               
                  421 
                 actcgggcct cttctgtgtc actgtcaacc cctacaagtg gctgccggtg tacaatgccg 
               
               
                   
               
               
                  481 
                 aggtggtggc cgcctaccgg ggcaagaaga ggagtgaggc cccgccccac atcttctcca 
               
               
                   
               
               
                  541 
                 tctccgacaa cgcctatcag tacatgctga cagatcggga gaaccagtcc atcctcatca 
               
               
                   
               
               
                  601 
                 cgggagaatc cggggcgggg aagactgtga acaccaagcg tgtcatccag tactttgcca 
               
               
                   
               
               
                  661 
                 gcattgcagc cataggtgac cgtggcaaga aggacaatgc caatgcgaac aagggcaccc 
               
               
                   
               
               
                  721 
                 tggaggacca gatcatccag gccaaccccg ctctggaggc cttcggcaat gccaagactg 
               
               
                   
               
               
                  781 
                 tccggaacga caactcctcc cgctttggga aattcattag gatccacttt ggggccactg 
               
               
                   
               
               
                  841 
                 gaaagctggc ttctgcagac atagagacct acctgctgga gaagtcccgg gtgatcttcc 
               
               
                   
               
               
                  901 
                 agctgaaagc tgagagaaac taccacatct tctaccagat tctgtccaac aagaagccgg 
               
               
                   
               
               
                  961 
                 agttgctgga catgctgctg gtcaccaaca atccctacga ctacgccttc gtgtctcagg 
               
               
                   
               
               
                 1021 
                 gagaggtgtc cgtggcctcc attgatgact ccgaggagct catggccacc gatagtgcct 
               
               
                   
               
               
                 1081 
                 ttgacgtgct gggcttcact tcagaggaga aagctggcgt ctacaagctg acgggagcca 
               
               
                   
               
               
                 1141 
                 tcatgcacta cgggaacatg aagttcaagc agaagcagcg ggaggagcag gcggagccag 
               
               
                   
               
               
                 1201 
                 acggcaccga agatgctgac aagtcggcct acctcatggg gctgaactca gctgacctgc 
               
               
                   
               
               
                 1261 
                 tcaaggggct gtgccaccct cgggtgaaag tgggcaacga gtatgtcacc aaggggcaga 
               
               
                   
               
               
                 1321 
                 gcgtgcagca ggtgtactac tccatcgggg ctctggccaa ggcagtgtat gagaagatgt 
               
               
                   
               
               
                 1381 
                 tcaactggat ggtgacgcgc atcaacgcca ccctggagac caagcagcca cgccagtact 
               
               
                   
               
               
                 1441 
                 tcataggagt cctggacatc gctggcttcg agatcttcga cttcaacagc tttgagcagc 
               
               
                   
               
               
                 1501 
                 tctgcatcaa cttcaccaac gagaagctgc agcagttctt caaccaccac atgttcgtgc 
               
               
                   
               
               
                 1561 
                 tggagcagga ggagtacaag aaggagggca ttgagtggac attcattgac tttggcatgg 
               
               
                   
               
               
                 1621 
                 acctgcaggc ctgcattgac ctcatcgaga agcccatggg catcatgtcc atcctggagg 
               
               
                   
               
               
                 1681 
                 aggagtgcat gttccccaag gccactgaca tgaccttcaa ggccaagctg tacgacaacc 
               
               
                   
               
               
                 1741 
                 acctgggcaa gtccaacaat ttccagaagc cacgcaacat caaggggaag caggaagccc 
               
               
                   
               
               
                 1801 
                 acttctccct gatccactac gccggcactg tggactacaa catcctgggc tggctggaaa 
               
               
                   
               
               
                 1861 
                 aaaacaagga tcctctcaac gagactgttg tggccctgta ccagaagtcc tccctcaagc 
               
               
                   
               
               
                 1921 
                 tcatggccac tctcttctcc tcctacgcaa ctgccgatac tggggacagt ggtaaaagca 
               
               
                   
               
               
                 1981 
                 aaggaggcaa gaaaaagggc tcatccttcc agacggtgtc ggctctccac cgggaaaatc 
               
               
                   
               
               
                 2041 
                 tcaacaagct aatgaccaac ctgaggacca cccatcctca ctttgtgcgt tgcatcatcc 
               
               
                   
               
               
                 2101 
                 ccaatgagcg gaaggctcca ggggtgatgg acaaccccct ggtcatgcac cagctgcgct 
               
               
                   
               
               
                 2161 
                 gcaatggcgt gctggagggc atccgcatct gcaggaaggg cttccccaac cgcatcctct 
               
               
                   
               
               
                 2221 
                 acggggactt ccggcagagg tatcgcatcc tgaacccagt ggccatccct gagggacagt 
               
               
                   
               
               
                 2281 
                 tcattgatag caggaagggg acagagaagc tgctcagctc tctggacatt gatcacaacc 
               
               
                   
               
               
                 2341 
                 agtacaagtt tggccacacc aaggtgttct tcaaggcagg gctgcttggg ctgctggagg 
               
               
                   
               
               
                 2401 
                 agatgcggga tgagaggctg agccgcatca tcacgcgcat gcaggcccaa gcccggggcc 
               
               
                   
               
               
                 2461 
                 agctcatgcg cattgagttc aagaagatag tggaacgcag ggatgccctg ctggtaatcc 
               
               
                   
               
               
                 2521 
                 agtggaacat tcgggccttc atgggggtca agaattggcc ctggatgaag ctctacttca 
               
               
                   
               
               
                 2581 
                 agatcaagcc gctgctgaag agcgcagaga cggagaagga gatggccacc atgaaggaag 
               
               
                   
               
               
                 2641 
                 agttcgggcg catcaaagag acgctggaga agtccgaggc tcgccgcaag gagctggagg 
               
               
                   
               
               
                 2701 
                 agaagatggt gtccctgctg caggagaaga atgacctgca gctccaagtg caggcggaac 
               
               
                   
               
               
                 2761 
                 aagacaacct caatgatgct gaggagcgct gcgaccagct gatcaaaaac aagattcagc 
               
               
                   
               
               
                 2821 
                 tggaggccaa agtaaaggag atgaatgaga ggctggagga tgaggaggag atgaacgcgg 
               
               
                   
               
               
                 2881 
                 agctcactgc caagaagcgc aagctggaag acgagtgctc agagctcaag aaggacattg 
               
               
                   
               
               
                 2941 
                 atgacctgga gctgacactg gccaaggtgg agaaggagaa gcatgcaaca gagaacaagg 
               
               
                   
               
               
                 3001 
                 tgaagaacct aacagaggag atggctgggc tggatgaaat catcgctaag ctgaccaagg 
               
               
                   
               
               
                 3061 
                 agaagaaagc tctacaagag gcccatcagc aggccctgga tgaccttcag gttgaggaag 
               
               
                   
               
               
                 3121 
                 acaaggtcaa cagcctgtcc aagtctaagg tcaagctgga gcagcaggtg gatgatctgg 
               
               
                   
               
               
                 3181 
                 agggatccct agagcaagag aagaaggtgc gcatggacct ggagcgagca aagcggaaac 
               
               
                   
               
               
                 3241 
                 tggagggcga cctgaagctg acccaggaga gcatcatgga cctggaaaat gataaactgc 
               
               
                   
               
               
                 3301 
                 agctggaaga aaagcttaag aagaaggagt ttgacattaa tcagcagaac agtaagattg 
               
               
                   
               
               
                 3361 
                 aggatgagca ggtgctggcc cttcaactac agaagaaact gaaggaaaac caggcacgca 
               
               
                   
               
               
                 3421 
                 tcgaggagct ggaggaggag ctggaggccg agcgcaccgc cagggctaag gtggagaagc 
               
               
                   
               
               
                 3481 
                 tgcgctcaga cctgtctcgg gagctggagg agatcagcga gcggctggaa gaggccggcg 
               
               
                   
               
               
                 3541 
                 gggccacgtc cgtgcagatc gagatgaaca agaagcgcga ggccgagttc cagaagatgc 
               
               
                   
               
               
                 3601 
                 ggcgggacct ggaggaggcc acgctgcagc acgaggccac tgccgcggcc ctgcgcaaga 
               
               
                   
               
               
                 3661 
                 agcacgccga cagcgtggcc gagctgggcg agcagatcga caacctgcag cgggtgaagc 
               
               
                   
               
               
                 3721 
                 agaagctgga gaaggagaag agcgagttca agctggagct ggatgacgtc acctccaaca 
               
               
                   
               
               
                 3781 
                 tggagcagat catcaaggcc aaggcaaacc tggagaaagt gtctcggacg ctggaggacc 
               
               
                   
               
               
                 3841 
                 aggccaatga gtaccgcgtg aagctagaag aggcccaacg ctccctcaat gatttcacca 
               
               
                   
               
               
                 3901 
                 cccagcgagc caagctgcag accgagaatg gagagttggc ccggcagcta gaggaaaagg 
               
               
                   
               
               
                 3961 
                 aggcgctaat ctcgcagctg acccggggga agctctctta tacccagcaa atggaggacc 
               
               
                   
               
               
                 4021 
                 tcaaaaggca gctggaggag gagggcaagg cgaagaacgc cctggcccat gcactgcagt 
               
               
                   
               
               
                 4081 
                 cggcccggca tgactgcgac ctgctgcggg agcagtacga ggaggagaca gaggccaagg 
               
               
                   
               
               
                 4141 
                 ccgagctgca gcgcgtcctg tccaaggcca actcggaggt ggcccagtgg aggaccaagt 
               
               
                   
               
               
                 4201 
                 atgagacgga cgccattcag cggactgagg agctcgaaga ggccaaaaag aagctggccc 
               
               
                   
               
               
                 4261 
                 agcggctgca ggatgccgag gaggccgtgg aggctgttaa tgccaagtgc tcctcactgg 
               
               
                   
               
               
                 4321 
                 agaagaccaa gcaccggcta cagaatgaga tagaggactt gatggtggac gtagagcgct 
               
               
                   
               
               
                 4381 
                 ccaatgctgc tgctgcagcc ctggacaaga agcagagaaa ctttgacaag atcctggccg 
               
               
                   
               
               
                 4441 
                 agtggaagca gaagtatgag gagtcgcagt ctgagctgga gtcctcacag aaggaggctc 
               
               
                   
               
               
                 4501 
                 gctccctcag cacagagctc ttcaagctca agaacgccta cgaggagtcc ctggagcacc 
               
               
                   
               
               
                 4561 
                 tagagacctt caagcgggag aacaagaacc ttcaggagga aatctcggac cttactgagc 
               
               
                   
               
               
                 4621 
                 agctaggaga aggaggaaag aatgtgcatg agctggagaa ggtccgcaaa cagctggagg 
               
               
                   
               
               
                 4681 
                 tggagaagct ggagctgcag tcagccctgg aggaggcaga ggcctccctg gagcacgagg 
               
               
                   
               
               
                 4741 
                 agggcaagat cctccgggcc cagctagagt tcaaccagat caaggcagag atcgagcgga 
               
               
                   
               
               
                 4801 
                 agctggcaga gaaggacgag gagatggaac aggccaagcg caaccaccag cgggtggtgg 
               
               
                   
               
               
                 4861 
                 actcgctgca gacctccctg gatgcagaga cacgcagccg caacgaggtc ctgagggtga 
               
               
                   
               
               
                 4921 
                 agaagaagat ggaaggagac ctcaatgaga tggagatcca gctcagccac gccaaccgca 
               
               
                   
               
               
                 4981 
                 tggctgccga ggcccagaag caagtcaaga gcctccagag cttgctgaag gacacccaga 
               
               
                   
               
               
                 5041 
                 tccagctgga cgatgcggtc cgtgccaacg acgacctgaa ggagaacatc gccatcgtgg 
               
               
                   
               
               
                 5101 
                 agcggcgcaa caacctgctg caggctgagc tggaggagct gcgtgccgtg gtggagcaga 
               
               
                   
               
               
                 5161 
                 cagagcggtc ccggaagctg gcggagcagg agctgattga gaccagcgag cgggtgcagc 
               
               
                   
               
               
                 5221 
                 tgctgcattc ccagaacacc agcctcatca accagaagaa gaagatggag tcggatctga 
               
               
                   
               
               
                 5281 
                 cccagctcca gtcggaagtg gaggaggcag tgcaggagtg cagaaacgcc gaggagaagg 
               
               
                   
               
               
                 5341 
                 ccaagaaggc catcacggat gccgccatga tggcagagga gctgaagaag gagcaggaca 
               
               
                   
               
               
                 5401 
                 ccagcgccca cctggagcgc atgaagaaga acatggagca gaccattaag gacctgcagc 
               
               
                   
               
               
                 5461 
                 accggctgga cgaggccgag cagatcgccc tcaagggagg caagaagcag ctgcagaagc 
               
               
                   
               
               
                 5521 
                 tggaagcgcg ggtgcgggag ctggagggtg agctggaggc cgagcagaag cgcaacgcag 
               
               
                   
               
               
                 5581 
                 agtcggtgaa gggcatgagg aagagcgagc ggcgcatcaa ggagctcacc taccagacag 
               
               
                   
               
               
                 5641 
                 aggaagacaa aaagaacctg ctgcggctac aggacctggt ggacaagctg caactgaagg 
               
               
                   
               
               
                 5701 
                 tcaaggccta caagcgccag gccgaggagg cggaggagca agccaacacc aacctgtcca 
               
               
                   
               
               
                 5761 
                 agttccgcaa ggtgcagcat gagctggatg aggcagagga gcgggcggac atcgctgagt 
               
               
                   
               
               
                 5821 
                 cccaggtcaa caagcttcga gccaagagcc gtgacattgg tgccaagcaa aaaatgcacg 
               
               
                   
               
               
                 5881 
                 atgaggagtg acactgcctc gggaacctca ctcttgccaa cctgtaataa atatgagtgc 
               
               
                   
               
               
                 5941 
                 c 
               
            
           
         
       
     
     By “MLC2A polypeptide” or “human MLSC2A (hMLC2A) polypeptide” is meant a protein or fragment thereof having at least 85% amino acid sequence identity to the sequence provided at NCBI Accession No. NP_067046.1 and having calcium binding activity. The amino acid sequence provided at NCBI Accession No. NP_067046.1 is shown below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 27) 
                   
               
            
           
           
               
               
               
            
               
                   1 
                 masrkagtrg kvaatkqaqr gssnvfsmfe qaqiqefkea fscidqnrdg iickadlret 
                   
               
               
                   
               
               
                  61 
                 ysqlgkvsvp eeeldamlqe gkgpinftvf ltlfgeklng tdpeeailsa frmfdpsgkg 
               
               
                   
               
               
                 121 
                 vvnkdefkql lltqadkfsp aeveqmfalt pmdlagnidy kslcyiithg dekee 
               
            
           
         
       
     
     By “MLC2A polynucleotide” is meant a polynucleotide encoding a MLC2A polypeptide or fragment thereof. An exemplary MLC2A polynucleotide sequence is provided at NCBI Ref: NM_021223.2. The sequence provided at NCBI Ref: NM_021223.2 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 28) 
                   
               
            
           
           
               
               
               
            
               
                   1 
                 tctgcagaga gaatggccag caggaaggcg gggacccggg gcaaggtggc agccaccaag 
                   
               
               
                   
               
               
                  61 
                 caggcccaac gtggttcttc caacgtcttt tccatgtttg aacaagccca gatacaggag 
               
               
                   
               
               
                 121 
                 ttcaaagaag ccttcagctg tatcgaccag aatcgtgatg gcatcatctg caaggcagac 
               
               
                   
               
               
                 181 
                 ctgagggaga cctactccca gctggggaag gtgagtgtcc cagaggagga gctggacgcc 
               
               
                   
               
               
                 241 
                 atgctgcaag agggcaaggg ccccatcaac ttcaccgtct tcctcacgct ctttggggag 
               
               
                   
               
               
                 301 
                 aagctcaatg ggacagaccc cgaggaagcc atcctgagtg ccttccgcat gtttgacccc 
               
               
                   
               
               
                 361 
                 agcggcaaag gggtggtgaa caaggatgag ttcaagcagc ttctcctgac ccaggcagac 
               
               
                   
               
               
                 421 
                 aagttctctc cagctgaggt ggagcagatg ttcgccctga cacccatgga cctggcgggg 
               
               
                   
               
               
                 481 
                 aacatcgact acaagtcact gtgctacatc atcacccatg gagacgagaa agaggaatga 
               
               
                   
               
               
                 541 
                 ggggcagggc caggcccacg ggggggcacc tcaataaact ctgttgcaaa attggaaaaa 
               
               
                   
               
               
                 601 
                 aaaaaaaaaa aaaaaaaaa 
               
            
           
         
       
     
     By “MUC2 polypeptide” is meant a protein or fragment thereof having at least 85% amino acid sequence identity to the sequence provided at NCBI Accession No. NP_002448.3 and having and having a biological activity of a MUC2 polypeptide. Exemplary biological activities of a MUC2 polypeptide include polymerization into a gel and coating of epithelia of the intestines and other mucus membrane-containing organs. The amino acid sequence provided at NCBI Accession No. NP_002448.3 is shown below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 29) 
                   
               
            
           
           
               
               
               
            
               
                   1 
                 mglplarlaa vclalslagg selqtegrtr nhghnvcstw gnfhyktfdg dvfrfpgpcd 
                   
               
               
                   
               
               
                  61 
                 ynfasdcrgs ykefavhlkr gpgqaeapag vesilltikd dtiyltrhla vlngavvstp 
               
               
                   
               
               
                  121 
                 hyspglliek sdaytkvysr agltlmwnre dalmleldtk frnhtcglcg dynglqsyse 
               
               
                   
               
               
                  181 
                 flsdgvlfsp lefgnmqkin qpdvvcedpe eevapascse hraecerllt aeafadcgdl 
               
               
                   
               
               
                  241 
                 vplepylrac qqdrcrcpgg dtcvcstvae fsrqcshagg rpgnwrtatl cpktcpgnlv 
               
               
                   
               
               
                  301 
                 ylesgspcmd tcshlevssl ceehrmdgcf cpegtvyddi gdsgcvpvsq chcrlhghly 
               
               
                   
               
               
                  361 
                 tpgqeitndc eqcvcnagrw vckdlpcpgt caleggshit tfdgktytfh gdcyyvlakg 
               
               
                   
               
               
                  421 
                 dhndsyallg elapcgstdk qtclktvvll adkkknvvvf ksdgsvllne lqvnlphvta 
               
               
                   
               
               
                  481 
                 sfsvfrpssy himvsmaigv rlqvglapvm qlfvtldgas qgqvgglogn fnglegddfk 
               
               
                   
               
               
                  541 
                 tasglveatg agfantwkaq stchdkldwl ddpcslnies anyaehwcsl lkktetpfgr 
               
               
                   
               
               
                  601 
                 chsavdpaey ykrckydtcn cqnnedclca alssyaract akgvmlwgwr ehvcnkdvgs 
               
               
                   
               
               
                  661 
                 cpnsqvflyn lttcqqtcrs lseadshcle gfapvdgcgc pdhtfldekg rcvplakcsc 
               
               
                   
               
               
                  721 
                 yhrglyleag dvvvrqeerc vcrdgrlhcr qirligqsct apkihmdcsn ltalatskpr 
               
               
                   
               
               
                  781 
                 alscqtlaag yyhtecvsgc vcpdglmddg rggcvvekec pcvhnndlys sgakikvdcn 
               
               
                   
               
               
                  841 
                 tctckrgrwv ctqavchgtc siygsghyit fdgkyydfdg hcsyvavgdy cgqnsslgsf 
               
               
                   
               
               
                  901 
                 siitenvpcg ttgvtcskai kifmgrtelk ledkhrvviq rdeghhvayt trevgqylvv 
               
               
                   
               
               
                  961 
                 esstgiiviw dkrttvfikl apsykgtvcg lcgnfdhrsn ndfttrdhmv vsseldfgns 
               
               
                   
               
               
                 1021 
                 wkeaptcpdv stnpepcsln phrrswaekq csilkssvfs ichskvdpkp fyeacvhdsc 
               
               
                   
               
               
                 1081 
                 scdtggdcec fcsavasyaq ectkegacvf wrtpdlcpif cdyynpphec ewhyepcgnr 
               
               
                   
               
               
                 1141 
                 sfetcrting ihsnisysyl egcyprcpkd rpiyeedlkk cvtadkcgcy vedthyppga 
               
               
                   
               
               
                 1201 
                 svpteetcks cvctnssqvv crpeegkiln qtqdgafcyw eicgpngtve khfnicsitt 
               
               
                   
               
               
                 1261 
                 rpstlttftt itlpttpttf ttttttttpt sstvlsttpk lcclwsdwin edhpssgsdd 
               
               
                   
               
               
                 1321 
                 gdretfdgvc gapediecrs vkdphlsleq lgqkvqcdvs vgficknedq fgngpfglcy 
               
               
                   
               
               
                 1381 
                 dykirvnccw pmdkcittps pptttpsppp tstttlpptt tpsppttttt tppptttpsp 
               
               
                   
               
               
                 1441 
                 pitttttppp tttpsppist tttppptttp spptttpspp tttpsppttt tttppptttp 
               
               
                   
               
               
                 1501 
                 spptttpitp pastttlppt ttpspptttt ttppptttps pptttpitpp tstttlpptt 
               
               
                   
               
               
                 1561 
                 tpsppptttt tppptttpsp pttttpsppt ittttppptt tpsppttttt tppptttpsp 
               
               
                   
               
               
                 1621 
                 ptttpitppt stttlppttt pspppttttt ppptttpspp ttttpsppit ttttpppttt 
               
               
                   
               
               
                 1681 
                 psspitttps pptttmttps ptttpsspit ttttpssttt pspppttmtt psptttpspp 
               
               
                   
               
               
                 1741 
                 tttmttlppt ttsspltttp lppsitpptf spfstttptt pcvplcnwtg wldsgkpnfh 
               
               
                   
               
               
                 1801 
                 kpggdtelig dvcgpgwaan iscratmypd vpigglgqtv vcdvsvglic knedqkpggv 
               
               
                   
               
               
                 1861 
                 ipmafclnye invqccecvt qpttmttttt enptpptttp itttttvtpt ptptgtqtpt 
               
               
                   
               
               
                 1921 
                 ttpitttttv tptptptgtq tptttpittt ttvtptptpt gtqtptttpi tttttvtptp 
               
               
                   
               
               
                 1981 
                 tptgtqtptt tpitttttvt ptptptgtqt ptttpitttt tvtptptptg tqtptttpit 
               
               
                   
               
               
                 2041 
                 ttttvtptpt ptgtqtpttt pitttttvtp tptptgtqtp tttpittttt vtptptptgt 
               
               
                   
               
               
                 2101 
                 qtptttpitt tttvtptptp tgtqtptttp itttttvtpt ptptgtqtpt ttpitttttv 
               
               
                   
               
               
                 2161 
                 tptptptgtq tptttpittt ttvtptptpt gtqtptttpi tttttvtptp tptgtqtptt 
               
               
                   
               
               
                 2221 
                 tpitttttvt ptptptgtqt ptttpitttt tvtptptptg tqtptttpit ttttvtptpt 
               
               
                   
               
               
                 2281 
                 ptgtqtpttt pitttttvtp tptptgtqtp tttpittttt vtptptptgt qtptttpitt 
               
               
                   
               
               
                 2341 
                 tttvtptptp tgtqtptttp itttttvtpt ptptgtqtpt ttpitttttv tptptptgtq 
               
               
                   
               
               
                 2401 
                 tptttpittt ttvtptptpt gtqtptttpi tttttvtptp tptgtqtptt tpitttttvt 
               
               
                   
               
               
                 2461 
                 ptptptgtqt ptttpitttt tvtptptptg tqtptttpit ttttvtptpt ptgtqtpttt 
               
               
                   
               
               
                 2521 
                 pitttttvtp tptptgtqtp tttpittttt vtptptptgt qtptttpitt tttvtptptp 
               
               
                   
               
               
                 2581 
                 tgtqtptttp itttttvtpt ptptgtqtpt ttpitttttv tptptptgtq tptttpittt 
               
               
                   
               
               
                 2641 
                 ttvtptptpt gtqtptttpi tttttvtptp tptgtqtptt tpitttttvt ptptptgtqt 
               
               
                   
               
               
                 2701 
                 ptttpitttt tvtptptptg tqtptttpit ttttvtptpt ptgtqtpttt pitttttvtp 
               
               
                   
               
               
                 2761 
                 tptptgtqtp tttpittttt vtptptptgt qtptttpitt tttvtptptp tgtqtptttp 
               
               
                   
               
               
                 2821 
                 itttttvtpt ptptgtqtpt ttpitttttv tptptptgtq tptttpittt ttvtptptpt 
               
               
                   
               
               
                 2881 
                 gtqtptttpi tttttvtptp tptgtqtptt tpitttttvt ptptptgtqt ptttpitttt 
               
               
                   
               
               
                 2941 
                 tvtptptptg tqtptttpit ttttvtptpt ptgtqtpttt pitttttvtp tptptgtqtp 
               
               
                   
               
               
                 3001 
                 tttpittttt vtptptptgt qtptttpitt tttvtptptp tgtqtptttp itttttvtpt 
               
               
                   
               
               
                 3061 
                 ptptgtqtpt ttpitttttv tptptptgtq tptttpittt ttvtptptpt gtqtptttpi 
               
               
                   
               
               
                 3121 
                 tttttvtptp tptgtqtptt tpitttttvt ptptptgtqt ptttpitttt tvtptptptg 
               
               
                   
               
               
                 3181 
                 tqtptttpit ttttvtptpt ptgtqtpttt pitttttvtp tptptgtqtp tttpittttt 
               
               
                   
               
               
                 3241 
                 vtptptptgt qtptttpitt tttvtptptp tgtqtptttp itttttvtpt ptptgtqtpt 
               
               
                   
               
               
                 3301 
                 ttpitttttv tptptptgtq tptttpittt ttvtptptpt gtqtptttpi tttttvtptp 
               
               
                   
               
               
                 3361 
                 tptgtqtptt tpitttttvt ptptptgtqt ptttpitttt tvtptptptg tqtptttpit 
               
               
                   
               
               
                 3421 
                 ttttvtptpt ptgtqtpttt pitttttvtp tptptgtqtp tttpittttt vtptptptgt 
               
               
                   
               
               
                 3481 
                 qtptttpitt tttvtptptp tgtqtptttp itttttvtpt ptptgtqtpt ttpitttttv 
               
               
                   
               
               
                 3541 
                 tptptptgtq tptttpittt ttvtptptpt gtqtptttpi tttttvtptp tptgtqtptt 
               
               
                   
               
               
                 3601 
                 tpitttttvt ptptptgtqt ptttpitttt tvtptptptg tqtptttpit ttttvtptpt 
               
               
                   
               
               
                 3661 
                 ptgtqtpttt pitttttvtp tptptgtqtp tttpittttt vtptptptgt qtptttpitt 
               
               
                   
               
               
                 3721 
                 tttvtptptp tgtqtptttp itttttvtpt ptptgtqtpt ttpitttttv tptptptgtq 
               
               
                   
               
               
                 3781 
                 tptttpittt ttvtptptpt gtqtptttpi tttttvtptp tptgtqtptt tpitttttvt 
               
               
                   
               
               
                 3841 
                 ptptptgtqt ptttpitttt tvtptptptg tqtptttpit ttttvtptpt ptgtqtpttt 
               
               
                   
               
               
                 3901 
                 pitttttvtp tptptgtqtp tttpittttt vtptptptgt qtptttpitt tttvtptptp 
               
               
                   
               
               
                 3961 
                 tgtqtptttp itttttvtpt ptptgtqtpt ttpitttttv tptptptgtq tptttpittt 
               
               
                   
               
               
                 4021 
                 ttvtptptpt gtqtptttpi tttttvtptp tptgtqtptt tpitttttvt ptptptgtqt 
               
               
                   
               
               
                 4081 
                 ptttpitttt tvtptptptg tqtptttpit ttttvtptpt ptgtqtpttt pitttttvtp 
               
               
                   
               
               
                 4141 
                 tptptgtqtp tttpittttt vtptptptgt qtptttpitt tttvtptptp tgtqtgppth 
               
               
                   
               
               
                 4201 
                 tstapiaelt tsnpppesst pqtsrstssp ltesttllst lppaiemtst appstptapt 
               
               
                   
               
               
                 4261 
                 ttsgghtlsp ppstttsppg tptrgtttgs ssaptpstvq ttttsawtpt ptplstpsii 
               
               
                   
               
               
                 4321 
                 rttglrpyps svliccvind tyyapgeevy ngtygdtcyf vncslsctle fynwscpstp 
               
               
                   
               
               
                 4381 
                 sptptpskst ptpskpsstp skptpgtkpp ecpdfdpprq enetwwlcdc fmatckynnt 
               
               
                   
               
               
                 4441 
                 veivkvecep ppmptcsngl qpvrvedpdg ccwhwecdcy ctgwgdphyv tfdglyysyq 
               
               
                   
               
               
                 4501 
                 gnctyvlvee ispsvdnfgv yidnyhcdpn dkvscprtli vrhetqevli ktvhmmpmqv 
               
               
                   
               
               
                 4561 
                 qvgvnrgava lpykkyglev yqsginyvvd ipelgvlvsy nglsfsvrlp yhrfgnntkg 
               
               
                   
               
               
                 4621 
                 qcgtctntts ddcilpsgei vsnceaaadq wlvndpskph cphsssttkr pavtvpgggk 
               
               
                   
               
               
                 4681 
                 ttphkdctps ploglikdsl faqchalvpp qhyydacvfd scfmpgssle caslqayaal 
               
               
                   
               
               
                 4741 
                 caqqnicldw rnhthgaclv ecpshreyqa cgpaeeptck ssssqqnntv lvegcfcpeg 
               
               
                   
               
               
                 4801 
                 tmnyapgfdv cvktcgcvgp dnvprefgeh fefdckncvc leggsgiicq pkrcsqkpvt 
               
               
                   
               
               
                 4861 
                 hcvedgtyla tevnpadtcc nitvckcnts lckekpsvcp lgfevkskmv pgrccpfywc 
               
               
                   
               
               
                 4921 
                 eskgvcvhgn aeyqpgspvy sskcqdcvct dkvdnntlln viacthvpcn tscspgfelm 
               
               
                   
               
               
                 4981 
                 eapgecckkc eqthciikrp dnqhvilkpg dfksdpknnc tffscvkihn qlissysnit 
               
               
                   
               
               
                 5041 
                 cpnfdasici pgsitfmpng ccktctprne trvpcstvpv ttevsyagct ktvlmnhcsg 
               
               
                   
               
               
                 5101 
                 scgtfvmysa kaqaldhscs cckeektsqr evvlscpngg slthtythie scqcqdtvcg 
               
               
                   
               
               
                 5161 
                 lptgtsrrar rsprhlgsg 
               
            
           
         
       
     
     By “MUC2 polynucleotide” is meant a polynucleotide encoding a MUC2 polypeptide or fragment thereof. An exemplary MUC2 polynucleotide sequence is provided at NCBI Ref: NM_002457.3. The sequence provided at NCBI Ref: NM_002457.3 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 30) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 caacccacac cgcccctgcc agccaccatg gggctgccac tagcccgcct ggcggctgtg 
                   
               
               
                   
               
               
                 61 
                 tgcctggccc tgtctttggc agggggctcg gagctccaga cagagggcag aacccgaaac 
               
               
                   
               
               
                 121 
                 cacggccaca acgtctgcag cacctggggc aacttccact acaagacctt cgacggggac 
               
               
                   
               
               
                 181 
                 gtcttccgct tccccggccc ctgcgactac aacttcgcct ccgactgccg aggctcctac 
               
               
                   
               
               
                 241 
                 aaggaatttg ctgtgcacct gaagcggggt ccgggccagg ctgaggcccc cgccggggtg 
               
               
                   
               
               
                 301 
                 gagtccatcc tgctgaccat caaggatgac accatctacc tcacccgcca cctggctgtg 
               
               
                   
               
               
                 361 
                 cttaacgggg ccgtggtcag caccccgcac tacagccccg ggctgctcat tgagaagagc 
               
               
                   
               
               
                 421 
                 gatgcctaca ccaaagtcta ctcccgcgcc ggcctcaccc tcatgtggaa ccgggaggat 
               
               
                   
               
               
                 481 
                 gcactcatgc tggagctgga cactaagttc cggaaccaca cctgtggcct ctgcggggac 
               
               
                   
               
               
                 541 
                 tacaacggcc tgcagagcta ttcagaattc ctctctgacg gcgtgctctt cagtcccctg 
               
               
                   
               
               
                 601 
                 gagtttggga acatgcagaa gatcaaccag cccgatgtgg tgtgtgagga tcccgaggag 
               
               
                   
               
               
                 661 
                 gaggtggccc ccgcatcctg ctccgagcac cgcgccgagt gtgagaggct gctgaccgcc 
               
               
                   
               
               
                 721 
                 gaggccttcg cggactgtca ggacctggtg ccgctggagc cgtatctgcg cgcctgccag 
               
               
                   
               
               
                 781 
                 caggaccgct gccggtgccc gggcggtgac acctgcgtct gcagcaccgt ggccgagttc 
               
               
                   
               
               
                 841 
                 tcccgccagt gctcccacgc cggcggccgg cccgggaact ggaggaccgc cacgctctgc 
               
               
                   
               
               
                 901 
                 cccaagacct gccccgggaa cctggtgtac ctggagagcg gctcgccctg catggacacc 
               
               
                   
               
               
                 961 
                 tgctcacacc tggaggtgag cagcctgtgc gaggagcacc gcatggacgg ctgtttctgc 
               
               
                   
               
               
                 1021 
                 ccagaaggca ccgtatatga cgacatcggg gacagtggct gcgttcctgt gagccagtgc 
               
               
                   
               
               
                 1081 
                 cactgcaggc tgcacggaca cctgtacaca ccgggccagg agatcaccaa tgactgcgag 
               
               
                   
               
               
                 1141 
                 cagtgtgtct gtaacgctgg ccgctgggtg tgcaaagacc tgccctgccc cggcacctgt 
               
               
                   
               
               
                 1201 
                 gccctggaag gcggctccca catcaccacc ttcgatggga agacgtacac cttccacggg 
               
               
                   
               
               
                 1261 
                 gactgctact atgtcctggc caagggtgac cacaacgatt cctacgctct cctgggcgag 
               
               
                   
               
               
                 1321 
                 ctggccccct gtggctccac agacaagcag acctgcctga agacggtggt gctgctggct 
               
               
                   
               
               
                 1381 
                 gacaagaaga agaatgtggt ggtcttcaag tccgatggca gtgtactgct caacgagctg 
               
               
                   
               
               
                 1441 
                 caggtgaacc tgccccacgt gaccgcgagc ttctctgtct tccgcccgtc ttcctaccac 
               
               
                   
               
               
                 1501 
                 atcatggtga gcatggccat tggcgtccgg ctgcaggtgc agctggcccc agtcatgcaa 
               
               
                   
               
               
                 1561 
                 ctctttgtga cactggacca ggcctcccag gggcaggtgc agggcctctg cgggaacttc 
               
               
                   
               
               
                 1621 
                 aacggcctgg aaggtgacga cttcaagacg gccagcgggc tggtggaggc cacgggggcc 
               
               
                   
               
               
                 1681 
                 ggctttgcca acacctggaa ggcacagtca acctgccatg acaagctgga ctggttggac 
               
               
                   
               
               
                 1741 
                 gatccctgct ccctgaacat cgagagcgcc aactacgccg agcactggtg ctccctcctg 
               
               
                   
               
               
                 1801 
                 aagaagacag agaccccctt tggcaggtgc cactcggctg tggaccctgc tgagtattac 
               
               
                   
               
               
                 1861 
                 aagaggtgca aatatgacac gtgtaactgt cagaacaatg aggactgcct gtgcgccgcc 
               
               
                   
               
               
                 1921 
                 ctgtcctcct acgcgcgcgc ctgcaccgcc aagggcgtca tgctgtgggg ctggcgggag 
               
               
                   
               
               
                 1981 
                 catgtctgca acaaggatgt gggctcctgc cccaactcgc aggtcttcct gtacaacctg 
               
               
                   
               
               
                 2041 
                 accacctgcc agcagacctg ccgctccctc tccgaggccg acagccactg tctcgagggc 
               
               
                   
               
               
                 2101 
                 tttgcgcctg tggacggctg cggctgccct gaccacacct tcctggacga gaagggccgc 
               
               
                   
               
               
                 2161 
                 tgcgtacccc tggccaagtg ctcctgttac caccgcggtc tctacctgga ggcgggggac 
               
               
                   
               
               
                 2221 
                 gtggtcgtca ggcaggaaga acgatgtgtg tgccgggatg ggcggctgca ctgtaggcag 
               
               
                   
               
               
                 2281 
                 atccggctga tcggccagag ctgcacggcc ccaaagatcc acatggactg cagcaacctg 
               
               
                   
               
               
                 2341 
                 actgcactgg ccacctcgaa gccccgagcc ctcagctgcc agacgctggc cgccggctat 
               
               
                   
               
               
                 2401 
                 taccacacag agtgtgtcag tggctgtgtg tgccccgacg ggctgatgga tgacggccgg 
               
               
                   
               
               
                 2461 
                 ggtggctgcg tggtggagaa ggaatgccct tgcgtccata acaacgacct gtattcttcc 
               
               
                   
               
               
                 2521 
                 ggcgccaaga tcaaggtgga ctgcaatacc tgcacctgca agagaggacg ctgggtgtgc 
               
               
                   
               
               
                 2581 
                 acccaggctg tgtgccatgg cacctgctcc atttacggga gtggccacta catcaccttt 
               
               
                   
               
               
                 2641 
                 gacgggaagt actacgactt tgacggacac tgctcctacg tggctgttca ggactactgc 
               
               
                   
               
               
                 2701 
                 ggccagaact cctcactggg ctcattcagc atcatcaccg agaacgtccc ctgtggcact 
               
               
                   
               
               
                 2761 
                 acgggcgtca cctgctccaa ggccatcaag atcttcatgg ggaggacgga gctgaagttg 
               
               
                   
               
               
                 2821 
                 gaagacaagc accgtgtggt gatccagcgt gatgagggtc accacgtggc ctacaccacg 
               
               
                   
               
               
                 2881 
                 cgggaggtgg gccagtacct ggtggtggag tccagcacgg gcatcatcgt catctgggac 
               
               
                   
               
               
                 2941 
                 aagaggacca ccgtgttcat caagctggct ccctcctaca agggcaccgt gtgtggcctg 
               
               
                   
               
               
                 3001 
                 tgtgggaact ttgaccaccg ctccaacaac gacttcacca cgcgggacca catggtggtg 
               
               
                   
               
               
                 3061 
                 agcagcgagc tggacttcgg gaacagctgg aaggaggccc ccacctgccc agatgtgagc 
               
               
                   
               
               
                 3121 
                 accaaccccg agccctgcag cctgaacccg caccgccgct cctgggccga gaagcagtgc 
               
               
                   
               
               
                 3181 
                 agcatcctca aaagcagcgt gttcagcatc tgccacagca aggtggaccc caagcccttc 
               
               
                   
               
               
                 3241 
                 tacgaggcct gtgtgcacga ctcgtgctcc tgtgacacgg gtggggactg tgagtgcttc 
               
               
                   
               
               
                 3301 
                 tgctctgccg tggcctccta cgcccaggag tgtaccaaag agggggcctg cgtgttctgg 
               
               
                   
               
               
                 3361 
                 aggacgccgg acctgtgccc catattctgc gactactaca accctccgca tgagtgtgag 
               
               
                   
               
               
                 3421 
                 tggcactatg agccatgtgg gaaccggagc ttcgagacct gcaggaccat caatggcatc 
               
               
                   
               
               
                 3481 
                 cactccaaca tctccgtgtc ctacctggag ggctgctacc cccggtgccc caaggacagg 
               
               
                   
               
               
                 3541 
                 cccatctatg aggaggatct gaagaagtgt gtcactgcag acaagtgtgg ctgctatgtc 
               
               
                   
               
               
                 3601 
                 gaggacaccc actacccacc tggagcatcg gttcccaccg aggagacctg caagtcctgc 
               
               
                   
               
               
                 3661 
                 gtgtgtacca actcctccca agtcgtctgc aggccggagg aaggaaagat tcttaaccag 
               
               
                   
               
               
                 3721 
                 acccaggatg gcgccttctg ctactgggag atctgtggcc ccaacgggac ggtggagaag 
               
               
                   
               
               
                 3781 
                 cacttcaaca tctgttccat tacgacacgc ccgtccaccc tgaccacctt caccaccatc 
               
               
                   
               
               
                 3841 
                 accctcccca ccacccccac caccttcacc actaccacca ccaccaccac cccgacctcc 
               
               
                   
               
               
                 3901 
                 agcacagttt tatcaacaac tccgaagctg tgctgcctct ggtctgactg gatcaatgag 
               
               
                   
               
               
                 3961 
                 gaccacccca gcagtggcag cgacgacggt gaccgagaaa catttgatgg ggtctgcggg 
               
               
                   
               
               
                 4021 
                 gcccctgagg acatcgagtg caggtcggtc aaggatcccc acctcagctt ggagcagcta 
               
               
                   
               
               
                 4081 
                 ggccagaagg tgcagtgtga tgtctctgtt gggttcattt gcaagaatga agaccagttt 
               
               
                   
               
               
                 4141 
                 ggaaatggac catttggact gtgttacgac tacaagatac gtgtcaattg ttgctggccc 
               
               
                   
               
               
                 4201 
                 atggataagt gtatcaccac tcccagccct ccaactacca ctcccagccc tccaccaacc 
               
               
                   
               
               
                 4261 
                 agcacgacca cccttccacc aaccaccacc cccagccctc caaccaccac cacaaccacc 
               
               
                   
               
               
                 4321 
                 cctccaccaa ccaccacccc cagccctcca ataaccacca cgaccacccc tccaccaacc 
               
               
                   
               
               
                 4381 
                 accactccca gccctccaat aagcaccaca accacccctc caccaaccac cactcccagc 
               
               
                   
               
               
                 4441 
                 cctccaacca ccactcccag ccctccaacc accactccca gccctccaac aaccaccaca 
               
               
                   
               
               
                 4501 
                 accacccctc caccaaccac cactcccagc cctccaacga ctacgcccat cactccacca 
               
               
                   
               
               
                 4561 
                 gccagcacta ccacccttcc accaaccacc actcccagcc ctccaacaac caccacaacc 
               
               
                   
               
               
                 4621 
                 acccctccac caaccaccac tcccagtcct ccaacgacta cgcccatcac tccaccaacc 
               
               
                   
               
               
                 4681 
                 agcactacta cccttccacc aaccaccact cccagccctc caccaaccac cacaaccacc 
               
               
                   
               
               
                 4741 
                 cctccaccaa ccaccactcc cagccctcca acaaccacca ctcccagtcc tccaacaatc 
               
               
                   
               
               
                 4801 
                 accacaacca cccctccacc aaccaccact cccagccctc caacaacgac cacaaccacc 
               
               
                   
               
               
                 4861 
                 cctccaccaa ccaccactcc cagccctcca acgactacac ccatcactcc accaaccagc 
               
               
                   
               
               
                 4921 
                 actaccaccc ttccaccaac caccactccc agccctccac caaccaccac aaccacccct 
               
               
                   
               
               
                 4981 
                 ccaccaacca ccactcccag ccctccaaca accaccactc ccagccctcc aataaccacc 
               
               
                   
               
               
                 5041 
                 acaaccaccc ctccaccaac caccactccc agctctccaa taaccaccac tcccagccct 
               
               
                   
               
               
                 5101 
                 ccaacaacca ccatgaccac cccttcacca accaccaccc ccagctctcc aataaccacc 
               
               
                   
               
               
                 5161 
                 acaaccaccc cttcctcaac taccactccc agccctccac caaccaccat gaccacccct 
               
               
                   
               
               
                 5221 
                 tcaccaacca ccactcccag ccctccaaca accaccatga ccacccttcc accaaccacc 
               
               
                   
               
               
                 5281 
                 acttccagcc ctctaacaac tactcctcta cctccatcaa taactcctcc tacattttca 
               
               
                   
               
               
                 5341 
                 ccattctcaa cgacaacccc tactacccca tgcgtgcctc tctgcaattg gactggctgg 
               
               
                   
               
               
                 5401 
                 ctggattctg gaaaacccaa ctttcacaaa ccaggtggag acacagaatt gattggagac 
               
               
                   
               
               
                 5461 
                 gtctgtggac caggctgggc agctaacatc tcttgcagag ccaccatgta tcctgatgtt 
               
               
                   
               
               
                 5521 
                 cccattggac agcttggaca aacagtggtg tgtgatgtct ctgtggggct gatatgcaaa 
               
               
                   
               
               
                 5581 
                 aatgaagacc aaaagccagg tggggtcatc cctatggcct tctgcctcaa ctacgagatc 
               
               
                   
               
               
                 5641 
                 aacgttcagt gctgtgagtg tgtcacccaa cccaccacca tgacaaccac caccacagag 
               
               
                   
               
               
                 5701 
                 aacccaactc cgccaaccac gacacccatc accaccacca ctacggtgac cccaacccca 
               
               
                   
               
               
                 5761 
                 acacccaccg gcacacagac cccaaccacg acacccatca ccaccaccac tacggtgacc 
               
               
                   
               
               
                 5821 
                 ccaaccccaa cacccaccgg cacacagacc ccaaccacga cacccatcac caccaccact 
               
               
                   
               
               
                 5881 
                 acggtgaccc caaccccaac acccaccggc acacagaccc caaccacgac acccatcacc 
               
               
                   
               
               
                 5941 
                 accaccacta cggtgacccc aaccccaaca cccaccggca cacagacccc aaccacgaca 
               
               
                   
               
               
                 6001 
                 cccatcacca ccaccactac ggtgacccca accccaacac ccaccggcac acagacccca 
               
               
                   
               
               
                 6061 
                 accacgacac ccatcaccac caccactacg gtgaccccaa ccccaacacc caccggcaca 
               
               
                   
               
               
                 6121 
                 cagaccccaa ccacgacacc catcaccacc accactacgg tgaccccaac cccaacaccc 
               
               
                   
               
               
                 6181 
                 accggcacac agaccccaac cacgacaccc atcaccacca ccactacggt gaccccaacc 
               
               
                   
               
               
                 6241 
                 ccaacaccca ccggcacaca gaccccaacc acgacaccca tcaccaccac cactacggtg 
               
               
                   
               
               
                 6301 
                 accccaaccc caacacccac cggcacacag accccaacca cgacacccat caccaccacc 
               
               
                   
               
               
                 6361 
                 actacggtga ccccaacccc aacacccacc ggcacacaga ccccaaccac gacacccatc 
               
               
                   
               
               
                 6421 
                 accaccacca ctacggtgac cccaacccca acacccaccg gcacacagac cccaaccacg 
               
               
                   
               
               
                 6481 
                 acacccatca ccaccaccac tacggtgacc ccaaccccaa cacccaccgg cacacagacc 
               
               
                   
               
               
                 6541 
                 ccaaccacga cacccatcac caccaccact acggtgaccc caaccccaac acccaccggc 
               
               
                   
               
               
                 6601 
                 acacagaccc caaccacgac acccatcacc accaccacta cggtgacccc aaccccaaca 
               
               
                   
               
               
                 6661 
                 cccaccggca cacagacccc aaccacgaca cccatcacca ccaccactac ggtgacccca 
               
               
                   
               
               
                 6721 
                 accccaacac ccaccggcac acagacccca accacgacac ccatcaccac caccactacg 
               
               
                   
               
               
                 6781 
                 gtgaccccaa ccccaacacc caccggcaca cagaccccaa ccacgacacc catcaccacc 
               
               
                   
               
               
                 6841 
                 accactacgg tgaccccaac cccaacaccc accggcacac agaccccaac cacgacaccc 
               
               
                   
               
               
                 6901 
                 atcaccacca ccactacggt gaccccaacc ccaacaccca ccggcacaca gaccccaacc 
               
               
                   
               
               
                 6961 
                 acgacaccca tcaccaccac cactacggtg accccaaccc caacacccac cggcacacag 
               
               
                   
               
               
                 7021 
                 accccaacca cgacacccat caccaccacc actacggtga ccccaacccc aacacccacc 
               
               
                   
               
               
                 7081 
                 ggcacacaga ccccaaccac gacacccatc accaccacca ctacggtgac cccaacccca 
               
               
                   
               
               
                 7141 
                 acacccaccg gcacacagac cccaaccacg acacccatca ccaccaccac tacggtgacc 
               
               
                   
               
               
                 7201 
                 ccaaccccaa cacccaccgg cacacagacc ccaaccacga cacccatcac caccaccact 
               
               
                   
               
               
                 7261 
                 acggtgaccc caaccccaac acccaccggc acacagaccc caaccacgac acccatcacc 
               
               
                   
               
               
                 7321 
                 accaccacta cggtgacccc aaccccaaca cccaccggca cacagacccc aaccacgaca 
               
               
                   
               
               
                 7381 
                 cccatcacca ccaccactac ggtgacccca accccaacac ccaccggcac acagacccca 
               
               
                   
               
               
                 7441 
                 accacgacac ccatcaccac caccactacg gtgaccccaa ccccaacacc caccggcaca 
               
               
                   
               
               
                 7501 
                 cagaccccaa ccacgacacc catcaccacc accactacgg tgaccccaac cccaacaccc 
               
               
                   
               
               
                 7561 
                 accggcacac agaccccaac cacgacaccc atcaccacca ccactacggt gaccccaacc 
               
               
                   
               
               
                 7621 
                 ccaacaccca ccggcacaca gaccccaacc acgacaccca tcaccaccac cactacggtg 
               
               
                   
               
               
                 7681 
                 accccaaccc caacacccac cggcacacag accccaacca cgacacccat caccaccacc 
               
               
                   
               
               
                 7741 
                 actacggtga ccccaacccc aacacccacc ggcacacaga ccccaaccac gacacccatc 
               
               
                   
               
               
                 7801 
                 accaccacca ctacggtgac cccaacccca acacccaccg gcacacagac cccaaccacg 
               
               
                   
               
               
                 7861 
                 acacccatca ccaccaccac tacggtgacc ccaaccccaa cacccaccgg cacacagacc 
               
               
                   
               
               
                 7921 
                 ccaaccacga cacccatcac caccaccact acggtgaccc caaccccaac acccaccggc 
               
               
                   
               
               
                 7981 
                 acacagaccc caaccacgac acccatcacc accaccacta cggtgacccc aaccccaaca 
               
               
                   
               
               
                 8041 
                 cccaccggca cacagacccc aaccacgaca cccatcacca ccaccactac ggtgacccca 
               
               
                   
               
               
                 8101 
                 accccaacac ccaccggcac acagacccca accacgacac ccatcaccac caccactacg 
               
               
                   
               
               
                 8161 
                 gtgaccccaa ccccaacacc caccggcaca cagaccccaa ccacgacacc catcaccacc 
               
               
                   
               
               
                 8221 
                 accactacgg tgaccccaac cccaacaccc accggcacac agaccccaac cacgacaccc 
               
               
                   
               
               
                 8281 
                 atcaccacca ccactacggt gaccccaacc ccaacaccca ccggcacaca gaccccaacc 
               
               
                   
               
               
                 8341 
                 acgacaccca tcaccaccac cactacggtg accccaaccc caacacccac cggcacacag 
               
               
                   
               
               
                 8401 
                 accccaacca cgacacccat caccaccacc actacggtga ccccaacccc aacacccacc 
               
               
                   
               
               
                 8461 
                 ggcacacaga ccccaaccac gacacccatc accaccacca ctacggtgac cccaacccca 
               
               
                   
               
               
                 8521 
                 acacccaccg gcacacagac cccaaccacg acacccatca ccaccaccac tacggtgacc 
               
               
                   
               
               
                 8581 
                 ccaaccccaa cacccaccgg cacacagacc ccaaccacga cacccatcac caccaccact 
               
               
                   
               
               
                 8641 
                 acggtgaccc caaccccaac acccaccggc acacagaccc caaccacgac acccatcacc 
               
               
                   
               
               
                 8701 
                 accaccacta cggtgacccc aaccccaaca cccaccggca cacagacccc aaccacgaca 
               
               
                   
               
               
                 8761 
                 cccatcacca ccaccactac ggtgacccca accccaacac ccaccggcac acagacccca 
               
               
                   
               
               
                 8821 
                 accacgacac ccatcaccac caccactacg gtgaccccaa ccccaacacc caccggcaca 
               
               
                   
               
               
                 8881 
                 cagaccccaa ccacgacacc catcaccacc accactacgg tgaccccaac cccaacaccc 
               
               
                   
               
               
                 8941 
                 accggcacac agaccccaac cacgacaccc atcaccacca ccactacggt gaccccaacc 
               
               
                   
               
               
                 9001 
                 ccaacaccca ccggcacaca gaccccaacc acgacaccca tcaccaccac cactacggtg 
               
               
                   
               
               
                 9061 
                 accccaaccc caacacccac cggcacacag accccaacca cgacacccat caccaccacc 
               
               
                   
               
               
                 9121 
                 actacggtga ccccaacccc aacacccacc ggcacacaga ccccaaccac gacacccatc 
               
               
                   
               
               
                 9181 
                 accaccacca ctacggtgac cccaacccca acacccaccg gcacacagac cccaaccacg 
               
               
                   
               
               
                 9241 
                 acacccatca ccaccaccac tacggtgacc ccaaccccaa cacccaccgg cacacagacc 
               
               
                   
               
               
                 9301 
                 ccaaccacga cacccatcac caccaccact acggtgaccc caaccccaac acccaccggc 
               
               
                   
               
               
                 9361 
                 acacagaccc caaccacgac acccatcacc accaccacta cggtgacccc aaccccaaca 
               
               
                   
               
               
                 9421 
                 cccaccggca cacagacccc aaccacgaca cccatcacca ccaccactac ggtgacccca 
               
               
                   
               
               
                 9481 
                 accccaacac ccaccggcac acagacccca accacgacac ccatcaccac caccactacg 
               
               
                   
               
               
                 9541 
                 gtgaccccaa ccccaacacc caccggcaca cagaccccaa ccacgacacc catcaccacc 
               
               
                   
               
               
                 9601 
                 accactacgg tgaccccaac cccaacaccc accggcacac agaccccaac cacgacaccc 
               
               
                   
               
               
                 9661 
                 atcaccacca ccactacggt gaccccaacc ccaacaccca ccggcacaca gaccccaacc 
               
               
                   
               
               
                 9721 
                 acgacaccca tcaccaccac cactacggtg accccaaccc caacacccac cggcacacag 
               
               
                   
               
               
                 9781 
                 accccaacca cgacacccat caccaccacc actacggtga ccccaacccc aacacccacc 
               
               
                   
               
               
                 9841 
                 ggcacacaga ccccaaccac gacacccatc accaccacca ctacggtgac cccaacccca 
               
               
                   
               
               
                 9901 
                 acacccaccg gcacacagac cccaaccacg acacccatca ccaccaccac tacggtgacc 
               
               
                   
               
               
                 9961 
                 ccaaccccaa cacccaccgg cacacagacc ccaaccacga cacccatcac caccaccact 
               
               
                   
               
               
                 10021 
                 acggtgaccc caaccccaac acccaccggc acacagaccc caaccacgac acccatcacc 
               
               
                   
               
               
                 10081 
                 accaccacta cggtgacccc aaccccaaca cccaccggca cacagacccc aaccacgaca 
               
               
                   
               
               
                 10141 
                 cccatcacca ccaccactac ggtgacccca accccaacac ccaccggcac acagacccca 
               
               
                   
               
               
                 10201 
                 accacgacac ccatcaccac caccactacg gtgaccccaa ccccaacacc caccggcaca 
               
               
                   
               
               
                 10261 
                 cagaccccaa ccacgacacc catcaccacc accactacgg tgaccccaac cccaacaccc 
               
               
                   
               
               
                 10321 
                 accggcacac agaccccaac cacgacaccc atcaccacca ccactacggt gaccccaacc 
               
               
                   
               
               
                 10381 
                 ccaacaccca ccggcacaca gaccccaacc acgacaccca tcaccaccac cactacggtg 
               
               
                   
               
               
                 10441 
                 accccaaccc caacacccac cggcacacag accccaacca cgacacccat caccaccacc 
               
               
                   
               
               
                 10501 
                 actacggtga ccccaacccc aacacccacc ggcacacaga ccccaaccac gacacccatc 
               
               
                   
               
               
                 10561 
                 accaccacca ctacggtgac cccaacccca acacccaccg gcacacagac cccaaccacg 
               
               
                   
               
               
                 10621 
                 acacccatca ccaccaccac tacggtgacc ccaaccccaa cacccaccgg cacacagacc 
               
               
                   
               
               
                 10681 
                 ccaaccacga cacccatcac caccaccact acggtgaccc caaccccaac acccaccggc 
               
               
                   
               
               
                 10741 
                 acacagaccc caaccacgac acccatcacc accaccacta cggtgacccc aaccccaaca 
               
               
                   
               
               
                 10801 
                 cccaccggca cacagacccc aaccacgaca cccatcacca ccaccactac ggtgacccca 
               
               
                   
               
               
                 10861 
                 accccaacac ccaccggcac acagacccca accacgacac ccatcaccac caccactacg 
               
               
                   
               
               
                 10921 
                 gtgaccccaa ccccaacacc caccggcaca cagaccccaa ccacgacacc catcaccacc 
               
               
                   
               
               
                 10981 
                 accactacgg tgaccccaac cccaacaccc accggcacac agaccccaac cacgacaccc 
               
               
                   
               
               
                 11041 
                 atcaccacca ccactacggt gaccccaacc ccaacaccca ccggcacaca gaccccaacc 
               
               
                   
               
               
                 11101 
                 acgacaccca tcaccaccac cactacggtg accccaaccc caacacccac cggcacacag 
               
               
                   
               
               
                 11161 
                 accccaacca cgacacccat caccaccacc actacggtga ccccaacccc aacacccacc 
               
               
                   
               
               
                 11221 
                 ggcacacaga ccccaaccac gacacccatc accaccacca ctacggtgac cccaacccca 
               
               
                   
               
               
                 11281 
                 acacccaccg gcacacagac cccaaccacg acacccatca ccaccaccac tacggtgacc 
               
               
                   
               
               
                 11341 
                 ccaaccccaa cacccaccgg cacacagacc ccaaccacga cacccatcac caccaccact 
               
               
                   
               
               
                 11401 
                 acggtgaccc caaccccaac acccaccggc acacagaccc caaccacgac acccatcacc 
               
               
                   
               
               
                 11461 
                 accaccacta cggtgacccc aaccccaaca cccaccggca cacagacccc aaccacgaca 
               
               
                   
               
               
                 11521 
                 cccatcacca ccaccactac ggtgacccca accccaacac ccaccggcac acagacccca 
               
               
                   
               
               
                 11581 
                 accacgacac ccatcaccac caccactacg gtgaccccaa ccccaacacc caccggcaca 
               
               
                   
               
               
                 11641 
                 cagaccccaa ccacgacacc catcaccacc accactacgg tgaccccaac cccaacaccc 
               
               
                   
               
               
                 11701 
                 accggcacac agaccccaac cacgacaccc atcaccacca ccactacggt gaccccaacc 
               
               
                   
               
               
                 11761 
                 ccaacaccca ccggcacaca gaccccaacc acgacaccca tcaccaccac cactacggtg 
               
               
                   
               
               
                 11821 
                 accccaaccc caacacccac cggcacacag accccaacca cgacacccat caccaccacc 
               
               
                   
               
               
                 11881 
                 actacggtga ccccaacccc aacacccacc ggcacacaga ccccaaccac gacacccatc 
               
               
                   
               
               
                 11941 
                 accaccacca ctacggtgac cccaacccca acacccaccg gcacacagac cccaaccacg 
               
               
                   
               
               
                 12001 
                 acacccatca ccaccaccac tacggtgacc ccaaccccaa cacccaccgg cacacagacc 
               
               
                   
               
               
                 12061 
                 ccaaccacga cacccatcac caccaccact acggtgaccc caaccccaac acccaccggc 
               
               
                   
               
               
                 12121 
                 acacagaccc caaccacgac acccatcacc accaccacta cggtgacccc aaccccaaca 
               
               
                   
               
               
                 12181 
                 cccaccggca cacagacccc aaccacgaca cccatcacca ccaccactac ggtgacccca 
               
               
                   
               
               
                 12241 
                 accccaacac ccaccggcac acagacccca accacgacac ccatcaccac caccactacg 
               
               
                   
               
               
                 12301 
                 gtgaccccaa ccccaacacc caccggcaca cagaccccaa ccacgacacc catcaccacc 
               
               
                   
               
               
                 12361 
                 accactacgg tgaccccaac cccaacaccc accggcacac agaccccaac cacgacaccc 
               
               
                   
               
               
                 12421 
                 atcaccacca ccactacggt gaccccaacc ccaacaccca ccggcacaca gaccccaacc 
               
               
                   
               
               
                 12481 
                 acgacaccca tcaccaccac cactacggtg accccaaccc caacacccac cggcacacag 
               
               
                   
               
               
                 12541 
                 accccaacca cgacacccat caccaccacc actacggtga ccccaacccc aacacccacc 
               
               
                   
               
               
                 12601 
                 ggcacacaga ccgggccccc cacccacaca agcacagcac cgattgctga gttgaccaca 
               
               
                   
               
               
                 12661 
                 tccaatcctc cgcctgagtc ctcaacccct cagacctctc ggtccacctc ttcccctctc 
               
               
                   
               
               
                 12721 
                 acggagtcaa ccacccttct gagtacccta ccacctgcca ttgagatgac cagcacggcc 
               
               
                   
               
               
                 12781 
                 ccaccctcca cacccacggc acccacgacc acgagcggag gccacacact gtctccaccg 
               
               
                   
               
               
                 12841 
                 cccagcacca ccacgtcccc tccaggcacc cccactcgcg gtaccacgac tgggtcatct 
               
               
                   
               
               
                 12901 
                 tcagccccca cccccagcac tgtgcagacg accaccacca gtgcctggac ccccacgccg 
               
               
                   
               
               
                 12961 
                 accccactct ccacacccag catcatcagg accacaggcc tgaggcccta cccttcctct 
               
               
                   
               
               
                 13021 
                 gtgcttatct gctgtgtcct gaacgacacc tactacgcac caggtgagga ggtgtacaac 
               
               
                   
               
               
                 13081 
                 ggcacatacg gagacacctg ttatttcgtc aactgctcac tgagctgtac gttggagttc 
               
               
                   
               
               
                 13141 
                 tataactggt cctgcccatc cacgccctcc ccaacaccca cgccctccaa gtcgacgccc 
               
               
                   
               
               
                 13201 
                 acgccttcca agccatcgtc cacgccctcc aagccgacgc ccggcaccaa gccccccgag 
               
               
                   
               
               
                 13261 
                 tgcccagact ttgatcctcc cagacaggag aacgagactt ggtggctgtg cgactgcttc 
               
               
                   
               
               
                 13321 
                 atggccacgt gcaagtacaa caacacggtg gagatcgtga aggtggagtg tgagccgccg 
               
               
                   
               
               
                 13381 
                 cccatgccca cctgctccaa cggcctccaa cccgtgcgcg tcgaggaccc cgacggctgc 
               
               
                   
               
               
                 13441 
                 tgctggcact gggagtgcga ctgctactgc acgggctggg gcgacccgca ctatgtcacc 
               
               
                   
               
               
                 13501 
                 ttcgacggac tctactacag ctaccagggc aactgcacct acgtgctggt ggaggagatc 
               
               
                   
               
               
                 13561 
                 agcccctccg tggacaactt cggagtttac atcgacaact accactgcga tcccaacgac 
               
               
                   
               
               
                 13621 
                 aaggtgtcct gcccccgcac cctcatcgtg cgccacgaga cccaggaggt gctgatcaag 
               
               
                   
               
               
                 13681 
                 accgtgcata tgatgcccat gcaggtgcag gtgcaggtga acaggcaggc ggtggcactg 
               
               
                   
               
               
                 13741 
                 ccctacaaga agtacgggct ggaggtgtac cagtctggca tcaactacgt ggtggacatc 
               
               
                   
               
               
                 13801 
                 cccgagctgg gtgtcctcgt ctcctacaat ggcctgtcct tctccgtcag gctgccctac 
               
               
                   
               
               
                 13861 
                 caccggtttg gcaacaacac caagggccag tgtggcacct gcaccaacac cacctccgac 
               
               
                   
               
               
                 13921 
                 gactgcattc tgcccagcgg ggagatcgtc tccaactgtg aggctgcggc tgaccagtgg 
               
               
                   
               
               
                 13981 
                 ctggtgaacg acccctccaa gccacactgc ccccacagca gctccacgac caagcgcccg 
               
               
                   
               
               
                 14041 
                 gccgtcactg tgcccggggg cggtaaaacg accccacaca aggactgcac cccatctccc 
               
               
                   
               
               
                 14101 
                 ctctgccagc tcatcaagga cagcctgttt gcccagtgcc acgcactggt gcccccgcag 
               
               
                   
               
               
                 14161 
                 cactactacg atgcctgcgt gttcgacagc tgcttcatgc cgggctcgag cctggagtgc 
               
               
                   
               
               
                 14221 
                 gccagtctgc aggcctacgc agccctctgt gcccagcaga acatctgcct cgactggcgg 
               
               
                   
               
               
                 14281 
                 aaccacacgc atggggcctg cttggtggag tgcccatctc acagggagta ccaggcctgt 
               
               
                   
               
               
                 14341 
                 ggccctgcag aagagcccac gtgcaaatcc agctcctccc agcagaacaa cacagtcctg 
               
               
                   
               
               
                 14401 
                 gtggaaggct gcttctgtcc tgagggcacc atgaactacg ctcctggctt tgatgtctgc 
               
               
                   
               
               
                 14461 
                 gtgaagacct gcggctgtgt gggacctgac aatgtgccca gagagtttgg ggagcacttc 
               
               
                   
               
               
                 14521 
                 gagttcgact gcaagaactg tgtctgcctg gagggtggaa gtggcatcat ctgccaaccc 
               
               
                   
               
               
                 14581 
                 aagaggtgca gccagaagcc cgttacccac tgcgtggaag acggcaccta cctcgccacg 
               
               
                   
               
               
                 14641 
                 gaggtcaacc ctgccgacac ctgctgcaac attaccgtct gcaagtgcaa caccagcctg 
               
               
                   
               
               
                 14701 
                 tgcaaagaga agccctccgt gtgcccgctg ggattcgaag tgaagagcaa gatggtgcct 
               
               
                   
               
               
                 14761 
                 ggaaggtgct gtcctttcta ctggtgtgag tccaaggggg tgtgtgttca cgggaatgct 
               
               
                   
               
               
                 14821 
                 gagtaccagc ccggttctcc agtttattcc tccaagtgcc aggactgcgt gtgcacggac 
               
               
                   
               
               
                 14881 
                 aaggtggaca acaacaccct gctcaacgtc atcgcctgca cccacgtgcc ctgcaacacc 
               
               
                   
               
               
                 14941 
                 tcctgcagcc ctggcttcga actcatggag gcccccgggg agtgctgtaa gaagtgtgaa 
               
               
                   
               
               
                 15001 
                 cagacgcact gtatcatcaa acggcccgac aaccagcacg tcatcctgaa gcccggggac 
               
               
                   
               
               
                 15061 
                 ttcaagagcg acccgaagaa caactgcaca ttcttcagct gcgtgaagat ccacaaccag 
               
               
                   
               
               
                 15121 
                 ctcatctcgt ccgtctccaa catcacctgc cccaactttg atgccagcat ttgcatcccg 
               
               
                   
               
               
                 15181 
                 ggctccatca cattcatgcc caatggatgc tgcaagacct gcacccctcg caatgagacc 
               
               
                   
               
               
                 15241 
                 agggtgccct gctccaccgt ccccgtcacc acggaggttt cgtacgccgg ctgcaccaag 
               
               
                   
               
               
                 15301 
                 accgtcctca tgaatcattg ctccgggtcc tgcgggacat ttgtcatgta ctcggccaag 
               
               
                   
               
               
                 15361 
                 gcccaggccc tggaccacag ctgctcctgc tgcaaagagg agaaaaccag ccagcgtgag 
               
               
                   
               
               
                 15421 
                 gtggtcctga gctgccccaa tggcggctcg ctgacacaca cctacaccca catcgagagc 
               
               
                   
               
               
                 15481 
                 tgccagtgcc aggacaccgt ctgcgggctc cccaccggca cctcccgccg ggcccggcgc 
               
               
                   
               
               
                 15541 
                 tcccctaggc atctggggag cgggtgagcg gggtgggcac agcccccttc actgccctcg 
               
               
                   
               
               
                 15601 
                 acagctttac ctcccccgga ccctctgagc ctcctaagct cggcttcctc tcttcagata 
               
               
                   
               
               
                 15661 
                 tttattgtct gagtctttgt tcagtccttg ctttccaata ataaactcag ggggacatgc 
               
            
           
         
       
     
     By “NKX2-5 polypeptide” or “human NKX2-5 (hNKX2-5) polypeptide” is meant a protein or fragment thereof having at least 85% amino acid sequence identity to the sequence provided at NCBI Accession No. NP_004378.1 (isoform 1), NP_001159647.1 (isoform 2), or NP_001159648.1 (isoform 3) and having transcription factor activity. The amino acid sequence provided at NCBI Accession No. NP_004378.1 is shown below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 31) 
                   
               
            
           
           
               
               
               
            
               
                   1 
                 mfpspaltpt pfsvkdilnl eqqqrslaaa gelsarleat lapsscmlaa fkpeayagpe 
                   
               
               
                   
               
               
                  61 
                 aaapglpelr aelgrapspa kcasafpaap afypraysdp dpakdpraek kelcalqkav 
               
               
                   
               
               
                 121 
                 elekteadna erprarrrrk prvlfsqaqv yelerrfkgq rylsaperdq lasvlkltst 
               
               
                   
               
               
                 181 
                 qvkiwfqnrr ykckrqrqdq tlelvglppp pppparriav pvlvrdgkpc lgdsapyapa 
               
               
                   
               
               
                 241 
                 ygvglnpygy naypaypgyg gaacspgysc taaypagpsp agpataaann nfvnfgvgdl 
               
               
                   
               
               
                 301 
                 navqspgipq snsgvstlhg iraw 
               
            
           
         
       
     
     By “NKX2-5 polynucleotide” is meant a polynucleotide encoding a NKX2-5 polypeptide or fragment thereof. An exemplary NKX2-5 polynucleotide sequence is provided at NCBI Ref: NM_004387.3. The sequence provided at NCBI Ref: NM_004387.3 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 32) 
                   
               
            
           
           
               
               
               
            
               
                    1 
                 gctcctgtca tcgaggcccc tggcccaatg gcaggctgag tccccctcct ctggcctggt 
                   
               
               
                   
               
               
                   61 
                 cccgcctctc ctgccccttg tgctcagcgc tacctgctgc ccggacacat ccagagctgg 
               
               
                   
               
               
                  121 
                 ccgacgggtg cgcgggcggg cggcggcacc atgcagggaa gctgccaggg gccgtgggca 
               
               
                   
               
               
                  181 
                 gcgccgcttt ctgccgccca cctggcgctg tgagactggc gctgccacca tgttccccag 
               
               
                   
               
               
                  241 
                 ccctgctctc acgcccacgc ccttctcagt caaagacatc ctaaacctgg aacagcagca 
               
               
                   
               
               
                  301 
                 gcgcagcctg gctgccgccg gagagctctc tgcccgcctg gaggcgaccc tggcgccctc 
               
               
                   
               
               
                  361 
                 ctcctgcatg ctggccgcct tcaagccaga ggcctacgct gggcccgagg cggctgcgcc 
               
               
                   
               
               
                  421 
                 gggcctccca gagctgcgcg cagagctggg ccgcgcgcct tcaccggcca agtgtgcgtc 
               
               
                   
               
               
                  481 
                 tgcctttccc gccgcccccg ccttctatcc acgtgcctac agcgaccccg acccagccaa 
               
               
                   
               
               
                  541 
                 ggaccctaga gccgaaaaga aagagctgtg cgcgctgcag aaggcggtgg agctggagaa 
               
               
                   
               
               
                  601 
                 gacagaggcg gacaacgcgg agcggccccg ggcgcgacgg cggaggaagc cgcgcgtgct 
               
               
                   
               
               
                  661 
                 cttctcgcag gcgcaggtct atgagctgga gcggcgcttc aagcagcagc ggtacctgtc 
               
               
                   
               
               
                  721 
                 ggcccccgaa cgcgaccagc tggccagcgt gctgaaactc acgtccacgc aggtcaagat 
               
               
                   
               
               
                  781 
                 ctggttccag aaccggcgct acaagtgcaa gcggcagcgg caggaccaga ctctggagct 
               
               
                   
               
               
                  841 
                 ggtggggctg cccccgccgc cgccgccgcc tgcccgcagg atcgcggtgc cagtgctggt 
               
               
                   
               
               
                  901 
                 gcgcgatggc aagccatgcc taggggactc ggcgccctac gcgcctgcct acggcgtggg 
               
               
                   
               
               
                  961 
                 cctcaatccc tacggttata acgcctaccc cgcctatccg ggttacggcg gcgcggcctg 
               
               
                   
               
               
                 1021 
                 cagccctggc tacagctgca ctgccgctta ccccgccggg ccttccccag cgcagccggc 
               
               
                   
               
               
                 1081 
                 cactgccgcc gccaacaaca acttcgtgaa cttcggcgtc ggggacttga atgcggttca 
               
               
                   
               
               
                 1141 
                 gagccccggg attccgcaga gcaactcggg agtgtccacg ctgcatggta tccgagcctg 
               
               
                   
               
               
                 1201 
                 gtagggaagg gacccgcgtg gcgcgaccct gaccgatccc acctcaacag ctccctgact 
               
               
                   
               
               
                 1261 
                 ctcgggggga gaaggggctc ccaacatgac cctgagtccc ctggattttg cattcactcc 
               
               
                   
               
               
                 1321 
                 tgcggagacc taggaacttt ttctgtccca cgcgcgtttg ttcttgcgca cgggagagtt 
               
               
                   
               
               
                 1381 
                 tgtggcggcg attatgcagc gtgcaatgag tgatcctgca gcctggtgtc ttagctgtcc 
               
               
                   
               
               
                 1441 
                 ccccaggagt gccctccgag agtccatggg cacccccggt tggaactggg actgagctcg 
               
               
                   
               
               
                 1501 
                 ggcacgcagg gcctgagatc tggccgccca ttccgcgagc cagggccggg cgcccgggcc 
               
               
                   
               
               
                 1561 
                 tttgctatct cgccgtcgcc cgcccacgca cccacccgta tttatgtttt tacctattgc 
               
               
                   
               
               
                 1621 
                 tgtaagaaat gacgatcccc ttcccattaa agagagtgcg ttgaccccg 
               
            
           
         
       
     
     By “NEUROD1 polypeptide” is meant a protein or fragment thereof having at least 85% amino acid sequence identity to the sequence provided at NCBI Accession No. NP_002491.2 and having transcription factor activity. The amino acid sequence provided at NCBI Accession No. NP_002491.2 is shown below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 33) 
                   
               
            
           
           
               
               
               
            
               
                   1 
                 mtksysesgl mgepqpqgpp swtdeclssq deeheadkke ddleamnaee dslrnggeee 
                   
               
               
                   
               
               
                  61 
                 dededleeee eeeeedddqk pkrrgpkkkk mtkarlerfk lrrmkanare rnrmhglnaa 
               
               
                   
               
               
                 121 
                 ldnlrkvvpc ysktqklski etlrlaknyi walseilrsg kspdlvsfvq tlckglsgpt 
               
               
                   
               
               
                 181 
                 tnlvagclql nprtflpeqn qdmpphlpta sasfpvhpys yqspglpspp ygtmdsshvf 
               
               
                   
               
               
                 241 
                 hvkppphays aalepffesp ltdctspsfd gplspplsin gnfsfkheps aefeknyaft 
               
               
                   
               
               
                 301 
                 mhypaatlag aqshgsifsg taaprceipi dnimsfdshs hhervmsaql naifhd 
               
            
           
         
       
     
     By “NEUROD1 polynucleotide” is meant a polynucleotide encoding a NEUROD1 polypeptide or fragment thereof. An exemplary NEUROD1 polynucleotide sequence is provided at NCBI Ref: NM_002500.4. The sequence provided at NCBI Ref: NM_002500.4 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 34) 
                   
               
            
           
           
               
               
               
            
               
                    1 
                 ggggaggagg ggagaacggg gagcgcacag cctggacgcg tgcgcaggcg tcaggcgcat 
                   
               
               
                   
               
               
                   61 
                 agacctgcta gcccctcagc tagcggcccc gcccgcgctt agcatcacta actgggctat 
               
               
                   
               
               
                  121 
                 ataacctgag cgcccgcgcg gccacgacac gaggaattcg cccacgcagg aggcgcggcg 
               
               
                   
               
               
                  181 
                 tccggaggcc ccagggttat gagactatca ctgctcagga cctactaaca acaaaggaaa 
               
               
                   
               
               
                  241 
                 tcgaaacatg accaaatcgt acagcgagag tgggctgatg ggcgagcctc agccccaagg 
               
               
                   
               
               
                  301 
                 tcctccaagc tggacagacg agtgtctcag ttctcaggac gaggagcacg aggcagacaa 
               
               
                   
               
               
                  361 
                 gaaggaggac gacctcgaag ccatgaacgc agaggaggac tcactgagga acgggggaga 
               
               
                   
               
               
                  421 
                 ggaggaggac gaagatgagg acctggaaga ggaggaagaa gaggaagagg aggatgacga 
               
               
                   
               
               
                  481 
                 tcaaaagccc aagagacgcg gccccaaaaa gaagaagatg actaaggctc gcctggagcg 
               
               
                   
               
               
                  541 
                 ttttaaattg agacgcatga aggctaacgc ccgggagcgg aaccgcatgc acggactgaa 
               
               
                   
               
               
                  601 
                 cgcggcgcta gacaacctgc gcaaggtggt gccttgctat tctaagacgc agaagctgtc 
               
               
                   
               
               
                  661 
                 caaaatcgag actctgcgct tggccaagaa ctacatctgg gctctgtcgg agatcctgcg 
               
               
                   
               
               
                  721 
                 ctcaggcaaa agcccagacc tggtctcctt cgttcagacg ctttgcaagg gcttatccca 
               
               
                   
               
               
                  781 
                 acccaccacc aacctggttg cgggctgcct gcaactcaat cctcggactt ttctgcctga 
               
               
                   
               
               
                  841 
                 gcagaaccag gacatgcccc cccacctgcc gacggccagc gcttccttcc ctgtacaccc 
               
               
                   
               
               
                  901 
                 ctactcctac cagtcgcctg ggctgcccag tccgccttac ggtaccatgg acagctccca 
               
               
                   
               
               
                  961 
                 tgtcttccac gttaagcctc cgccgcacgc ctacagcgca gcgctggagc ccttctttga 
               
               
                   
               
               
                 1021 
                 aagccctctg actgattgca ccagcccttc ctttgatgga cccctcagcc cgccgctcag 
               
               
                   
               
               
                 1081 
                 catcaatggc aacttctctt tcaaacacga accgtccgcc gagtttgaga aaaattatgc 
               
               
                   
               
               
                 1141 
                 ctttaccatg cactatcctg cagcgacact ggcaggggcc caaagccacg gatcaatctt 
               
               
                   
               
               
                 1201 
                 ctcaggcacc gctgcccctc gctgcgagat ccccatagac aatattatgt ccttcgatag 
               
               
                   
               
               
                 1261 
                 ccattcacat catgagcgag tcatgagtgc ccagctcaat gccatatttc atgattagag 
               
               
                   
               
               
                 1321 
                 gcacgccagt ttcaccattt ccgggaaacg aacccactgt gcttacagtg actgtcgtgt 
               
               
                   
               
               
                 1381 
                 ttacaaaagg cagccctttg ggtactactg ctgcaaagtg caaatactcc aagcttcaag 
               
               
                   
               
               
                 1441 
                 tgatatatgt atttattgtc attactgcct ttggaagaaa caggggatca aagttcctgt 
               
               
                   
               
               
                 1501 
                 tcaccttatg tattattttc tatagctctt ctatttaaaa aataaaaaaa tacagtaaag 
               
               
                   
               
               
                 1561 
                 tttaaaaaat acaccacgaa tttggtgtgg ctgtattcag atcgtattaa ttatctgatc 
               
               
                   
               
               
                 1621 
                 gggataacaa aatcacaagc aataattagg atctatgcaa tttttaaact agtaatgggc 
               
               
                   
               
               
                 1681 
                 caattaaaat atatataaat atatattttt caaccagcat tttactactt gttacctttc 
               
               
                   
               
               
                 1741 
                 ccatgctgaa ttattttgtt gtgattttgt acagaatttt taatgacttt ttataatgtg 
               
               
                   
               
               
                 1801 
                 gatttcctat tttaaaacca tgcagcttca tcaattttta tacatatcag aaaagtagaa 
               
               
                   
               
               
                 1861 
                 ttatatctaa tttatacaaa ataatttaac taatttaaac cagcagaaaa gtgcttagaa 
               
               
                   
               
               
                 1921 
                 agttattgtg ttgccttagc acttctttcc tctccaattg taaaaaaaaa aaaaaaaaaa 
               
               
                   
               
               
                 1981 
                 aaaaaaaaaa aaaaattgca caatttgagc aattcatttc actttaaagt ctttccgtct 
               
               
                   
               
               
                 2041 
                 ccctaaaata aaaaccagaa tcataatttt caagagaaga aaaaattaag agatacattc 
               
               
                   
               
               
                 2101 
                 cctatcaaaa catatcaatt caacacatta cttgcacaag cttgtatata catattataa 
               
               
                   
               
               
                 2161 
                 ataaatgcca acataccctt ctttaaatca aaagctgctt gactatcaca tacaatttgc 
               
               
                   
               
               
                 2221 
                 actgttactt tttagtcttt tactcctttg cattccatga ttttacagag aatctgaagc 
               
               
                   
               
               
                 2281 
                 tattgatgtt tccagaaaat ataaatgcat gattttatac atagtcacaa aaatggtggt 
               
               
                   
               
               
                 2341 
                 ttgtcatata ttcatgtaat aaatctgagc ctaaatctaa tcaggttgtt aatgttggga 
               
               
                   
               
               
                 2401 
                 tttatatcta tagtagtcaa ttagtacagt agcttaaata aattcaaacc atttaattca 
               
               
                   
               
               
                 2461 
                 taattagaac aatagctatt gcatgtaaaa tgcagtccag aataagtgct gtttgagatg 
               
               
                   
               
               
                 2521 
                 tgatgctggt accactggaa tcgatctgta ctgtaatttt gtttgtaatc ctgtatatta 
               
               
                   
               
               
                 2581 
                 tggtgtaatg cacaatttag aaaacattca tccagttgca ataaaatagt attgaaagtg 
               
               
                   
               
               
                 2641 
                 agagcaattg ttgcatttct tcttaaaggg attctgtttt tatttttggg gaaagtagtt 
               
               
                   
               
               
                 2701 
                 gcttttttgc tgagttaaaa aatactaaac actatatgta gaataaaaga aaagaaaaaa 
               
               
                   
               
               
                 2761 
                 gtttaccttg gcatatgctc ttgtctgttt atcttgcaca gggagtcacc agttctatgt 
               
               
                   
               
               
                 2821 
                 agataatgaa aagacctaac tgatatttca ttatttggaa tatgggactg gacggcagta 
               
               
                   
               
               
                 2881 
                 caaacagtgt gtttttttct ttgttttaag tggcttagcc tttaggtttt ttatttccat 
               
               
                   
               
               
                 2941 
                 ttttaaaaat gattgttaca tgttttcttc tatttctttt tttaaaaggt ggattttaat 
               
               
                   
               
               
                 3001 
                 aa 
               
            
           
         
       
     
     By “NKX6-1 polypeptide” is meant a protein or fragment thereof having at least 85% amino acid sequence identity to the sequence provided at NCBI Accession No. NP_006159.2 and having transcription factor activity. The amino acid sequence provided at NCBI Accession No. NP_006159.2 is shown below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 35) 
                   
               
            
           
           
               
               
               
            
               
                   1 
                 mlavgamegt rqsafllssp plaalhsmae mktplypaay pplpagppss sssssssssp 
                   
               
               
                   
               
               
                  61 
                 spplgthnpg glkppatggl sslgsppqql saatphgind ilsrpsmpva sgaalpsasp 
               
               
                   
               
               
                 121 
                 sgssssssss asassasaaa aaaaaaaaaa sspagllagl prfsslsppp pppglyfsps 
               
               
                   
               
               
                 181 
                 aaavaavgry pkplaelpgr tpifwpgvmq sppwrdarla ctphqgsill dkdgkrkhtr 
               
               
                   
               
               
                 241 
                 ptfsgqqifa lektfeqtky lagperarla yslgmtesqv kvwfqnrrtk wrkkhaaema 
               
               
                   
               
               
                 301 
                 takkkqdset erlkgasene eedddynkpl dpnsddekit qllkkhksss ggggglllha 
               
               
                   
               
               
                 361 
                 sepesss 
               
            
           
         
       
     
     By “NKX6-1 polynucleotide” is meant a polynucleotide encoding a NKX6-1 polypeptide or fragment thereof. An exemplary NKX6-1 polynucleotide sequence is provided at NCBI Ref: NM_006168.2. The sequence provided at NCBI Ref: NM_006168.2 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 36) 
                   
               
            
           
           
               
               
               
            
               
                    1 
                 cgtgggatgt tagcggtggg ggcaatggag ggcacccggc agagcgcatt cctgctcagc 
                   
               
               
                   
               
               
                   61 
                 agccctcccc tggccgccct gcacagcatg gccgagatga agaccccgct gtaccctgcc 
               
               
                   
               
               
                  121 
                 gcgtatcccc cgctgcctgc cggccccccc tcctcctcgt cctcgtcgtc gtcctcctcg 
               
               
                   
               
               
                  181 
                 tcgccctccc cgcctctggg cacccacaac ccaggcggcc tgaagccccc ggccacgggg 
               
               
                   
               
               
                  241 
                 gggctctcat ccctcggcag ccccccgcag cagctctcgg ccgccacccc acacggcatc 
               
               
                   
               
               
                  301 
                 aacgatatcc tgagccggcc ctccatgccc gtggcctcgg gggccgccct gccctccgcc 
               
               
                   
               
               
                  361 
                 tcgccctccg gttcctcctc ctcctcttcc tcgtccgcct ctgcctcctc cgcctctgcc 
               
               
                   
               
               
                  421 
                 gccgccgcgg ctgctgccgc ggccgcagcc gccgcctcat ccccggcggg gctgctggcc 
               
               
                   
               
               
                  481 
                 ggactgccac gctttagcag cctgagcccg ccgccgccgc cgcccgggct ctacttcagc 
               
               
                   
               
               
                  541 
                 cccagcgccg cggccgtggc cgccgtgggc cggtacccca agccgctggc tgagctgcct 
               
               
                   
               
               
                  601 
                 ggccggacgc ccatcttctg gcccggagtg atgcagagcc cgccctggag ggacgcacgc 
               
               
                   
               
               
                  661 
                 ctggcctgta cccctcatca aggatccatt ttgttggaca aagacgggaa gagaaaacac 
               
               
                   
               
               
                  721 
                 acgagaccca ctttttccgg acagcagatc ttcgccctgg agaagacttt cgaacaaaca 
               
               
                   
               
               
                  781 
                 aaatacttgg cggggcccga gagggctcgt ttggcctatt cgttggggat gacagagagt 
               
               
                   
               
               
                  841 
                 caggtcaagg tctggttcca gaaccgccgg accaagtgga ggaagaagca cgctgccgag 
               
               
                   
               
               
                  901 
                 atggccacgg ccaagaagaa gcaggactcg gagacagagc gcctcaaggg ggcctcggag 
               
               
                   
               
               
                  961 
                 aacgaggaag aggacgacga ctacaataag cctctggatc ccaactcgga cgacgagaaa 
               
               
                   
               
               
                 1021 
                 atcacgcagc tgttgaagaa gcacaagtcc agcagcggcg gcggcggcgg cctcctactg 
               
               
                   
               
               
                 1081 
                 cacgcgtccg agccggagag ctcatcctga acgccg 
               
            
           
         
       
     
     By “NDUFA4 polypeptide” is meant a protein or fragment thereof having at least 85% amino acid sequence identity to the sequence provided at NCBI Accession No. NP_002480.1 and having NADH dehydrogenase activity and oxidoreductase activity. The amino acid sequence provided at NCBI Accession No. NP_002480.1 is shown below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 37) 
                   
               
            
           
           
               
               
               
            
               
                   1 
                 maaelamgae lpssplaiey vndfdlmkfe vkkeppeaer fchrlppgsl sstplstpcs 
                   
               
               
                   
               
               
                  61 
                 svpsspsfca pspgtggggg agggggssqa ggapgppsgg pgavggtsgk paledlywms 
               
               
                   
               
               
                 121 
                 gyqhhlnpea lnltpedave aligsghhga hhgahhpaaa aayeafrgpg faggggaddm 
               
               
                   
               
               
                 181 
                 gaghhhgahh aahhhhaahh hhhhhhhhgg aghgggaghh vrleerfsdd qlvsmsvrel 
               
               
                   
               
               
                 241 
                 nrqlrgfske evirlkqkrr tlknrgyaqs crfkrvqqrh ilesekcqlq sqveqlklev 
               
               
                   
               
               
                 301 
                 grlakerdly kekyeklagr ggpgsaggag fprepsppqa gpggakgtad ffl 
               
            
           
         
       
     
     By “NDUFA4 polynucleotide” is meant a polynucleotide encoding a NDUFA4 polypeptide or fragment thereof. An exemplary NDUFA4 polynucleotide sequence is provided at NCBI Ref: NM_002489.3. The sequence provided at NCBI Ref: NM_002489.3 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 38) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 gggtccttca ggtaggaggt cctgggtgac tttggaagtc cgtagtgtct cattgcagat 
                   
               
               
                   
               
               
                 61 
                 aatttttagc ttagggcctg gtggctaggt cggttctctc ctttccagtc ggagacctct 
               
               
                   
               
               
                 121 
                 gccgcaaaca tgctccgcca gatcatcggt caggccaaga agcatccgag cttgatcccc 
               
               
                   
               
               
                 181 
                 ctctttgtat ttattggaac tggagctact ggagcaacac tgtatctctt gcgtctggca 
               
               
                   
               
               
                 241 
                 ttgttcaatc cagatgtttg ttgggacaga aataacccag agccctggaa caaactgggt 
               
               
                   
               
               
                 301 
                 cccaatgatc aatacaagtt ctactcagtg aatgtggatt acagcaagct gaagaaggaa 
               
               
                   
               
               
                 361 
                 cgtccagatt tctaaatgaa atgtttcact ataacgctgc tttagaatga aggtcttcca 
               
               
                   
               
               
                 421 
                 gaagccacat ccgcacaatt ttccacttaa ccaggaaata tttctcctct aaatgcatga 
               
               
                   
               
               
                 481 
                 aatcatgttg gagatctcta ttgtaatctc tattggagat tacaatgatt aaatcaataa 
               
               
                   
               
               
                 541 
                 ataactgaaa cttgatatgt gtcacttttt tatgctgaaa gtatgctctg aactttagag 
               
               
                   
               
               
                 601 
                 tataggaaat taactattag aatttaaaga atttcttgaa tttctgtagt ttgaaaatac 
               
               
                   
               
               
                 661 
                 gactttaagc tgctttagta aaacacttcc attttgtgta tagactgttg gtaacttcac 
               
               
                   
               
               
                 721 
                 tagagcatac ataacaactg gaactggaaa ttatacaaaa gtaaattggg aaggatactc 
               
               
                   
               
               
                 781 
                 cagcatctga cactggcaaa atggaaacct ttgagtttct cttactggct gttgaagtgt 
               
               
                   
               
               
                 841 
                 gtgcagtttt taacaatggt ttttacttgg catctctttg ttgtgatttt caaggttata 
               
               
                   
               
               
                 901 
                 agttgctttg gtcctaggat tgaagttgaa atctgagttt atcagtgcta accatggtgc 
               
               
                   
               
               
                 961 
                 tagtagtcaa gagatcttga gaattttggc tgctgagtct tggtgcaggg tgcaggtttt 
               
               
                   
               
               
                 1021 
                 cttttctttt ttcttttttt tttttttgag atagtctctg tcacccaggc tggagtgcag 
               
               
                   
               
               
                 1081 
                 tggtacaaac atggatcact gcagcctcta cctcccgggc ttaagtgatc ctcctgcctc 
               
               
                   
               
               
                 1141 
                 agcccctaag tagccgggac tacaggtatg tgccaccatg cccagttaat ttttgtaatt 
               
               
                   
               
               
                 1201 
                 ttttttagag acagggtttt gccatgttgc ccaggctggt ctcaaactct tgagctcaag 
               
               
                   
               
               
                 1261 
                 cgatccattc tcctcagcct cccagggtgc tgggattaca ggcgtgagcc attgcgctta 
               
               
                   
               
               
                 1321 
                 gccatggtgc aggttttcaa aggccaggaa gtatattcat aattttaaga tggggaatat 
               
               
                   
               
               
                 1381 
                 agcaagtttt cacataggtg tgtgtaagtc atcacatcat agaaacttga ggaattcagt 
               
               
                   
               
               
                 1441 
                 gacattaatt ttggattttc atacgtaagt atacaattaa atgtttacag ggtagtagaa 
               
               
                   
               
               
                 1501 
                 gcacatttta aatgtcagga actgaactaa gtatttgaat tacgtggatt atctcaaaaa 
               
               
                   
               
               
                 1561 
                 ttttgaaatt gttaaacgag ttgaattact tgaattcatt ctgttagtca aatggtggat 
               
               
                   
               
               
                 1621 
                 atttacaccc atgtagtttt gaatttagag tgtgtagagt gttttcagtt accagactcc 
               
               
                   
               
               
                 1681 
                 atgcttttac ctcctatgtg tcaggtataa tttgaacctc taagaacagg gtttctcaac 
               
               
                   
               
               
                 1741 
                 cttgccactg ttgactattt ctgaaagaca gtttggttta gcagaccatc ccatgcgctt 
               
               
                   
               
               
                 1801 
                 tagcttgttt agtagctaac ttgggctctg ccactacaga caaaaagcac tctttccctc 
               
               
                   
               
               
                 1861 
                 caattcccac aggctatgag aagaatggag acattaccaa atgtccattg gtgggcaaaa 
               
               
                   
               
               
                 1921 
                 ttgcttcatt cctacctctg ttgagaatta ctctagatcc tttggcacaa attacctcaa 
               
               
                   
               
               
                 1981 
                 agtttaaaat tgtgtaaaca aacagtgtgt catgtaattg aaaaacatta agcaactcca 
               
               
                   
               
               
                 2041 
                 aataaatgct acattaag 
               
            
           
         
       
     
     As used herein, “obtaining” as in “obtaining an agent” includes synthesizing, purchasing, or otherwise acquiring the agent. 
     By “organ” is meant a collection of cells that perform a biological function. In one embodiment, an organ includes, but is not limited to, bladder, brain, nervous tissue, glial tissue, esophagus, fallopian tube, heart, pancreas, intestines, gallbladder, kidney, liver, lung, ovaries, prostate, spinal cord, spleen, stomach, testes, thymus, thyroid, trachea, urogenital tract, ureter, urethra, uterus, breast, skeletal muscle, skin, bone, and cartilage. The biological function of an organ can be assayed using standard methods known to the skilled artisan. 
     By “organoid” is meant an in vitro generated body that mimics organ structure and function. “Organoid” and “mini organ” are used interchangeably herein. A “pancreatic islet organoid” is an in vitro generated cell cluster that mimics structure and function of a pancreatic islet. Exemplary functions of a pancreatic islet include, without limitation, glucose-stimulated insulin secretion (GSIS), potassium chloride (KCl)-stimulated insulin secretion, GLP-1 stimulated insulin secretion, somatostatin secretion, or glucagon secretion. “Pancreatic islet organoid” and “mini pancreatic islet” are used interchangeably herein. A “pancreatic organoid” is an in vitro generated body that mimics structure and function of a pancreas. Exemplary functions of a pancreas include, without limitation, endocrine secretion of hormones, such as glucose and glucagon, that regulate glucose metabolism and blood glucose concentration, and exocrine secretion of digestive enzymes that help break down carbohydrates, proteins, and lipids. “Pancreatic organoid” and “mini pancreas” are used interchangeably herein. 
     By “PAX4 polypeptide” is meant a protein or fragment thereof having at least 85% amino acid sequence identity to the sequence provided at NCBI Accession No. NP_006184.2 and having transcription factor activity. The amino acid sequence provided at NCBI Accession No. NP_006184.2 is shown below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 39) 
                   
               
            
           
           
               
               
               
            
               
                   1 
                 mnqlgglfvn grplpldtrq qivrlavsgm rpcdisrilk vsngcvskil gryyrtgvle 
                   
               
               
                   
               
               
                  61 
                 pkgiggskpr latppvvari aqlkgecpal faweiqrqlc aeglctqdkt psvssinrvl 
               
               
                   
               
               
                 121 
                 ralqedqglp ctrlrspavl apavltphsg setprgthpg tghrnrtifs psqaealeke 
               
               
                   
               
               
                 181 
                 fqrgqypdsv argklatats lpedtvrvwf snrrakwrrq eklkwemqlp gasqgltvpr 
               
               
                   
               
               
                 241 
                 vapgiisaqq spgsvptaal paleplgpsc yqlcwatape rclsdtppka clkpcwghlp 
               
               
                   
               
               
                 301 
                 pqpnsldsgl lclpcpsshc hlaslsgsqa llwpgcplly gle 
               
            
           
         
       
     
     By “PAX4 polynucleotide” is meant a polynucleotide encoding a PAX4 polypeptide or fragment thereof. An exemplary PAX4 polynucleotide sequence is provided at NCBI Ref: NM_006193.2. The sequence provided at NCBI Ref: NM_006193.2 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 40) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 caaagactca cccgtgagcc agctctcaaa gaaagcagct tgcgttgaca gcctgggggc 
                   
               
               
                   
               
               
                 61 
                 agcaaggatg cagtctccca ggagaggatg cactcggtgg tgggaagcca ggctggaggg 
               
               
                   
               
               
                 121 
                 gcctgagtga ccctctccac aggcgggcag ggcagtggga gaggtggtgt gtggatacct 
               
               
                   
               
               
                 181 
                 ctgtctcacg cccagggatc agcagcatga accagcttgg ggggctcttt gtgaatggcc 
               
               
                   
               
               
                 241 
                 ggcccctgcc tctggatacc cggcagcaga ttgtgcggct agcagtcagt ggaatgcggc 
               
               
                   
               
               
                 301 
                 cctgtgacat ctcacggatc cttaaggtat ctaatggctg tgtgagcaag atcctagggc 
               
               
                   
               
               
                 361 
                 gttactaccg cacaggtgtc ttggagccaa agggcattgg gggaagcaag ccacggctgg 
               
               
                   
               
               
                 421 
                 ctacaccccc tgtggtggct cgaattgccc agctgaaggg tgagtgtcca gccctctttg 
               
               
                   
               
               
                 481 
                 cctgggaaat ccaacgccag ctttgtgctg aagggctttg cacccaggac aagactccca 
               
               
                   
               
               
                 541 
                 gtgtctcctc catcaaccga gtcctgcggg cattacagga ggaccaggga ctaccgtgca 
               
               
                   
               
               
                 601 
                 cacggctcag gtcaccagct gttttggctc cagctgtcct cactccccat agtggctctg 
               
               
                   
               
               
                 661 
                 agactccccg gggtacccac ccagggaccg gccaccggaa tcggactatc ttctccccaa 
               
               
                   
               
               
                 721 
                 gccaagcaga ggcactggag aaagagttcc agcgtgggca gtatcctgat tcagtggccc 
               
               
                   
               
               
                 781 
                 gtggaaagct ggctactgcc acctctctgc ctgaggacac ggtgagggtc tggttttcca 
               
               
                   
               
               
                 841 
                 acagaagagc caaatggcgt cggcaagaga agctcaagtg ggaaatgcag ctgccaggtg 
               
               
                   
               
               
                 901 
                 cttcccaggg gctgactgta ccaagggttg ccccaggaat catctctgca cagcagtccc 
               
               
                   
               
               
                 961 
                 ctggcagtgt gcccacagca gccctgcctg ccctggaacc actgggtccc tcctgctatc 
               
               
                   
               
               
                 1021 
                 agctgtgctg ggcaacagca ccagaaaggt gtctgagtga caccccacct aaagcctgtc 
               
               
                   
               
               
                 1081 
                 tcaagccctg ctggggccac ttgcccccac agccgaattc cctggactca ggactgcttt 
               
               
                   
               
               
                 1141 
                 gccttccttg cccttcctcc cactgtcacc tggccagtct tagtggctct caggccctgc 
               
               
                   
               
               
                 1201 
                 tctggcctgg ctgcccacta ctgtatggct tggaatgagg caggagtggg aaggagatgg 
               
               
                   
               
               
                 1261 
                 catagagaag atctaatacc atcctgccca ttgtccttac cgtcctgccc atacagactg 
               
               
                   
               
               
                 1321 
                 tggctccttc ctccttcctg tgattgctcc ctcctgtgtg gacgttgcct ggccctgcct 
               
               
                   
               
               
                 1381 
                 cgatgcctct ctggcgcatc acctgattgg aggggctggt aaagcaacac ccacccactt 
               
               
                   
               
               
                 1441 
                 ctcacactag ccttaagagg cctccactca gcagtaataa aagctgtttt tattagcagt 
               
               
                   
               
               
                 1501 
                 agttctgttg tccatcatgt tttccctatg agcaccccta tgcccactct aatattcaac 
               
               
                   
               
               
                 1561 
                 aattatagac aatttgccct atcatttatt tacatctatg tatctaccat ctaatctatg 
               
               
                   
               
               
                 1621 
                 catgtatgta ggcaatacat gtatctaaac aatgtatttg tcaatgcatc aatttaccta 
               
               
                   
               
               
                 1681 
                 ctctatgtat gcatctatat gtgtattatg tatgcgtgca tgcgtgcgcg cacacacaca 
               
               
                   
               
               
                 1741 
                 cacacacaca cacactgaca ttatatcatg gcattttatt cctaaatctt ccagcatgca 
               
               
                   
               
               
                 1801 
                 tccccaaaaa acaagaaact tgtcttacat aatcacaata atatatccac atctaagaaa 
               
               
                   
               
               
                 1861 
                 atttactgta acttcttaat ctaagaaaat tatgtatttt tgtcatatgt attttgtcat 
               
               
                   
               
               
                 1921 
                 atgtattttg tatttgcata tgtattttgt atttgcatat gtatttttgt catagcagca 
               
               
                   
               
               
                 1981 
                 aacagagtga aatgccattt ttcatattct 
               
            
           
         
       
     
     By “PAX6 polypeptide” is meant a protein or fragment thereof having at least 85% amino acid sequence identity to the sequence provided at NCBI Accession No. NP_001297090.1 and having transcription factor activity. The amino acid sequence provided at NCBI Accession No. NP_001297090.1 is shown below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 41) 
                   
               
            
           
           
               
               
               
            
               
                   1 
                 mgadgmydkl rmlngqtgsw gtrpgwypgt svpgqptqdg cqqqegggen tnsissnged 
                   
               
               
                   
               
               
                  61 
                 sdeaqmrlql krklqrnrts ftqeqieale keferthypd vfarerlaak idlpeariqv 
               
               
                   
               
               
                 121 
                 wfsnrrakwr reeklrnqrr qasntpship isssfstsvy qpipqpttpv ssftsgsmlg 
               
               
                   
               
               
                 181 
                 rtdtaltnty salppmpsft mannlpmqpp vpsqtssysc mlptspsvng rsydtytpph 
               
               
                   
               
               
                 241 
                 mqthmnsqpm gtsgttstgl ispgvsvpvq vpgsepdmsq ywprlq 
               
            
           
         
       
     
     By “PAX6 polynucleotide” is meant a polynucleotide encoding a PAX6 polypeptide or fragment thereof. An exemplary PAX6 polynucleotide sequence is provided at NCBI Ref: NM_001310161.1. The sequence provided at NCBI Ref: NM_001310161.1 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 42) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 cttttcaatt agccttccat gcatgatccg gagcgacttc cgcctatttc cagaaattaa 
                   
               
               
                   
               
               
                 61 
                 gctcaaactt gacgtgcagc tagttttatt ttaaagacaa atgtcagaga ggctcatcat 
               
               
                   
               
               
                 121 
                 attttccccc ctcttctata tttggagctt atttattgct aagaagctca ggctcctggc 
               
               
                   
               
               
                 181 
                 gtcaatttat cagtaggctc caaggagaag agaggagagg agaggagagc tgaacaggga 
               
               
                   
               
               
                 241 
                 gccacgtctt ttcctgggag ggctgctatc taagtcgggg ctgcaggtca cagcggagtg 
               
               
                   
               
               
                 301 
                 aatcagctcg gtggtgtctt tgtcaacggg cggccactgc cggactccac ccggcagaag 
               
               
                   
               
               
                 361 
                 attgtagagc tagctcacag cggggcccgg ccgtgcgaca tttcccgaat tctgcagacc 
               
               
                   
               
               
                 421 
                 catgcagatg caaaagtcca agtgctggac aatcaaaacg tgtccaacgg atgtgtgagt 
               
               
                   
               
               
                 481 
                 aaaattctgg gcaggtatta cgagactggc tccatcagac ccagggcaat cggtggtagt 
               
               
                   
               
               
                 541 
                 aaaccgagag tagcgactcc agaagttgta agcaaaatag cccagtataa gcgggagtgc 
               
               
                   
               
               
                 601 
                 ccgtccatct ttgcttggga aatccgagac agattactgt ccgagggggt ctgtaccaac 
               
               
                   
               
               
                 661 
                 gataacatac caagcgtgtc atcaataaac agagttcttc gcaacctggc tagcgaaaag 
               
               
                   
               
               
                 721 
                 caacagatgg gcgcagacgg catgtatgat aaactaagga tgttgaacgg gcagaccgga 
               
               
                   
               
               
                 781 
                 agctggggca cccgccctgg ttggtatccg gggacttcgg tgccagggca acctacgcaa 
               
               
                   
               
               
                 841 
                 gatggctgcc agcaacagga aggaggggga gagaatacca actccatcag ttccaacgga 
               
               
                   
               
               
                 901 
                 gaagattcag atgaggctca aatgcgactt cagctgaagc ggaagctgca aagaaataga 
               
               
                   
               
               
                 961 
                 acatccttta cccaagagca aattgaggcc ctggagaaag agtttgagag aacccattat 
               
               
                   
               
               
                 1021 
                 ccagatgtgt ttgcccgaga aagactagca gccaaaatag atctacctga agcaagaata 
               
               
                   
               
               
                 1081 
                 caggtatggt tttctaatcg aagggccaaa tggagaagag aagaaaaact gaggaatcag 
               
               
                   
               
               
                 1141 
                 agaagacagg ccagcaacac acctagtcat attcctatca gcagtagttt cagcaccagt 
               
               
                   
               
               
                 1201 
                 gtctaccaac caattccaca acccaccaca ccggtttcct ccttcacatc tggctccatg 
               
               
                   
               
               
                 1261 
                 ttgggccgaa cagacacagc cctcacaaac acctacagcg ctctgccgcc tatgcccagc 
               
               
                   
               
               
                 1321 
                 ttcaccatgg caaataacct gcctatgcaa cccccagtcc ccagccagac ctcctcatac 
               
               
                   
               
               
                 1381 
                 tcctgcatgc tgcccaccag cccttcggtg aatgggcgga gttatgatac ctacaccccc 
               
               
                   
               
               
                 1441 
                 ccacatatgc agacacacat gaacagtcag ccaatgggca cctcgggcac cacttcaaca 
               
               
                   
               
               
                 1501 
                 ggactcattt cccctggtgt gtcagttcca gttcaagttc ccggaagtga acctgatatg 
               
               
                   
               
               
                 1561 
                 tctcaatact ggccaagatt acagtaaaaa aaaaaaaaaa aaaaaaaagg aaaggaaata 
               
               
                   
               
               
                 1621 
                 ttgtgttaat tcagtcagtg actatgggga cacaacagtt gagctttcag gaaagaaaga 
               
               
                   
               
               
                 1681 
                 aaaatggctg ttagagccgc ttcagttcta caattgtgtc ctgtattgta ccactgggga 
               
               
                   
               
               
                 1741 
                 aggaatggac ttgaaacaag gacctttgta tacagaaggc acgatatcag ttggaacaaa 
               
               
                   
               
               
                 1801 
                 tcttcatttt ggtatccaaa cttttattca ttttggtgta ttatttgtaa atgggcattt 
               
               
                   
               
               
                 1861 
                 gtatgttata atgaaaaaaa gaacaatgta gactggatgg atgtttgatc tgtgttggtc 
               
               
                   
               
               
                 1921 
                 atgaagttgt tttttttttt tttaaaaaga aaaccatgat caacaagctt tgccacgaat 
               
               
                   
               
               
                 1981 
                 ttaagagttt tatcaagata tatcgaatac ttctacccat ctgttcatag tttatggact 
               
               
                   
               
               
                 2041 
                 gatgttccaa gtttgtatca ttcctttgca tataattaaa cctggaacaa catgcactag 
               
               
                   
               
               
                 2101 
                 atttatgtca gaaatatctg ttggttttcc aaaggttgtt aacagatgaa gtttatgtgc 
               
               
                   
               
               
                 2161 
                 aaaaaagggt aagatataaa ttcaaggaag aaaaaaagtt gatagctaaa aggtagagtg 
               
               
                   
               
               
                 2221 
                 tgtcttcgat ataatccaat ttgttttatg tcaaaatgta agtatttgtc ttccctagaa 
               
               
                   
               
               
                 2281 
                 atcctcagaa tgatttctat aataaagtta atttcattta tatttgacaa gaatatagat 
               
               
                   
               
               
                 2341 
                 gttttataca cattttcatg caatcatacg tttctttttt ggccagcaaa agttaattgt 
               
               
                   
               
               
                 2401 
                 tcttagatat agttgtatta ctgttcacgg tccaatcatt ttgtgcatct agagttcatt 
               
               
                   
               
               
                 2461 
                 cctaatcaat taaaagtgct tgcaagagtt ttaaacttaa gtgttttgaa gttgttcaca 
               
               
                   
               
               
                 2521 
                 actacatatc aaaattaacc attgttgatt gtaaaaaacc atgccaaagc ctttgtattt 
               
               
                   
               
               
                 2581 
                 cctttattat acagttttct ttttaacctt atagtgtggt gttacaaatt ttatttccat 
               
               
                   
               
               
                 2641 
                 gttagatcaa cattctaaac caatggttac tttcacacac actctgtttt acatcctgat 
               
               
                   
               
               
                 2701 
                 gatccttaaa aaataatcct tatagatacc ataaatcaaa aacgtgttag aaaaaaattc 
               
               
                   
               
               
                 2761 
                 cacttacagc agggtgtaga tctgtgccca tttataccca caacatatat acaaaatggt 
               
               
                   
               
               
                 2821 
                 aacatttccc agttagccat ttaattctaa agctcaaagt ctagaaataa tttaaaaatg 
               
               
                   
               
               
                 2881 
                 caacaagcga ttagctagga attgtttttt gaattaggac tggcattttc aatctgggca 
               
               
                   
               
               
                 2941 
                 gatttccatt gtcagcctat ttcaacaatg atttcactga agtatattca aaagtagatt 
               
               
                   
               
               
                 3001 
                 tcttaaagga gactttctga aagctgttgc ctttttcaaa taggccctct cccttttctg 
               
               
                   
               
               
                 3061 
                 tctccctccc ctttgcacaa gaggcatcat ttcccattga accactacag ctgttcccat 
               
               
                   
               
               
                 3121 
                 ttgaatcttg ctttctgtgc ggttgtggat ggttggaggg tggagggggg atgttgcatg 
               
               
                   
               
               
                 3181 
                 tcaaggaata atgagcacag acacatcaac agacaacaac aaagcagact gtgactggcc 
               
               
                   
               
               
                 3241 
                 ggtgggaatt aaaggccttc agtcattggc agcttaagcc aaacattccc aaatctatga 
               
               
                   
               
               
                 3301 
                 agcagggccc attgttggtc agttgttatt tgcaatgaag cacagttctg atcatgttta 
               
               
                   
               
               
                 3361 
                 aagtggaggc acgcagggca ggagtgcttg agcccaagca aaggatggaa aaaaataagc 
               
               
                   
               
               
                 3421 
                 ctttgttggg taaaaaagga ctgtctgaga ctttcatttg ttctgtgcaa catataagtc 
               
               
                   
               
               
                 3481 
                 aatacagata agtcttcctc tgcaaacttc actaaaaagc ctgggggttc tggcagtcta 
               
               
                   
               
               
                 3541 
                 gattaaaatg cttgcacatg cagaaacctc tggggacaaa gacacacttc cactgaatta 
               
               
                   
               
               
                 3601 
                 tactctgctt taaaaaaatc cccaaaagca aatgatcaga aatgtagaaa ttaatggaag 
               
               
                   
               
               
                 3661 
                 gatttaaaca tgaccttctc gttcaatatc tactgttttt tagttaagga attacttgtg 
               
               
                   
               
               
                 3721 
                 aacagataat tgagattcat tgctccggca tgaaatatac taataatttt attccaccag 
               
               
                   
               
               
                 3781 
                 agttgctgca catttggaga caccttccta agttgcagtt tttgtatgtg tgcatgtagt 
               
               
                   
               
               
                 3841 
                 tttgttcagt gtcagcctgc actgcacagc agcacatttc tgcaggggag tgagcacaca 
               
               
                   
               
               
                 3901 
                 tacgcactgt tggtacaatt gccggtgcag acatttctac ctcctgacat tttgcagcct 
               
               
                   
               
               
                 3961 
                 acattccctg agggctgtgt gctgagggaa ctgtcagaga agggctatgt gggagtgcat 
               
               
                   
               
               
                 4021 
                 gccacagctg ctggctggct tacttcttcc ttctcgctgg ctgtaatttc caccacggtc 
               
               
                   
               
               
                 4081 
                 aggcagccag ttccggccca cggttctgtt gtgtagacag cagagacttt ggagacccgg 
               
               
                   
               
               
                 4141 
                 atgtcgcacg ccaggtgcaa gaggtgggaa tgggagaaaa ggagtgacgt gggagcggag 
               
               
                   
               
               
                 4201 
                 ggtctgtatg tgtgcacttg ggcacgtata tgtgtgctct gaaggtcagg attgccaggg 
               
               
                   
               
               
                 4261 
                 caaagtagca cagtctggta tagtctgaag aagcggctgc tcagctgcag aagccctctg 
               
               
                   
               
               
                 4321 
                 gtccggcagg atgggaacgg ctgccttgcc ttctgcccac accctaggga catgagctgt 
               
               
                   
               
               
                 4381 
                 ccttccaaac agagctccag gcactctctt ggggacagca tggcaggctc tgtgtggtag 
               
               
                   
               
               
                 4441 
                 cagtgcctgg gagttggcct tttactcatt gttgaaataa tttttgttta ttatttattt 
               
               
                   
               
               
                 4501 
                 aacgatacat atatttatat atttatcaat ggggtatctg cagggatgtt ttgacaccat 
               
               
                   
               
               
                 4561 
                 cttccaggat ggagattatt tgtgaagact tcagtagaat cccaggacta aacgtctaaa 
               
               
                   
               
               
                 4621 
                 ttttttctcc aaacttgact gacttgggaa aaccaggtga atagaataag agctgaatgt 
               
               
                   
               
               
                 4681 
                 tttaagtaat aaacgttcaa actgctctaa gtaaaaaaat gcattttact gcaatgaatt 
               
               
                   
               
               
                 4741 
                 tctagaatat ttttccccca aagctatgcc tcctaaccct taaatggtga acaactggtt 
               
               
                   
               
               
                 4801 
                 tcttgctaca gctcactgcc atttcttctt actatcatca ctaggtttcc taagattcac 
               
               
                   
               
               
                 4861 
                 tcatacagta ttatttgaag attcagcttt gttctgtgaa tgtcatctta ggattgtgtc 
               
               
                   
               
               
                 4921 
                 tatattcttt tgcttatttc tttttactct gggcctctca tactagtaag attttaaaaa 
               
               
                   
               
               
                 4981 
                 gccttttctt ctctgtatgt ttggctcacc aaggcgaaat atatattctt ctctttttca 
               
               
                   
               
               
                 5041 
                 tttctcaaga ataaacctca tctgcttttt tgtttttctg tgttttggct tggtactgaa 
               
               
                   
               
               
                 5101 
                 tgactcaact gctcggtttt aaagttcaaa gtgtaagtac ttagggttag tactgcttat 
               
               
                   
               
               
                 5161 
                 ttcaataatg ttgacggtga ctatctttgg aaagcagtaa catgctgtct tagaaatgac 
               
               
                   
               
               
                 5221 
                 attaataatg ggcttaaaca aatgaatagg ggggtccccc cactctcctt ttgtatgcct 
               
               
                   
               
               
                 5281 
                 atgtgtgtct gatttgttaa aagatggaca gggaattgat tgcagagtgt cgcttccttc 
               
               
                   
               
               
                 5341 
                 taaagtagtt ttattttgtc tactgttagt atttaaagat cctggaggtg gacataagga 
               
               
                   
               
               
                 5401 
                 ataaatggaa gagaaaagta gatattgtat ggtggctact aaaaggaaat tcaaaaagtc 
               
               
                   
               
               
                 5461 
                 ttagaacccg agcacctgag caaactgcag tagtcaaaat atttatctca tgttaaagaa 
               
               
                   
               
               
                 5521 
                 aggcaaatct agtgtaagaa atgagtacca tatagggttt tgaagttcat atactagaaa 
               
               
                   
               
               
                 5581 
                 cacttaaaag atatcatttc agatattacg tttggcattg ttcttaagta tttatatctt 
               
               
                   
               
               
                 5641 
                 tgagtcaagc tgataattaa aaaaaatctg ttaatggagt gtatatttca taatgtatca 
               
               
                   
               
               
                 5701 
                 aaatggtgtc tatacctaag gtagcattat tgaagagaga tatgtttatg tagtaagtta 
               
               
                   
               
               
                 5761 
                 ttaacataat gagtaacaaa taatgtttcc agaagaaagg aaaacacatt ttcagagtgc 
               
               
                   
               
               
                 5821 
                 gtttttatca gaggaagaca aaaatacaca cccctctcca gtagcttatt tttacaaagc 
               
               
                   
               
               
                 5881 
                 cggcccagtg aattagaaaa acaaagcact tggatatgat ttttggaaag cccaggtaca 
               
               
                   
               
               
                 5941 
                 cttattattc aaaatgcact tttactgagt ttgaaaagtt tcttttatat ttaaaataag 
               
               
                   
               
               
                 6001 
                 ggttcaaata tgcatattca atttttatag tagttatcta tttgcaaagc atatattaac 
               
               
                   
               
               
                 6061 
                 tagtaattgg ctgttaattt tatagacatg gtagccaggg aagtatatca atgacctatt 
               
               
                   
               
               
                 6121 
                 aagtattttg acaagcaatt tacatatctg atgacctcgt atctcttttt cagcaagtca 
               
               
                   
               
               
                 6181 
                 aatgctatgt aattgttcca ttgtgtgttg tataaaatga atcaacacgg taagaaaaag 
               
               
                   
               
               
                 6241 
                 gttagagtta ttaaaataat aaactgacta aaatactcat ttgaatttat tcagaatgtt 
               
               
                   
               
               
                 6301 
                 cataatgctt tcaaaggaca tagcagagct tttgtggagt atccgcacaa cattatttat 
               
               
                   
               
               
                 6361 
                 tatctatgga ctaaatcaat tttttgaagt tgctttaaaa tttaaaagca cctttgctta 
               
               
                   
               
               
                 6421 
                 atataaagcc ctttaatttt aactgacaga tcaattctga aactttattt tgaaaagaaa 
               
               
                   
               
               
                 6481 
                 atggggaaga atctgtgtct ttagaattaa aagaaatgaa aaaaataaac ccgacattct 
               
               
                   
               
               
                 6541 
                 aaaaaaatag aataagaaac ctgattttta gtactaatga aatagcgggt gacaaaatag 
               
               
                   
               
               
                 6601 
                 ttgtcttttt gattttgatc acaaaaaata aactggtagt gacaggatat gatggagaga 
               
               
                   
               
               
                 6661 
                 tttgacatcc tggcaaatca ctgtcattga ttcaattatt ctaattctga ataaaagctg 
               
               
                   
               
               
                 6721 
                 tatacagtaa aa 
               
            
           
         
       
     
     By “PDX1 polypeptide” is meant a protein or fragment thereof having at least 85% amino acid sequence identity to the sequence provided at NCBI Accession No. NP_000200.1 and having transcription factor activity. The amino acid sequence provided at NCBI Accession No. NP_000200.1 is shown below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 43) 
                   
               
            
           
           
               
               
               
            
               
                   1 
                 mngeeqyyaa tqlykdpcaf qrgpapefsa sppaclymgr qpppppphpf pgalgaleqg 
                   
               
               
                   
               
               
                  61 
                 sppdispyev ppladdpava hlhhhlpaql alphppagpf pegaepgvle epnrvqlpfp 
               
               
                   
               
               
                 121 
                 wmkstkahaw kgqwaggaya aepeenkrtr taytraqlle lekeflfnky isrprrvela 
               
               
                   
               
               
                 181 
                 vmlnlterhi kiwfqnrrmk wkkeedkkrg ggtavggggv aepeqdcavt sgeellalpp 
               
               
                   
               
               
                 241 
                 ppppggavpp aapvaaregr lppglsaspq pssvaprrpq epr 
               
            
           
         
       
     
     By “PDX1 polynucleotide” is meant a polynucleotide encoding a PDX1 polypeptide or fragment thereof. An exemplary PDX1 polynucleotide sequence is provided at NCBI Ref: NM_000209.3. The sequence provided at NCBI Ref: NM_000209.3 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 44) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 gggtggcgcc gggagtggga acgccacaca gtgccaaatc cccggctcca gctcccgact 
                   
               
               
                   
               
               
                 61 
                 cccggctccc ggctcccggc tcccggtgcc caatcccggg ccgcagccat gaacggcgag 
               
               
                   
               
               
                 121 
                 gagcagtact acgcggccac gcagctttac aaggacccat gcgcgttcca gcgaggcccg 
               
               
                   
               
               
                 181 
                 gcgccggagt tcagcgccag cccccctgcg tgcctgtaca tgggccgcca gcccccgccg 
               
               
                   
               
               
                 241 
                 ccgccgccgc acccgttccc tggcgccctg ggcgcgctgg agcagggcag ccccccggac 
               
               
                   
               
               
                 301 
                 atctccccgt acgaggtgcc ccccctcgcc gacgaccccg cggtggcgca ccttcaccac 
               
               
                   
               
               
                 361 
                 cacctcccgg ctcagctcgc gctcccccac ccgcccgccg ggcccttccc ggagggagcc 
               
               
                   
               
               
                 421 
                 gagccgggcg tcctggagga gcccaaccgc gtccagctgc ctttcccatg gatgaagtct 
               
               
                   
               
               
                 481 
                 accaaagctc acgcgtggaa aggccagtgg gcaggcggcg cctacgctgc ggagccggag 
               
               
                   
               
               
                 541 
                 gagaacaagc ggacgcgcac ggcctacacg cgcgcacagc tgctagagct ggagaaggag 
               
               
                   
               
               
                 601 
                 ttcctattca acaagtacat ctcacggccg cgccgggtgg agctggctgt catgttgaac 
               
               
                   
               
               
                 661 
                 ttgaccgaga gacacatcaa gatctggttc caaaaccgcc gcatgaagtg gaaaaaggag 
               
               
                   
               
               
                 721 
                 gaggacaaga agcgcggcgg cgggacagct gtcgggggtg gcggggtcgc ggagcctgag 
               
               
                   
               
               
                 781 
                 caggactgcg ccgtgacctc cggcgaggag cttctggcgc tgccgccgcc gccgcccccc 
               
               
                   
               
               
                 841 
                 ggaggtgctg tgccgcccgc tgcccccgtt gccgcccgag agggccgcct gccgcctggc 
               
               
                   
               
               
                 901 
                 cttagcgcgt cgccacagcc ctccagcgtc gcgcctcggc ggccgcagga accacgatga 
               
               
                   
               
               
                 961 
                 gaggcaggag ctgctcctgg ctgaggggct tcaaccactc gccgaggagg agcagagggc 
               
               
                   
               
               
                 1021 
                 ctaggaggac cccgggcgtg gaccacccgc cctggcagtt gaatggggcg gcaattgcgg 
               
               
                   
               
               
                 1081 
                 ggcccacctt agaccgaagg ggaaaacccg ctctctcagg cgcatgtgcc agttggggcc 
               
               
                   
               
               
                 1141 
                 ccgcgggtag atgccggcag gccttccgga agaaaaagag ccattggttt ttgtagtatt 
               
               
                   
               
               
                 1201 
                 ggggccctct tttagtgata ctggattggc gttgtttgtg gctgttgcgc acatccctgc 
               
               
                   
               
               
                 1261 
                 cctcctacag cactccacct tgggacctgt ttagagaagc cggctcttca aagacaatgg 
               
               
                   
               
               
                 1321 
                 aaactgtacc atacacattg gaaggctccc taacacacac agcggggaag ctgggccgag 
               
               
                   
               
               
                 1381 
                 taccttaatc tgccataaag ccattcttac tcgggcgacc cctttaagtt tagaaataat 
               
               
                   
               
               
                 1441 
                 tgaaaggaaa tgtttgagtt ttcaaagatc ccgtgaaatt gatgccagtg gaatacagtg 
               
               
                   
               
               
                 1501 
                 agtcctcctc ttcctcctcc tcctcttccc cctccccttc ctcctcctcc tcttcttttc 
               
               
                   
               
               
                 1561 
                 cctcctcttc ctcttcctcc tgctctcctt tcctccccct cctcttttcc ctcctcttcc 
               
               
                   
               
               
                 1621 
                 tcttcctcct gctctccttt cctccccctc ctctttctcc tcctcctcct cttcttcccc 
               
               
                   
               
               
                 1681 
                 ctcctctccc tcctcctctt cttccccctc ctctccctcc tcctcttctt ctccctcctc 
               
               
                   
               
               
                 1741 
                 ttcctcttcc tcctcttcca cgtgctctcc tttcctcccc ctcctcttgc tccccttctt 
               
               
                   
               
               
                 1801 
                 ccccgtcctc ttcctcctcc tcctcttctt ctccctcctc ttcctcctcc tctttcttcc 
               
               
                   
               
               
                 1861 
                 tgacctcttt ctttctcctc ctcctccttc tacctcccct tctcatccct cctcttcctc 
               
               
                   
               
               
                 1921 
                 ttctctagct gcacacttca ctactgcaca tcttataact tgcacccctt tcttctgagg 
               
               
                   
               
               
                 1981 
                 aagagaacat cttgcaaggc agggcgagca gcggcagggc tggcttagga gcagtgcaag 
               
               
                   
               
               
                 2041 
                 agtccctgtg ctccagttcc acactgctgg cagggaaggc aaggggggac gggcctggat 
               
               
                   
               
               
                 2101 
                 ctgggggtga gggagaaaga tggacccctg ggtgaccact aaaccaaaga tattcggaac 
               
               
                   
               
               
                 2161 
                 tttctattta ggatgtggac gtaattcctg ttccgaggta gaggctgtgc tgaagacaag 
               
               
                   
               
               
                 2221 
                 cacagtggcc tggtgcgcct tggaaaccaa caactattca cgagccagta tgaccttcac 
               
               
                   
               
               
                 2281 
                 atctttagaa attatgaaaa cgtatgtgat tggagggttt ggaaaaccag ttatcttatt 
               
               
                   
               
               
                 2341 
                 taacatttta aaaattacct aacagttatt tacaaacagg tctgtgcatc ccaggtctgt 
               
               
                   
               
               
                 2401 
                 cttcttttca aggtctgggc cttgtgctcg ggttatgttt gtgggaaatg cttaataaat 
               
               
                   
               
               
                 2461 
                 actgataata tgggaagaga tgaaaactga ttctcctcac tttgtttcaa acctttctgg 
               
               
                   
               
               
                 2521 
                 cagtgggatg attcgaattc acttttaaaa ttaaattagc gtgttttgtt ttg 
               
            
           
         
       
     
     By “PTF1 polypeptide” is meant a protein or fragment thereof having at least 85% amino acid sequence identity to the sequence provided at NCBI Accession No. NP_835455.1 and having transcription factor activity. The amino acid sequence provided at NCBI Accession No. NP_835455.1 is shown below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 45) 
                   
               
            
           
           
               
               
               
            
               
                   1 
                 mdavllehfp ggldafpssy fdeddfftdq ssrdpledgd elladeqaev eflshqlhey 
                   
               
               
                   
               
               
                  61 
                 cyrdgaclll qpappaapla lappssgglg epddgggggy ccetgappgg fpyspgspps 
               
               
                   
               
               
                 121 
                 claypcagaa vlspgarlrg lsgaaaaaar rrrrvrseae lqqlrqaanv rerrrmqsin 
               
               
                   
               
               
                 181 
                 dafeglrshi ptlpyekrls kvdtlrlaig yinflselvq adlplrggga ggcggpgggg 
               
               
                   
               
               
                 241 
                 rlggdspgsq aqkviichrg trspspsdpd yglpplaghs lswtdekqlk eqniirtakv 
               
               
                   
               
               
                 301 
                 wtpedprkln skssfnnien eppfefvs 
               
            
           
         
       
     
     By “PTF1 polynucleotide” is meant a polynucleotide encoding a PTF1 polypeptide or fragment thereof. An exemplary PTF1 polynucleotide sequence is provided at NCBI Ref: NM_178161.2. The sequence provided at NCBI Ref: NM_178161.2 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 46) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 atggacgcgg tgttgctgga gcacttcccc gggggcctag acgcctttcc ttcttcgtac 
                   
               
               
                   
               
               
                 61 
                 ttcgacgagg acgacttctt caccgaccag tcttcacggg accccctgga ggacggcgat 
               
               
                   
               
               
                 121 
                 gagctgctgg cggacgagca ggccgaggtg gagttcctta gccaccagct ccacgagtac 
               
               
                   
               
               
                 181 
                 tgctaccgcg acggggcgtg cctgctgctg cagcccgcgc ccccggccgc cccgctagcg 
               
               
                   
               
               
                 241 
                 ctcgccccgc cgtcctcggg gggcctcggt gagccagacg acggcggcgg cggcggctac 
               
               
                   
               
               
                 301 
                 tgctgcgaga cgggggcgcc cccaggcggc ttcccctact cgcccggctc gccgccctcg 
               
               
                   
               
               
                 361 
                 tgcctggcct acccgtgcgc cggggcggca gtactgtctc ccggggcgcg gctgcgcggc 
               
               
                   
               
               
                 421 
                 ctgagcggag cggcggctgc ggcggcgcgg cgccggcggc gggtgcgctc cgaggcggag 
               
               
                   
               
               
                 481 
                 ctgcagcagc tgcggcaggc ggccaacgtg cgcgagcggc ggcgcatgca gtccatcaac 
               
               
                   
               
               
                 541 
                 gacgccttcg aggggctgcg ctcgcacatc cccacgctgc cctacgagaa gcgcctctcc 
               
               
                   
               
               
                 601 
                 aaggtggaca cgctgcgcct ggccatcggc tacatcaact tcctcagcga gctcgtgcag 
               
               
                   
               
               
                 661 
                 gccgacctgc ccttgcgcgg cggtggcgcg ggcggctgcg gggggccggg cggcggcggg 
               
               
                   
               
               
                 721 
                 cgcctgggcg gggacagccc gggcagccag gcccagaagg tcatcatctg ccatcggggc 
               
               
                   
               
               
                 781 
                 acccggtccc cctcccccag cgaccctgat tatggcctcc ctcccctagc aggacactct 
               
               
                   
               
               
                 841 
                 ctctcatgga ctgatgaaaa acaactcaag gaacaaaata ttatccgaac agccaaagtc 
               
               
                   
               
               
                 901 
                 tggaccccag aggaccccag aaaactcaac agcaaatctt ccttcaacaa catagaaaac 
               
               
                   
               
               
                 961 
                 gaaccaccat ttgagtttgt gtcctgagaa gtcccagact cggctgaaga tctgattatg 
               
               
                   
               
               
                 1021 
                 tctctgtgca tattgtacat gtaaatatct ataatgtaaa tgtaatttaa gaatcaaatt 
               
               
                   
               
               
                 1081 
                 tttcgaatgg caatcaactg tttattattt atctatttat tatcctgttg agttgatgaa 
               
               
                   
               
               
                 1141 
                 atagatgatt tctttttaaa tatataattt atataactta tcctgatttt ctgaaaatat 
               
               
                   
               
               
                 1201 
                 gcaatagcct atgattttcc tgaactctgt gttgttggga gaactctggc cagaaaacgt 
               
               
                   
               
               
                 1261 
                 cctgcttatt tattgccaga tatggtttat ttctaagcgt tgtcaataaa tgctatttac 
               
               
                   
               
               
                 1321 
                 accttttcct gaaaaaaaa 
               
            
           
         
       
     
     By “Wnt3a polynucleotide” is meant a polynucleotide encoding a Wnt3a polypeptide or a fragment thereof, or a polynucleotide having at least 85% sequence identity to the human Wnt3a polynucleotide sequence. An exemplary human Wnt3a polynucleotide sequence is provided at NCBI GenBank Accession No. AB060284.1. The polynucleotide sequence provided at NCBI GenBank Accession No. AB060284.1 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 47) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 cggcgatggc cccactcgga tacttcttac tcctctgcag cctgaagcag gctctgggca 
                   
               
               
                   
               
               
                 61 
                 gctacccgat ctggtggtcg ctggctgttg ggccacagta ttcctccctg ggctcgcagc 
               
               
                   
               
               
                 121 
                 ccatcctgtg tgccagcatc ccgggcctgg tccccaagca gctccgcttc tgcaggaact 
               
               
                   
               
               
                 181 
                 acgtggagat catgcccagc gtggccgagg gcatcaagat tggcatccag gagtgccagc 
               
               
                   
               
               
                 241 
                 accagttccg cggccgccgg tggaactgca ccaccgtcca cgacagcctg gccatcttcg 
               
               
                   
               
               
                 301 
                 ggcccgtgct ggacaaagct accagggagt cggcctttgt ccacgccatt gcctcagccg 
               
               
                   
               
               
                 361 
                 gtgtggcctt tgcagtgaca cgctcatgtg cagaaggcac ggccgccatc tgtggctgca 
               
               
                   
               
               
                 421 
                 gcagccgcca ccagggctca ccaggcaagg gctggaagtg gggtggctgt agcgaggaca 
               
               
                   
               
               
                 481 
                 tcgagtttgg tgggatggtg tctcgggagt tcgccgacgc ccgggagaac cggccagatg 
               
               
                   
               
               
                 541 
                 cccgctcagc catgaaccgc cacaacaacg aggctgggcg ccaggccatc gccagccaca 
               
               
                   
               
               
                 601 
                 tgcacctcaa gtgcaagtgc cacgggctgt cgggcagctg cgaggtgaag acatgctggt 
               
               
                   
               
               
                 661 
                 ggtcgcaacc cgacttccgc gccatcggtg acttcctcaa ggacaagtac gacagcgcct 
               
               
                   
               
               
                 721 
                 cggagatggt ggtggagaag caccgggagt cccgcggctg ggtggagacc ctgcggccgc 
               
               
                   
               
               
                 781 
                 gctacaccta cttcaaggtg cccacggagc gcgacctggt ctactacgag gcctcgccca 
               
               
                   
               
               
                 841 
                 acttctgcga gcccaaccct gagacgggct ccttcggcac gcgcgaccgc acctgcaacg 
               
               
                   
               
               
                 901 
                 tcagctcgca cggcatcgac ggctgcgacc tgctgtgctg cggccgcggc cacaacgcgc 
               
               
                   
               
               
                 961 
                 gagcggagcg gcgccgggag aagtgccgct gcgtgttcca ctggtgctgc tacgtcagct 
               
               
                   
               
               
                 1021 
                 gccaggagtg cacgcgcgtc tacgacgtgc acacctgcaa gtaggcaccg gccgcggctc 
               
               
                   
               
               
                 1081 
                 cccctggacg gggcgggccc tgcctgaggg tgggcttttc cctgggtgga gcaggactcc 
               
               
                   
               
               
                 1141 
                 cacctaaacg gggcagtact cctccctggg ggcgggactc ctccctgggg gtggggctcc 
               
               
                   
               
               
                 1201 
                 tacctggggg cagaactcct acctgaaggc agggctcctc cctggagcta gtgtctcctc 
               
               
                   
               
               
                 1261 
                 tctggtggct gggctgctcc tgaatgaggc ggagctccag gatggggagg ggctctgcgt 
               
               
                   
               
               
                 1321 
                 tggcttctcc ctggggacgg ggctcccctg gacagaggcg gggctacaga ttgggcgggg 
               
               
                   
               
               
                 1381 
                 cttctcttgg gtgggacagg gcttctcctg cgggggcgag gcccctccca gtaagggcgt 
               
               
                   
               
               
                 1441 
                 ggctctgggt gggcggggca ctaggtaggc ttctacctgc aggcggggct cctcctgaag 
               
               
                   
               
               
                 1501 
                 gaggcggggc tctaggatgg ggcacggctc tggggtaggc tgctccctga gggcg 
               
            
           
         
       
     
     By “Wnt3a polypeptide” is meant a Wnt3a polypeptide or a fragment thereof, or a polypeptide having at least 85% sequence identity to the human Wnt3a polypeptide sequence. An exemplary human Wnt3a polypeptide sequence is provided at NCBI GenBank: AAI03924.1. The sequence provided at GenBank: AAI03924.1 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 48) 
                   
               
            
           
           
               
               
               
            
               
                   1 
                 maplgyflll cslkqalgsy piwwslavgp qysslgsqpi lcasipglvp kqlrfcrnyv 
                   
               
               
                   
               
               
                  61 
                 eimpsvaegi kigiqecqhq frgrrwnctt vhdslaifgp vldkatresa fvhaiasagv 
               
               
                   
               
               
                 121 
                 afavtrscae gtaaicgcss rhqgspgkgw kwggcsedie fggmvsrefa darenrpdar 
               
               
                   
               
               
                 181 
                 samnrhnnea grqaiashmh lkckchglsg scevktcwws qpdfraigdf lkdkydsase 
               
               
                   
               
               
                 241 
                 mvvekhresr gwvetlrpry tyfkvpterd lvyyeaspnf cepnpetgsf gtrdrtcnvs 
               
               
                   
               
               
                 301 
                 shgidgcdll ccgrghnara errrekcrcv fhwccyvscq ectrvydvht cknpgsragn 
               
               
                   
               
               
                 361 
                 sahqpphpqp pvrfhpplrr agkvp 
               
            
           
         
       
     
     By “Wnt4 polynucleotide” is meant a polynucleotide encoding Wnt4 polypeptide or a fragment thereof, or a polynucleotide having at least 85% sequence identity to the human Wnt4 polynucleotide sequence. An exemplary human Wnt4 polynucleotide sequence is provided at NCBI GenBank Accession No. AY009398.1. Accession number NCBI Ref NG 008974.1 is a reference standard Wnt4a polynucleotide sequence. The polynucleotide sequence provided at NCBI GenBank Accession No. AY009398.1 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 49) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 atgagtcccc gctcgtgcct gcgttcgctg cgcctcctcg tcttcgccgt cttctcagcc 
                   
               
               
                   
               
               
                 61 
                 gccgcgagca actggctgta cctggccaag ctgtcgtcgg tggggagcat ctcagaggag 
               
               
                   
               
               
                 121 
                 gagacgtgcg agaaactcaa gggcctgatc cagaggcagg tgcagatgtg caagcggaac 
               
               
                   
               
               
                 181 
                 ctggaagtca tggactcggt gcgccgcggt gcccagctgg ccattgagga gtgccagtac 
               
               
                   
               
               
                 241 
                 cagttccgga accggcgctg gaactgctcc acactcgact ccttgcccgt cttcggcaag 
               
               
                   
               
               
                 301 
                 gtggtgacgc aagggattcg ggaggcggcc ttggtgtacg ccatctcttc ggcaggtgtg 
               
               
                   
               
               
                 361 
                 gcctttgcag tgacgcgggc gtgcagcagt ggggagctgg agaagtgcgg ctgtgacagg 
               
               
                   
               
               
                 421 
                 acagtgcatg gggtcagccc acagggcttc cagtggtcag gatgctctga caacatcgcc 
               
               
                   
               
               
                 481 
                 tacggtgtgg ccttctcaca gtcgtttgtg gatgtgcggg agagaagcaa gggggcctcg 
               
               
                   
               
               
                 541 
                 tccagcagag ccctcatgaa cctccacaac aatgaggccg gcaggaaggc catcctgaca 
               
               
                   
               
               
                 601 
                 cacatgcggg tggaatgcaa gtgccacggg gtgtcaggct cctgtgaggt aaagacgtgc 
               
               
                   
               
               
                 661 
                 tggcgagccg tgccgccctt ccgccaggtg ggtcacgcac tgaaggagaa gtttgatggt 
               
               
                   
               
               
                 721 
                 gccactgagg tggagccacg ccgcgtgggc tcctccaggg cactggtgcc acgcaacgca 
               
               
                   
               
               
                 781 
                 cagttcaagc cgcacacaga tgaggacttg gtgtacttgg agcctagccc cgacttctgt 
               
               
                   
               
               
                 841 
                 gagcaggaca tgcgcagcgg cgtgctgggc acgaggggcc gcacatgcaa caagacgtcc 
               
               
                   
               
               
                 901 
                 aaggccatcg acggctgtga gctgctgtgc tgtggccgcg gcttccacac ggcgcaggtg 
               
               
                   
               
               
                 961 
                 gagctggctg aacgctgcag ctgcaaattc cactggtgct gcttcgtcaa gtgccggcag 
               
               
                   
               
               
                 1021 
                 tgccagcggc tcgtggagtt gcacacgtgc cgatga 
               
            
           
         
       
     
     By “Wnt4 polypeptide” is meant a Wnt4 polypeptide or a fragment thereof, or a polypeptide having at least 85% sequence identity to the human Wnt4 polypeptide sequence. An exemplary human Wnt4 polypeptide sequence is provided at NCBI GenBank Accession No.: AAG38658.1. The sequence provided at GenBank Accession No.: AAG38658.1 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 50) 
                   
               
            
           
           
               
               
               
            
               
                   1 
                 msprsclrsl rllvfavfsa aasnwlylak lssvgsisee etceklkgli qrqvqmckrn 
                   
               
               
                   
               
               
                  61 
                 levmdsvrrg aqlaieecqy qfrnrrwncs tldslpvfgk vvtqgireaa lvyaissagv 
               
               
                   
               
               
                 121 
                 afavtracss gelekcgcdr tvhgvspqgf qwsgcsdnia ygvafsqsfv dvrerskgas 
               
               
                   
               
               
                 181 
                 ssralmnlhn neagrkailt hmrveckchg vsgscevktc wravppfrqv ghalkekfdg 
               
               
                   
               
               
                 241 
                 ateveprrvg ssralvprna qfkphtdedl vylepspdfc eqdmrsgvlg trgrtcnkts 
               
               
                   
               
               
                 301 
                 kaidgcellc cgrgfhtaqv elaercsckf hwccfvkcrq cqrlvelhtc r 
               
            
           
         
       
     
     By “Wnt5a polynucleotide” is meant a polynucleotide encoding Wnt5a polypeptide or a fragment thereof, or a polynucleotide having at least 85% sequence identity to the human Wnt5a polynucleotide sequence. An exemplary polynucleotide sequence coding for human Wnt5a is provided at NCBI Ref: GenBank NM_003392, a reference standard sequence. Nucleotides 658-1800 of the Wnt5a genomic sequence having 6194 nucleotides codes for a human Wnt5a polypeptide. The polynucleotide sequence of the human Wnt5a coding sequence provided at bases 658-1800 of NCBI Ref: GenBank NM_003392 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 51) 
                   
               
            
           
           
               
               
               
            
               
                  658 
                 atg 
                   
               
               
                   
               
               
                  661 
                 aagaagtcca ttggaatatt aagcccagga gttgctttgg ggatggctgg aagtgcaatg 
               
               
                   
               
               
                  721 
                 tcttccaagt tcttcctagt ggctttggcc atatttttct ccttcgccca ggttgtaatt 
               
               
                   
               
               
                  781 
                 gaagccaatt cttggtggtc gctaggtatg aataaccctg ttcagatgtc agaagtatat 
               
               
                   
               
               
                  841 
                 attataggag cacagcctct ctgcagccaa ctggcaggac tttctcaagg acagaagaaa 
               
               
                   
               
               
                  901 
                 ctgtgccact tgtatcagga ccacatgcag tacatcggag aaggcgcgaa gacaggcatc 
               
               
                   
               
               
                  961 
                 aaagaatgcc agtatcaatt ccgacatcga aggtggaact gcagcactgt ggataacacc 
               
               
                   
               
               
                 1021 
                 tctgtttttg gcagggtgat gcagataggc agccgcgaga cggccttcac atacgcggtg 
               
               
                   
               
               
                 1081 
                 agcgcagcag gggtggtgaa cgccatgagc cgggcgtgcc gcgagggcga gctgtccacc 
               
               
                   
               
               
                 1141 
                 tgcggctgca gccgcgccgc gcgccccaag gacctgccgc gggactggct ctggggcggc 
               
               
                   
               
               
                 1201 
                 tgcggcgaca acatcgacta tggctaccgc tttgccaagg agttcgtgga cgcccgcgag 
               
               
                   
               
               
                 1261 
                 cgggagcgca tccacgccaa gggctcctac gagagtgctc gcatcctcat gaacctgcac 
               
               
                   
               
               
                 1321 
                 aacaacgagg ccggccgcag gacggtgtac aacctggctg atgtggcctg caagtgccat 
               
               
                   
               
               
                 1381 
                 ggggtgtccg gctcatgtag cctgaagaca tgctggctgc agctggcaga cttccgcaag 
               
               
                   
               
               
                 1441 
                 gtgggtgatg ccctgaagga gaagtacgac agcgcggcgg ccatgcggct caacagccgg 
               
               
                   
               
               
                 1501 
                 ggcaagttgg tacaggtcaa cagccgcttc aactcgccca ccacacaaga cctggtctac 
               
               
                   
               
               
                 1561 
                 atcgacccca gccctgacta ctgcgtgcgc aatgagagca ccggctcgct gggcacgcag 
               
               
                   
               
               
                 1621 
                 ggccgcctgt gcaacaagac gtcggagggc atggatggct gcgagctcat gtgctgcggc 
               
               
                   
               
               
                 1681 
                 cgtggctacg accagttcaa gaccgtgcag acggagcgct gccactgcaa gttccactgg 
               
               
                   
               
               
                 1741 
                 tgctgctacg tcaagtgcaa gaagtgcacg gagatcgtgg accagtttgt gtgcaagtag 
               
            
           
         
       
     
     By “Wnt5a polypeptide” is meant a Wnt5a polypeptide or a fragment thereof, or a polypeptide having at least 85% sequence identity to the human Wnt5a polypeptide sequence. An exemplary human Wnt5a (isoform 1) polypeptide sequence is provided at UniProtKB Identifier: P41221-1. The sequence provided at UniProtKB Identifier: P41221-1 is reproduced below: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 52) 
                   
               
            
           
           
               
               
               
            
               
                   1 
                 mkksigilsp gvalgmagsa msskfflval aiffsfaqvv ieanswwslg 
                   
               
               
                   
               
               
                  51 
                 mnnpvqmsev yiigaqplcs qlaglsqgqk klchlyqdhm qyigegaktg 
               
               
                   
               
               
                 101 
                 ikecqyqfrh rrwncstvdn tsvfgrvmqi gsretaftya vsaagvvnam 
               
               
                   
               
               
                 151 
                 sracregels tcgcsraarp kdlprdwlwg gcgdnidygy rfakefvdar 
               
               
                   
               
               
                 201 
                 ererihakgs yesarilmnl hnneagrrtv ynladvackc hgvsgscslk 
               
               
                   
               
               
                 251 
                 tcwlqladfr kvgdalkeky dsaaamrlns rgklvqvnsr fnspttqdlv 
               
               
                   
               
               
                 301 
                 yidpspdycv rnestgslgt qgrlcnktse gmdgcelmcc grgydqfktv 
               
               
                   
               
               
                 351 
                 qterchckfh wccyvkckkc teivdqfvck 
               
            
           
         
       
     
     By “progenitor cell” is meant a cell that a multipotent stem cell that is capable of generating (e.g., by differentiation or division) an endothelial cell. A progenitor cell that is capable of generating an endothelial cell may express this capability when grown under appropriate in vitro or in vivo conditions, such as those described herein. 
     By “progeny” is meant a cell derived from a multipotent stem cell of the invention. Progeny include without limitation progenitor cells, differentiated cells, and terminally differentiated cells. 
     By “derived from” is meant the process of obtaining a progeny cell. 
     By “reduces” is meant a negative alteration of at least 10%, 25%, 50%, 75%, or 100%. 
     By “reference” or “control” is meant a standard condition. For example, an untreated cell, tissue, or organ that is used as a reference. 
     A “reference sequence” is a defined sequence used as a basis for sequence comparison. A reference sequence may be a subset of or the entirety of a specified sequence; for example, a segment of a full-length cDNA or gene sequence, or the complete cDNA or gene sequence. For polypeptides, the length of the reference polypeptide sequence will generally be at least about 16 amino acids, at least about 20 amino acids, or at least about 25 amino acids. The length of the reference polypeptide sequence can be about 35 amino acids, about 50 amino acids, or about 100 amino acids. For nucleic acids, the length of the reference nucleic acid sequence will generally be at least about 50 nucleotides, at least about 60 nucleotides, or at least about 75 nucleotides. The length of the reference nucleic acid sequence can be about 100 nucleotides, about 300 nucleotides or any integer thereabout or therebetween. 
     A “somatic” cell refers to a cell that is obtained from a tissue of a subject. Such subjects are at a post-natal stage of development (e.g., adult, infant, child). In contrast, an “embryonic cell” or “embryonic stem cell” is derived from an embryo at a pre-natal stage of development. 
     By “specifically binds” is meant a compound or antibody that recognizes and binds a polypeptide of the invention, but which does not substantially recognize and bind other molecules in a sample, for example, a biological sample, which naturally includes a polypeptide of the invention. 
     Nucleic acid molecules useful in the methods of the invention include any nucleic acid molecule that encodes a polypeptide of the invention or a fragment thereof. Such nucleic acid molecules need not be 100% identical with an endogenous nucleic acid sequence, but will typically exhibit substantial identity. Polynucleotides having “substantial identity” to an endogenous sequence are typically capable of hybridizing with at least one strand of a double-stranded nucleic acid molecule. Nucleic acid molecules useful in the methods of the invention include any nucleic acid molecule that encodes a polypeptide of the invention or a fragment thereof. Such nucleic acid molecules need not be 100% identical with an endogenous nucleic acid sequence, but will typically exhibit substantial identity. Polynucleotides having “substantial identity” to an endogenous sequence are typically capable of hybridizing with at least one strand of a double-stranded nucleic acid molecule. By “hybridize” is meant pair to form a double-stranded molecule between complementary polynucleotide sequences (e.g., a gene described herein), or portions thereof, under various conditions of stringency. (See, e.g., Wahl, G. M. and S. L. Berger (1987) Methods Enzymol. 152:399; Kimmel, A. R. (1987) Methods Enzymol. 152:507). 
     For example, stringent salt concentration will ordinarily be less than about 750 mM NaCl and 75 mM trisodium citrate, less than about 500 mM NaCl and 50 mM trisodium citrate, or less than about 250 mM NaCl and 25 mM trisodium citrate. Low stringency hybridization can be obtained in the absence of organic solvent, e.g., formamide, while high stringency hybridization can be obtained in the presence of at least about 35% formamide, or at least about 50% formamide. Stringent temperature conditions will ordinarily include temperatures of at least about 30° C., at least about 37° C., and at least about 42° C. Varying additional parameters, such as hybridization time, the concentration of detergent, e.g., sodium dodecyl sulfate (SDS), and the inclusion or exclusion of carrier DNA, are well known to those skilled in the art. Various levels of stringency are accomplished by combining these various conditions as needed. In one embodiment, hybridization will occur at 30° C. in 750 mM NaCl, 75 mM trisodium citrate, and 1% SDS. In another embodiment, hybridization will occur at 37° C. in 500 mM NaCl, 50 mM trisodium citrate, 1% SDS, 35% formamide, and 100 μg/ml denatured salmon sperm DNA (ssDNA). In yet another embodiment, hybridization will occur at 42° C. in 250 mM NaCl, 25 mM trisodium citrate, 1% SDS, 50% formamide, and 200 μg/ml ssDNA. Useful variations on these conditions will be readily apparent to those skilled in the art. 
     For most applications, washing steps that follow hybridization will also vary in stringency. Wash stringency conditions can be defined by salt concentration and by temperature. As above, wash stringency can be increased by decreasing salt concentration or by increasing temperature. For example, stringent salt concentration for the wash steps will be less than about 30 mM NaCl and 3 mM trisodium citrate, or less than about 15 mM NaCl and 1.5 mM trisodium citrate. Stringent temperature conditions for the wash steps will ordinarily include a temperature of at least about 25° C., at least about 42° C., and at least about 68° C. In one embodiment, wash steps will occur at 25° C. in 30 mM NaCl, 3 mM trisodium citrate, and 0.1% SDS. In another embodiment, wash steps will occur at 42 C in 15 mM NaCl, 1.5 mM trisodium citrate, and 0.1% SDS. In yet another embodiment, wash steps will occur at 68° C. in 15 mM NaCl, 1.5 mM trisodium citrate, and 0.1% SDS. Additional variations on these conditions will be readily apparent to those skilled in the art. Hybridization techniques are well known to those skilled in the art and are described, for example, in Benton and Davis (Science 196:180, 1977); Grunstein and Hogness (Proc. Natl. Acad. Sci., USA 72:3961, 1975); Ausubel et al. (Current Protocols in Molecular Biology, Wiley Interscience, New York, 2001); Berger and Kimmel (Guide to Molecular Cloning Techniques, 1987, Academic Press, New York); and Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, New York. 
     By “substantially identical” is meant a polypeptide or nucleic acid molecule exhibiting at least 50% identity to a reference amino acid sequence (for example, any one of the amino acid sequences described herein) or nucleic acid sequence (for example, any one of the nucleic acid sequences described herein). Such a sequence is at least 60%, at least 80%, at least 85%, at least 90%, at least 95% or even at least 99% identical at the amino acid level or nucleic acid to the sequence used for comparison. 
     Sequence identity is typically measured using sequence analysis software (for example, Sequence Analysis Software Package of the Genetics Computer Group, University of Wisconsin Biotechnology Center, 1710 University Avenue, Madison, Wis. 53705, BLAST, BESTFIT, GAP, or PILEUP/PRETTYBOX programs). Such software matches identical or similar sequences by assigning degrees of homology to various substitutions, deletions, and/or other modifications. Conservative substitutions typically include substitutions within the following groups: glycine, alanine; valine, isoleucine, leucine; aspartic acid, glutamic acid, asparagine, glutamine; serine, threonine; lysine, arginine; and phenylalanine, tyrosine. In an exemplary approach to determining the degree of identity, a BLAST program may be used, with a probability score between e −3  and e −100  indicating a closely related sequence. 
     The term “self renewal” as used herein refers to the process by which a stem cell divides to generate one (asymmetric division) or two (symmetric division) daughter cells with development potentials that are indistinguishable from those of the mother cell. Self renewal involves both proliferation and the maintenance of an undifferentiated state. 
     The term “stem cell” is meant a pluripotent cell or multipotent stem cell having the capacity to self-renew and to differentiate into multiple cell lineages. 
     By “subject” is meant a mammal, including, but not limited to, a human or non-human mammal, such as a bovine, equine, canine, ovine, rodent, or feline. 
     Ranges provided herein are understood to be shorthand for all of the values within the range. For example, a range of 1 to 50 is understood to include any number, combination of numbers, or sub-range from the group consisting 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50. 
     By “tissue” is meant a collection of cells having a similar morphology and function. As used herein, the terms “treat,” treating,” “treatment,” and the like refer to reducing or ameliorating a disorder and/or symptoms associated therewith. It will be appreciated that, although not precluded, treating a disorder or condition does not require that the disorder, condition or symptoms associated therewith be completely eliminated. 
     By “vascularized” is meant having a blood vessel. In some embodiments, the pancreatic islet organoid or pancreatic organoid is vascularized. 
     Unless specifically stated or obvious from context, as used herein, the term “or” is understood to be inclusive. Unless specifically stated or obvious from context, as used herein, the terms “a”, “an”, and “the” are understood to be singular or plural. 
     Unless specifically stated or obvious from context, as used herein, the term “about” is understood as within a range of normal tolerance in the art, for example within 2 standard deviations of the mean. About can be understood as within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the stated value. Unless otherwise clear from context, all numerical values provided herein are modified by the term about. 
     The recitation of a listing of chemical groups in any definition of a variable herein includes definitions of that variable as any single group or combination of listed groups. The recitation of an embodiment for a variable or aspect herein includes that embodiment as any single embodiment or in combination with any other embodiments or portions thereof. 
     Any compositions or methods provided herein can be combined with one or more of any of the other compositions and methods provided herein. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG. 1  is a schematic showing the various symptoms of human disease that can be modeled in a dish. Generation of functional human organs provides new therapeutic strategies in drug-screening and disease modeling. Described herein is a novel technique to generate 3D human mini-organs in a dish. Using this technique, human type 2 diabetes can be modeled in a dish to find effective drugs in genetic, patients or environmental specific diseases such as human type 2 diabetes. 
         FIG. 2A  is a set of micrographs showing the utility of human adipose-derived stem cells (hADSC) in organogenesis. In studies described herein, human adipose-derived stem cells were found to be a novel resource for generation of self-organized organoids (organ bud).  FIG. 2  compares the appearance of human iPSC-derived pancreatic progenitors (PPs. Day 15-day 19), human umbilical vein endothelial cells (HUVEC), and hADSCs cultured on plates (2D) with those cultured in the 3 dimensional matrigel (Matrigel® system for 24 hours in PP cell differentiation media, endothelial cell growth media, and hADSC growth media, respectively.  FIG. 2B  is a set of micrographs demonstrating the ability of ADSCs to progressively self-organize when cultured in Matrigel.  FIG. 2C  is a set of images of hADSCs, seeded at the indicated density, that demonstrate the minimum number of cells required for sphere formation when grown in Matrigel.  FIGS. 2D and 2D-1 to 2D-4  show transcriptional changes occurring in hADSCs during 48 hours of culture in Matrigel, depicted as a heatmap. Statistically different changes in gene expression were determined by RNA-Seq analyses of cells after the indicated time in Matrigel. Biological pathways altered during culture in Matrigel were identified using DAVID software.  FIG. 2E  is a schematic describing the generation of human islet-like organoids by culturing in Matrigel.  FIG. 2F  is an image of human islet-like organoids in a single well of a 24 well plate.  FIG. 2G  is a set of images of a human islet-like organoid generated by co-culturing hiPSC-derived pancreatic progenitors, HUVECs, and hADSCs for 1-5 days in Matrigel. GFP is used to indicate human insulin expression (green, 1 st  panel), mCherry to label HUVEC cells (red, 2 nd  panel), brightfield image (3 rd  panel), and an overlay of GFP and mCherry images (4 th  panel). 
         FIGS. 3A-3F  are plots, images, and a schematic showing characterization of insulin secretion of INS-1 cells, mouse islets, and human islets using a proinsulin luciferase reporter system as a quantitative insulin secretion assay.  FIG. 3A  is a schematic showing the generation of a stably-expressed insulin reporter in the rat beta cell line INS-1 cells, where luciferase expression is under the control of the proinsulin promoter.  FIG. 3B  is a plot showing luciferase activity induced in the INS-1 reporter cells in response to treatment with 3 mM glucose (G3), 20 mM glucose (G20), 20 mM glucose (G20) plus 100 nM Exendin-4 (Ex-4), or 20 mM potassium chloride (KCl).  FIG. 3C  is an image showing a single mouse islet in one well of a 96-well plate. The mouse islet cells were infected with the proinsulin-luciferase lentivirus reporter construct 2 days before assaying.  FIG. 3D  is a set of plots showing the luciferase activity from individual mouse islets infected with the proinsulin luciferase reporter in response to 3 mM glucose (G3 mM) and 20 mM glucose (G20 mM) (left), and the average of the individual assays (right).  FIG. 3E  is an image showing a single human islet in one well of a 96-well plate. The human islet cells were infected with the proinsulin-luciferase lentivirus reporter construct 2 days before assaying.  FIG. 3F  is a set of plots showing the luciferase activity from individual human islets infected with the proinsulin luciferase reporter in response to 3 mM glucose (G3 mM) and 20 mM glucose (G20 mM) (left), and the average of the individual assays (right).  FIGS. 3D and 3F  show that glucose-stimulated insulin secretion (GSIS) can be measured from single mouse and human islets, respectively, after the infection with the lentiviral luciferase reporter. 
         FIGS. 4A-4I  are plots, images, graphs and a schematic showing generation of functional, vascularized human pancreatic islets in a dish.  FIG. 4A  is a schematic showing a scheme for the generation of functional, vascularized human pancreatic islets in Gellan gum. 
         FIG. 4B  shows human islet-like mini organs. Top panels show insulin positive cells (green fluorescent protein expression driven by the insulin promoter (left)) and phase contrast images (left) of islet-like organoids grown in 3D Gellan gum suspensions (bottom panel). Electron microscopy images reveal insulin granules in the β-like cells and lipid droplets in the hADSCs.  FIG. 4C  shows that human islet-like-mini organs generated by methods described herein are morphologically identical to human islets.  FIG. 4D  and  FIG. 4E  show relative expression of genes associated with β cell determination ( FIG. 4D ), and mitochondrial function ( FIG. 4E ), as measured by qPCR. Islet-like cell clusters (derived from pancreatic progenitors cultured in gellan gum, day 35) and islet-like organoids (derived from pancreatic progenitors co-cultured with HUVECs and hADSCs in gellan gum) were FACS stored into insulin expressing (GFP positive) and non-expressing (GFP negative) cells prior to analysis, and compared with gene expression in human islets.  FIG. 4F  shows glucose-stimulated insulin secretion, measured by the fold change in c-peptide secretion 30 minutes after exposure to 20 mM glucose, in selected islet-like organoid preparations prepared in 3D gellan gum cultures and human islets.  FIG. 4G  shows that human islet-like mini organs generated by the methods described herein can develop functional vascularization. Human islet-like organoids were transferred to matrigel and grown in the presence of endothelial growth media. Cells expressing insulin are visualized as green fluorescence. Top panels are fluorescent images of cells showing HUVEC cell outgrowth 24 and 48 hours after stimulation by endothelial cell growth medium (ECM). The bottom panel is a schematic summarizing the experiment and the finding.  FIG. 4H  shows a graph illustrating that human islet-like mini organs generated by the methods described herein can regulate blood glucose in a known mouse model of type 1 diabetes, NODSCID. In this mouse model, the SCID (Severe Combined Immune Deficiency) mutation has been transferred onto a diabetes-susceptible Non-Obese Diabetic (NOD) background. The multiple immunological defects in this mouse model provide a system for reconstituting the animal with human hematopoietic cells. The graph in  FIG. 4H  shows the blood glucose levels in NODSCID mice treated with streptozotocin (STZ) (180 mg/kg) to induce type 1 diabetes after transplantation into the kidney capsule of hiPSC-derived human islet-like organoids (n=1000), (dotted line with squares); human islets (n=1000), (dashed line); or mock treatment (solid line with white circles).  FIG. 4I  shows a bar graph illustrating that human islet-like mini organs (“human islets organoids”) generated by the methods described herein are able to secrete insulin postprandially. The graph of  FIG. 4I  shows the serum levels of human c-peptide (pmol/L) in NODSCID mice 4 weeks after the transplantations described in  FIG. 4H  under random fed (left bar), 8 hour fasted (middle bar), and refed (right bar) conditions. Human c-peptide levels provide a measure of insulin secretion from the transplants that is distinct from endogenous murine insulin. 
         FIGS. 5A-5E  are diagrams and plots showing the generation of functional islet-like organoids in 3D gellan gum cultures.  FIG. 5A  is a schematic describing the generation of hiPSCs stably incorporating dual reporters for insulin expression (GFP) and insulin secretion (luciferase).  FIG. 5B  is a bar graph showing the increased expression of human insulin during the differentiation of hiPSCs incorporating the dual reporters.  FIG. 5C  is a bar graph comparing the glucose stimulated insulin secretion (GSIS) from human islet-like organoids generated using the methods described herein with human islets. GSIS, as measured using secreted luciferase, in single islet-like organoids or human islets in response to 3 mM and 20 mM glucose. Arrows indicate functional organoids capable of increasing insulin secretion in response to a glucose challenge.  FIG. 5D  is a bar graph comparing the insulin secretion of independent batches of islet-like organoids, prepared as described herein, to mouse islets as a negative control. Insulin secretion was measured in response to 3 mM glucose (G3 mM), 20 mM glucose (G20 mM), 20 mM glucose (G20 mM) and Exendin-4 (Ex4), or 20 mM potassium chloride (KC120 mM) after 133 days in culture. Response was measured as secreted luciferase activity from pooled organoids (100 organoids/sample).  FIG. 5E  shows intracellular luciferase activity as a measure of intracellular insulin (100 organoids/sample). 
         FIGS. 6A-6H  are schematics and images showing generation of functional human mini-organs in a dish.  FIG. 6A  is a schematic showing generation of functional human mini organs including human islets, pancreas, liver, heart, and intestine.  FIGS. 6B-6C  show the generation of a human heart organoid.  FIG. 6B  (top) provides a schematic illustrating the protocol for differentiation of a human pluripotent stem cell (hPSC) into a cardiomyocyte (when cultured in 2D) or into a mini heart (when co-cultured with hADSCs and HUVECs in 3D).  FIG. 6B  (bottom left) provides a plot showing relative expression of cardiomyocyte-specific genes human MLC2a (hMLC2a), human Nkx2-5 (hNkx2-5), alpha myosin heavy chain (alphaMHC), and KCNQ1 before (day 0) and at day 18 of the differentiation protocol, with and without the PPARδ agonist GW501516. At the bottom right of  FIG. 6B  is a videomicrograph showing beating of the hiPSC-derived cardiomyocytes.  FIG. 6C  shows an image of a human mini heart-like organoid generated by culturing the hiPSC-derived cardiomyocytes with hADSC and HUVEC, as described in the schematic above.  FIGS. 6D-6F  show generation of a human liver organoid.  FIG. 6D  (top) provides a schematic illustrating the protocol for differentiation of a human pluripotent stem cell (hPSC) into hepatocytes.  FIG. 6D  (bottom left) provides a plot showing relative expression of hepatocyte-specific genes AFP, ALB, and Cyp3a7, during the differentiation of 6 independent preparations. At the bottom right of  FIG. 6D  is a micrograph showing hiPSC-derived hepatocytes.  FIG. 6E  is a set of microcrographs showing hiPSC-derived hepatocytes. Top panels show expression of Cyp7a1 (Cyp7a1-GFP reporter, left) and SREBP1c (SREBP1c-GFP reporter, right) in hiPSC-derived hepatocytes indicating functional maturation. The bottom panels show hiPSC-derived hepatocytes cultured with (right) or without (left) phosphatidic acid (PA) overnight. Higher magnification images shown in the bottom left reveal the accumulation of lipid droplets in the hepatocytes treated with phosphatidic acid.  FIG. 6F  shows a human mini liver-like organoid generated by culturing the hiPSC-derived hepatocytes with hADSCs and HUVECs in the gellan gum 3D culture system.  FIG. 6G  (top) provides a schematic illustrating the protocol for differentiation of human pluripotent stem cells (hPSCs) into intestinal organoids when co-cultured with hADSCs and HUVECs in the gellan gum 3D culture system. The lower images shows budding of the human intestinal organoid cultures, consistent with crypt-like structures and indicating functional organoids.  FIG. 6H  (top) provides a schematic illustrating the protocol for differentiation of human pluripotent stem cells (hPSCs) into a mini pancreas when co-cultured with hADSCs and HUVECs in the gellan gum 3D culture system. The middle panels show images of insulin positive β cells, marked by the expression of green fluorescent protein driven by the insulin promoter, and the equivalent light microscopy image of pancreatic organoids. Exocrine cells are the remaining unlabeled cells. The bottom panels show an image of a single pancreatic organoid in a 96 well plate. 
         FIG. 7  is a schematic showing modeling of human Type 2 diabetes in a dish. Generation of functional human organs such as a liver, pancreas, and islets provides new therapeutic strategies in drug-screening and modeling of human type 2 diabetes. For example, hepatic organoids with accumulated lipids can be used to study early insulin resistance phenotypes, while human islet-like organoids can be used to study β cell death seen in late stage type 2 diabetes. Inserted graph shows the response of islet-like organoids to increasing concentrations of human amyloid polypeptide (hIAPP). The increase in propidium iodide (PI) staining in G0/G1 stage cells indicates that hIAPP induces apoptosis in islet-like organoids. 
         FIGS. 8A and 8B  are schematics showing platforms for drug screening and the subsequent evaluation of potential candidates for human type 2 diabetes and pancreatic cancer.  FIG. 8A  shows a scheme for screening potential drugs for use in human type 2 diabetes or human pancreatic cancer tumorigenesis in a dish. Organoid cultures are exposed to appropriate stress (e.g. high levels of free fatty acids (FFAs), high glucose levels, or relevant cytokines) to induce disease-like phenotypes prior to screening drug libraries for compounds that reverse or diminish disease indications.  FIG. 8B  shows approaches to evaluate potential drug candidates for type 2 diabetes and human pancreatic cancer tumorigenesis/metastasis in mice. Mice transplanted with individual (e.g. pancreas) or combinations of human organoids (e.g., pancreas and liver) are exposed to appropriate disease-inducing stressors (e.g. high fat/high cholesterol (HF/HC) diet) prior to treatment with potential disease altering drug candidates. 
         FIG. 9  is a set of schematics and images showing a structure of a pancreas and pancreatic tissue and images of a pancreatic islet-like organoid and pancreatic organoid generated herein. The schematics on the left of  FIG. 9  depict the anatomy and structure of a pancreas (top) and pancreatic islets (bottom). The image on the top-right corner of  FIG. 9  shows an iPSC-derived pancreatic organoid with pancreatic islets (as marked by green fluorescent protein expression driven by the insulin promoter) and an exocrine/duct component as indicated. The image in the middle-right of  FIG. 9  shows the corresponding light microscopy image of the fluorescent image above. The image on the bottom-right corner of  FIG. 9  shows a single pancreatic islet organoid. 
         FIGS. 10A-10D  is a set of schematics, images, heatmaps, and bar graphs summarizing the generation of islet-like organoids from PSCs.  FIG. 10A  is a schematic describing the protocol to generate islet-like organoids by culturing in 3D in gellan gum.  FIG. 10B  is a series of images recording the growth and differentiation of hPSCs into pancreatic lineages in 3D Gellan gum cultures, as described herein. Insulin expression is indicated by the green fluorescence seen at day 21.  FIG. 10C  is a heatmap representation of changes in gene expression during the differentiation of hiPSCs into islet-like organoids.  FIG. 10D  is a set of bar graphs reporting the changes in relative expression of the pluripotency marker Nanog, the endocrine hormones insulin, somatostatin, and glucagon, and the β cell lineage marker Nkx6-1 in hiPSCs and two stem cell lines (HuES8 and H1ES) during differentiation as described in the methods herein. Gene expression was measured by qPCR. 
         FIGS. 11A and 11B  show a heatmap and graph illustrating the role of WNT proteins in the hADSCs in 3D culture.  FIG. 11A  is a heatmap depiction of gene expression changes in hADSCs during the spontaneous self-organization that occurs in 3D culture. Genes that are induced in the WNT5a pathway are listed.  FIG. 11B  is a graph showing the relative levels of several individual WNT proteins in hADSC 3D culture over time, identifying the WNT5a protein as the predominant protein expressed. 
         FIGS. 12A-12H  show a set of bar graphs and images illustrating the role of WNT proteins in the metabolic maturation of iPSC-derived islet organoids.  FIG. 12A  shows bar graphs comparing the expression of Fltp and Esrrg genes in iPSC-derived islet organoids (day 21, generated without co-culture with hADSCs or HUVECs) after treatment with PBS, WNT3a (500 ng/ml), WNT4 (100 ng/ml), or WNT5a (400 ng/ml) for 5 days.  FIG. 12B  is a bar graph showing the induction of Esrrg gene expression in hiPSC-derived islet organoids, generated in the absence of supporting hADSC or HUVECs, in response to increasing doses of WNT4 (0, 10, 25, 50, 100, 200 ng/ml) and WNT5a (0, 25, 50, 100, 200, 400 ng/ml).  FIG. 12C  is a bar graph showing the induction of mitochondrial genes involved in oxidative phosphorylation (Cox7a2, Ndufa1, Ndufa7), lactate dehydrogenase (Ldha) and Fltp (a Wnt/planar cell polarity (PCP) effector and reporter gene) in hiPSC-derived islet organoids, generated in the absence of supporting hADSC or HUVECs, in response to increasing doses of WNT4 (0, 10, 25, 50, 100, 200 ng/ml) and WNT5a (0, 25, 50, 100, 200, 400 ng/ml).  FIG. 12D  shows fluorescent images showing mitochondrial (Mitotracker; Mito-Red) and insulin (Insulin-GFP) levels in hiPSC-derived islet organoids (day 27) after 8 days treatment with PBS or WNT4 (100 ng/ml).  FIG. 12E  shows fluorescent images of FACS analysis of hiPSC-derived islet organoids (day 27) after 8 days treatment with PBS or WNT4 (100 ng/ml).  FIGS. 12F, 12G, and 12H  show a set of bar graphs illustrating the results of FACS analyses of hiPSC-derived islet organoids (day 27) after 8 days treatment with PBS, WNT4 (100 ng/ml), WNT5a (400 ng/ml), control fibroblast conditioned media (50%), or WNT5a secreting fibroblast conditioned media (50%). The WNT proteins used were recombinant human (rh) proteins. 
         FIG. 13  shows a bar graph demonstrating a role for WNT4 in the functional maturation of hiPSC-derived islet organoids. Human iPSC (hiPSC)-derived islet organoids (day 22) were treated with PBS (Vehicle, “Veh”) or WNT4 (100 ng/ml) for 12 days, and the secretion of human c-peptide was measured in response to low glucose βmM, “G3 mM”), high glucose (20 mM, “G20 mM”), or high KCl levels (20 mM, “KCL20 mM”). 
     
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     The invention features compositions and methods that are useful for generating scalable, functional, vascularized organoids in vitro, particularly human pancreatic or pancreatic islet organoids. The invention is based, at least in part, on the discovery that culturing iPSC-derived beta-like cells with human adipose-derived stem cells (hADSC) and human umbilical vein endothelial cells (Huvec) in a three-dimensional matrix containing gellan gum generated functional pancreatic and pancreatic islet organoids. 
     The organoids generated were vascularized and exhibited functional properties, such as glucose-stimulated insulin secretion (GSIS). Islet transplantation is known as the best therapy for curing insulin deficient diabetes such as type 1 and late stage of type 2 diabetes. Recent studies have shown the possibility of generating glucose responsive insulin producing beta-like cells from human Pluripotent Stem Cells (PSCs), however the generation of functional, vascularized pancreatic islets from PSCs capable of secreting insulin, glucagon and somatostatin in response to nutrients has not been previously achieved. 
     Studies described herein demonstrate that using the self-organizing function of human adipose-derived stem cells (hADSC), HUVEC, and human iPSC-derived beta-like cells allows for the in vitro generation of glucose-responsive insulin secreting islet-like organoids with the ability to form functional vasculature. Studies herein further demonstrate the successful scaling of islet-like organoids production through the use of Gellan gum based 3D culture systems. Using a Gaussia luciferase reporter to measure insulin secretion, the functional heterogeneity in hiPSC-derived islet-like organoids was characterized. Without intending to be bound by theory, results herein suggest that the novel human islet-like organoids may offer a therapeutic treatment for diabetes, as well as offer a platform for drug screening, genome editing, and the modeling of organogenesis and pathogenesis of diabetes. 
     Pancreas 
     In some aspects, the invention provides a pancreatic organoid or a pancreatic islet organoid. The pancreas is an organ that lies in the abdomen and has endocrine and exocrine functions.  FIG. 9  provides schematics showing the structure of the pancreas. The portion of the pancreas having an endocrine role are cell clusters called “pancreatic islets” (also known as islets of Langerhans). Pancreatic endocrine secretions include hormones that regulate glucose metabolism and blood glucose concentration. Four main cell types are present in the islets: alpha cells which secrete glucagon (a hormone that increases blood glucose concentration); beta cells which secrete insulin (a hormone that decreases blood glucose concentration); delta cells, which secrete somatostatin (a hormone that regulates alpha and beta cells), and gamma cells which secrete pancreatic polypeptide. 
     The portion of the pancreas that has an exocrine role is referred to as the exocrine component. The exocrine pancreatic secretions contain digestive enzymes that pass into the small intestine and help break down carbohydrates, proteins, and lipids. The exocrine component has ducts arranged in clusters called pancreatic acini. Pancreatic exocrine secretions are secreted into the lumen of the acinus, which accumulate and drain into the pancreatic duct and duodenum. 
     Pancreatic islet organoids and pancreatic organoids of the invention mimic the structure of a pancreatic islet and a pancreas, respectively. In some embodiments, the pancreatic islet organoid or pancreatic organoid of the invention contains any one or more of the following cells: an iPSC-derived beta-like cell, an iPSC-derived alpha-like cell, an iPSC derived delta-like cell, and an iPSC-derived duct-like cell. In some embodiments, the pancreatic organoid of the invention contains an iPSC-derived exocrine component. In some embodiments, the iPSC is a human iPSC (hiPSC). Human embryonic stem cells and human induced pluripotent stem cells are commercially available (e.g., from WiCell, which provides iPS(IMR-90)-1, iPS(IMR-90)-4 and iPS(Foreskin)-1). Human induced pluripotent stem cells can also be generated using methods known in the art from a variety of somatic cell types (Yu, J., K. Hu, et al. (2009). “Human induced pluripotent stem cells free of vector and transgene sequences.” Science 324(5928): 797-801). 
     Pancreatic islet organoids and pancreatic organoids of the invention also exhibit function(s) of a pancreatic islet and a pancreas. In certain embodiments, the pancreatic islet organoid or pancreatic organoid exhibits any one or more of the following functions: glucose-stimulated insulin secretion (GSIS), KCl-stimulated insulin secretion, GLP-1 stimulated insulin secretion, somatostatin secretion, and glucagon secretion. In some embodiments, the pancreatic islet or pancreatic organoid expresses any one or more of the transcription factors Pdx1, MafA, Pax4, Pax6, NeuroD1, Nkx6-1, Gata6, and Foxa2. 
     Generation of Pancreatic and Pancreatic Islet Organoids 
     In some other aspects, the invention features methods of generating a pancreatic or pancreatic islet organoid. Recent studies have shown that while it was possible to generate glucose responsive insulin producing beta-like cells, efforts to generate pancreatic islets which are capable of secreting insulin, glucagon and somatostatin in response to nutrients, as well as efforts to obtain vascularization from stem cells, have not succeeded. Described herein are results demonstrating that using the self-organizing function of human Adipose-derived stem cells (hADSC), human umbilical vein endothelial cells (HUVEC), and human iPSC-derived beta-like cells, glucose responsive insulin secreting islet-like organoids capable of functional vascularization are successfully generated in vitro. Further, islet-like organoid generation methods were successfully scaled up using gellan gum based 3D culture systems. The functional heterogeneity in hiPSC-derived human islet-like organoids was also investigated using a Gaussia luciferase reporter to measure insulin secretion. 
     Generation of functional human organs provides new therapeutic strategies in drug-screening, disease modeling and inhibiting or preventing end point organ failure. Efficient stepwise differentiation methods from human embryonic stem cells (hESC) and human induced pluripotent stem cells (hiPSC) to insulin producing β-like cells were demonstrated previously. D&#39;Amour et al and Kroon E et al reported the efficient differentiation of hESCs into insulin producing cells which, after 4 to 5 months in vivo maturation, are able to secrete insulin in response to glucose (D&#39;Amour et al., 2006, Nature biotechnology 24, 1392-1401; Kroon et al., 2008, Nature biotechnology 26, 443-452). Recently, Rezania et al. and Pagliuca et al. reported differentiation methods that induced formation of mature human beta-like cells in vitro in that they expressed terminal β-cells marker, MAFA and Nkx6-1 and exhibited partial functionality (e.g., insulin secretion) (Rezania et al., 2014, Nature Biotechnology Nov; 32(11):1121-33; Pagliuca et al., 2014, Cell 159, 428-439). However, in contrast to cadaveric human islets, those beta-like cells required in vivo functional maturation for a few months, and lacked the functionality provided by the other pancreatic islet cell types, such as glycemic control by α-cells (glucagon secrete) and δ-cells (somatostatin secretion). Further, the beta-like cells lacked both a mesenchyme and vascularized endothelial cells, which human islets naturally have. These crucial differences between hPSCs derived beta-like cells and human islets may compromise the ability of hPSCs based therapies to treat insulin dependent diabetes (such as type 1 or late stage type 2 diabetes). 
     Previously, it was identified that a metabolic transition occurs during the neonatal to adult maturation of β-cells in which the orphan nuclear receptor Estrogen-related receptor γ (ERRγ) regulates an increase in oxidative metabolism required for fully functional β cells. Consistent with this result, human iPSC-derived β like cells expressing insulin, MAFA, and Nkx6-1 can be metabolically matured through the overexpression of ERRγ to increase their oxidative metabolism and thereby enhance their glucose stimulated insulin secretion (GSIS) functionality. These results indicated that in addition to the expression of lineage determination factors such as PDX1, MAFA, Nkx6-1, and insulin, further cellular signaling which mature the β-cells&#39; metabolism is required to generate fully functional β-cells. 
     During early pancreas organogenesis, newly specified pancreatic cells originate from the foregut endodermal sheet and form a pancreatic bud, a condensed tissue mass that is soon vascularized. A similar progression has been observed in liver organogenesis as well. Such large-scale morphogenetic changes depend on the exquisite orchestration of signals between endodermal epithelial, mesenchymal, and endothelial progenitors before blood perfusion. Takebe et al, successfully generated hepatic organ buds by culturing hepatic endoderm cells with endothelial and mesenchymal linages which rapidly vascularized and functional matured in vivo (Takebe et al., 2013, Nature 499, 481-484). 
     Previous work did not reveal the possibility of generating in vitro other organoid tissue types, such as pancreas organoids, which were mature, functional, and vascularized. Further, previous work showed a lack of scalability because the organoids were generated using MATRIGEL® matrix, which is not efficient to use for scaled-up production. 
     Described herein are studies demonstrating successful large-scale generation of human islet-like organoids which are capable of secreting insulin and which are vascularized, as seen in human islets. It is demonstrated herein that (1) human adipose derived stem cells (hADSCs) have a self-organizing capacity; (2) late stage pancreatic progenitors are capable of forming an islet-like cluster when co-cultured with HUVECs and hADSCs with comparable efficiency to beta-like cells; (3) human islet-like organoids had improved expression of lineage determination factors as well as metabolic regulatory genes including ERRγ; (4) islet insulin secretion assays, measured using a Gaussia Luciferase proinsulin system, revealed that human islet-like organoids contain functional cells capable of secreting insulin in response to glucose; (5) human islet-like organoids exhibited vascularization; (6) human islet-like organoids derived from hiPSC by the method described herein recaptured human islet organogenesis and pathogenesis of type 1 and type 2 diabetes in a dish; (7) human islet-like organoids derived from hiPSC by the method described herein offered a new replaceable resource for human islet transplantation to treat type 1 and type 2 diabetes; and (8) human islet-like organoids transplanted into an STZ-induced NODSCID mouse model of type 1 diabetes ameliorated type 1 diabetes in the recipient animals. ( FIGS. 4H and 4I ). 
     Also described herein are studies in which the role of certain Wnt (also “WNT” herein) proteins was assessed in developing human islet-like organoids which are capable of secreting insulin and which are vascularized, as seen in human islets. The WNT gene family consists of structurally related genes that encode secreted signaling proteins, which have been implicated in oncogenesis and in several developmental processes, including regulation of cell fate and patterning during embryogenesis. Wnt proteins comprise a major family of signaling molecules that orchestrate and influence a variety of cell biological and developmental processes. Wnt proteins undergo a complex set of posttranslational modifications involving several highly specialized processing enzymes. Upon release from the cell, the Wnt proteins interact with a number of molecules in the extracellular environment, such as glycans, protein-binding partners (e.g., WIF, Sfrp) and cell surface receptors. (Willert, K. et al., 2012, Cold Spring Harbor,  Perspectives in Biology,  2012). It is demonstrated herein that (1) Wnt5a is the predominant Wnt protein that induces the self-organization of hADSCs ( FIGS. 11A and 11B ); (2) Wnt5a, as well as Wnt4, activate the ERRγ-mitochondrial metabolic pathway ( FIGS. 12A-12H ); (3) Wnt4 is sufficient to induce in vitro functional maturation of hiPSC-derived islet-like organoids in the absence of additional cell types such as hADSC and HUVECs ( FIG. 13 ). 
     Methods of Treatment 
     Islet transplantation is a therapy for treating insulin deficient diabetes such as type 1 and late stage type 2 diabetes. Thus, in another aspect, the present invention provides methods of treating a pancreatic disease such as type 1 or type 2 diabetes comprising administering a pancreatic or pancreatic islet organoid of the invention to a subject (e.g., a mammal such as a human) by transplantation. One embodiment is a method of treating a subject suffering from or susceptible to a pancreatic disease (e.g., type 1 diabetes) or disorder or symptom thereof. The method includes the step of transplanting a pancreatic or pancreatic islet organoid of the invention to the mammal sufficient to treat the disease or disorder or symptom thereof, under conditions such that the disease or disorder is treated. 
     As used herein, the terms “treat,” treating,” “treatment,” and the like refer to reducing or ameliorating a disorder and/or symptoms associated therewith. It will be appreciated that, although not precluded, treating a disorder or condition does not require that the disorder, condition or symptoms associated therewith be completely eliminated. 
     As used herein, the terms “prevent,” “preventing,” “prevention,” “prophylactic treatment” and the like refer to reducing the probability of developing a disorder or condition in a subject, who does not have, but is at risk of or susceptible to developing a disorder or condition. 
     The therapeutic methods of the invention (which include prophylactic treatment) in general comprise administration (in particular, transplantation) of an effective amount of a pancreatic or pancreatic islet organoid to a subject (e.g., animal, human) in need thereof, including a mammal, particularly a human. The administration of the pancreatic or pancreatic islet organoid may be by any suitable means that results in an amount of the organoid that, combined with other components, is effective in ameliorating, reducing, or stabilizing a pancreatic disease such as type 1 or type 2 diabetes. 
     In some aspects, the subject is further administered an immunosuppressant. The immunosuppressant can be administered to the subject before, during, or after the subject is administered (e.g., transplanted) with the organoid. The immunosuppressive agent can be an agent that inhibits or prevents rejection (e.g., acute rejection) of the transplanted organoid upon transplantation, or an agent that maintains immunosuppression after the transplantation. Immunosuppressants include, but are not limited to, basilizimab, antithymocyte globulin, alemtuzumab, prednisone, azathioprine, mycophenolate, cyclosporine, sirolimus, and tacrolimus. 
     In some embodiments, at least about 100,000, at least about 200,000, at least about 300,000, at least about 400,000, at least about 500,000, at least about 600,000, at least about 700,000, at least about 800,000, at least about 900,000 or at least about 1 million pancreatic islet organoids are transplanted into the subject. In some embodiments, islets of the subject are removed prior to transplanting the organoids of the invention. In some other embodiments, pancreatic islet organoids are transplanted into a subject by injection into the upper abdomen of the subjects. In some embodiments, the pancreatic islet organoids are injected into the liver. The pancreatic islet organoids can be injected into the subject using a catheter. In some other embodiments, the pancreatic organoid or pancreatic islet organoid is administered to the subject by surgery. In another embodiment, pancreatic islet organoids are transplanted onto the omentum. For omentum transplantation, a layering technique can be used in which the islet organoid (or cells thereof) are combined with autologous plasma and are laparoscopically layered onto the omentum. A solution (20 ml) containing recombinant thrombin (1000 U/ml) is next layered over the islet organoid, followed by another layer of autologous plasma to produce a biodegradable biologic scaffold that can survive and function in the patient for at least a year (See, e.g., Baidal, D. et al., 2017 , N. Engl. J. Med.,  376:19). In another embodiment, hydrogel biomaterials that mitigate an immune response by the recipient can be used for islet organoid transplantation. (See, e.g., Vegas, A. et al., 2016 , Nature Biotechnology,  34:345-352). 
     To further reduce an immune reaction to the transplanted organoid in the subject, the organoid can be encapsulated in a hydrogel and then transplanted in the subject. Such methods of transplantation are further described in Vegas et al., Nature Medicine 2016, doi:10.1038/nm.4030; Vegas et al., Nature Biotechnology 2016, doi:10.1038/nbt.3462. In some embodiments, the hydrogel contains an alginate or alginate derivative (e.g., triazole-thiomorpholine dioxide). Various modifications of alginate hydrogels that substantially reduce inflammatory or fibrotic effects of alginate hydrogels have also been identified (Vegas et al., Nature Biotechnology 2016, doi:10.1038/nbt.3462). Thus, in some other embodiments, the hydrogel contains a chemical modification that reduces an inflammatory effect of the transplanted organoid in the subject. 
     Screening Assays 
     Pancreatic islet organoids and pancreatic organoids of the invention can be useful for modeling diseases of the pancreas in vitro or in vivo. Such pancreas disease models can be used to identify drugs that are useful for treatment of a pancreatic disease. Thus, in some aspects, the invention provides methods for identifying modulators, i.e., candidate or test compounds or agents (e.g., proteins, peptides, peptidomimetics, peptoids, polynucleotides, small molecules or other drugs) that are useful for the treatment of a pancreatic disease, particularly type 2 diabetes and/or pancreatic cancer. In one embodiment, the agent modulates an activity of an organoid of the invention. 
     The test agents of the present invention can be obtained singly or using any of the numerous approaches in combinatorial library methods known in the art, including: biological libraries; peptoid libraries (libraries of molecules having the functionalities of peptides, but with a novel, non-peptide backbone which are resistant to enzymatic degradation but which nevertheless remain bioactive; see, e.g., Zuckermann, R. N. (1994) et al., J. Med. Chem. 37:2678-85); spatially addressable parallel solid phase or solution phase libraries; synthetic library methods requiring deconvolution; the ‘one-bead one-compound’ library method; and synthetic library methods using affinity chromatography selection. The biological library and peptoid library approaches are limited to peptide libraries, while the other four approaches are applicable to peptide, non-peptide oligomer or small molecule libraries of compounds (Lam (1997) Anticancer Drug Des. 12:145). 
     Examples of methods for the synthesis of molecular libraries can be found in the art, for example in: DeWitt et al. (1993) Proc. Natl. Acad. Sci. U.S.A. 90:6909; Erb et al. (1994) Proc. Natl. Acad. Sci. USA 91:11422; Zuckermann et al. (1994) J. Med. Chem. 37:2678; Cho et al. (1993) Science 261:1303; Carrell et al. (1994) Angew. Chem. Int. Ed. Engl. 33:2059; Carell et al (1994) Angew. Chem. Int. Ed. Engl. 33:2061; and Gallop et al. (1994) J. Med. Chem. 37:1233. 
     Libraries of compounds may be presented in solution (e.g., Houghten (1992), Biotechniques 13:412-421), or on beads (Lam (1991), Nature 354:82-84), chips (Fodor (1993) Nature 364:555-556), bacteria (Ladner, U.S. Pat. No. 5,223,409), spores (Ladner U.S. Pat. No. 5,223,409), plasmids (Cull et al. (1992) Proc Natl Acad Sci USA 89:1865-1869) or on phage (Scott and Smith (1990) Science 249:386-390; Devlin (1990) Science 249:404-406; Cwirla et al. (1990) Proc. Natl. Acad. Sci. 87:6378-6382; Felici (1991) J. Mol. Biol. 222:301-310; Ladner supra.). 
     Chemical compounds to be used as test agents (i.e., potential inhibitor, antagonist, agonist) can be obtained from commercial sources or can be synthesized from readily available starting materials using standard synthetic techniques and methodologies known to those of ordinary skill in the art. Synthetic chemistry transformations and protecting group methodologies (protection and deprotection) useful in synthesizing the compounds identified by the methods described herein are known in the art and include, for example, those such as described in R. Larock (1989) Comprehensive Organic Transformations, VCH Publishers; T. W. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 2nd ed., John Wiley and Sons (1991); L. Fieser and M. Fieser, Fieser and Fieser&#39;s Reagents for Organic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995), and subsequent editions thereof. 
     Combinations of substituents and variables in compounds envisioned by this invention are only those that result in the formation of stable compounds. The term “stable”, as used herein, refers to compounds which possess stability sufficient to allow manufacture and which maintains the integrity of the compound for a sufficient period of time to be useful for the purposes detailed herein (e.g., transport, storage, assaying, therapeutic administration to a subject). 
     The compounds described herein can contain one or more asymmetric centers and thus occur as racemates and racemic mixtures, single enantiomers, individual diastereomers and diastereomeric mixtures. All such isomeric forms of these compounds are expressly included in the present invention. The compounds described herein can also be represented in multiple tautomeric forms, all of which are included herein. The compounds can also occur in cis- or trans- or E- or Z-double bond isomeric forms. All such isomeric forms of such compounds are expressly included in the present invention. 
     Test agents of the invention can also be peptides (e.g., growth factors, cytokines, receptor ligands) or polynucleotides encoding such peptides. 
     Screening methods of the invention identify agents that increase or decrease a biological activity of pancreatic islet organoids and pancreatic organoids of the invention. In some embodiments, a pancreatic disease, such as type 2 diabetes or pancreatic cancer, is induced or mimicked in the pancreatic islet organoid or pancreatic organoid. Type 2 diabetes in the pancreatic islet or pancreatic organoid can be induced, for example, by contacting the organoid with free fatty acids (FFAs), glucose, and cytokines (in particular, high levels of glucose and/or high levels of FFAs). In one embodiment, a pancreatic organoid is co-cultured with pancreatic cancer cells, stellate cells and immune cells to create a human pancreatic cancer microenvironment in vitro. 
     In some embodiments, the organoid is contacted with a candidate agent, and an effect of the candidate agent on a biological activity, function, or event is assayed. In some embodiments, the candidate agent is a drug approved by the Food and Drug Administration (FDA). For example, biological activities of a pancreatic islet organoid or pancreatic organoid assayed in the screening methods of the invention include insulin secretion (e.g., glucose-stimulated insulin secretion (GSIS)), beta cell apoptosis, LDHA activity, K(ATP) channel activity, mitochondrial function, level or activity of NDUFA4, ESRRG, KCNK3, or MAFA polypeptide or polynucleotide, cell death, cell growth, and metastasis. In some embodiments, the agent increases GSIS. 
     In some other embodiments, an organoid of the invention (e.g., pancreatic islet organoid or pancreatic organoid) is transplanted into a host to model pancreatic disease, such as type 2 diabetes or pancreatic cancer, in vivo. Methods of transplanting an organ or organoid are known in the art. The host can be any non-human mammal, such as a rat or mouse. 
     To reduce an immune reaction to the transplanted organoid in the host after, the organoid can be encapsulated in a hydrogel and then transplanted in the host. Such methods of transplantation are further described in Vegas et al., Nature Medicine 2016, doi:10.1038/nm.4030; Vegas et al., Nature Biotechnology 2016, doi:10.1038/nbt.3462. In some embodiments, the hydrogel contains an alginate or alginate derivative (e.g., triazole-thiomorpholine dioxide). Various modifications of alginate hydrogels that substantially reduce inflammatory or fibrotic effects of alginate hydrogels have also been identified (Vegas et al., Nature Biotechnology 2016, doi:10.1038/nbt.3462). Thus, in some other embodiments, the hydrogel contains a chemical modification that reduces an inflammatory effect of the transplanted organoid in the host. 
     In some embodiments, a pancreatic organoid and liver organoid are co-transplanted in the host. The liver is a major target organ for metastasis of pancreatic cancer. In mice in vivo endothelial cells in the mini pancreas and in the mini liver are connected to each other and create a pancreas-liver vasculature network for pancreatic cancer metastasis. Therefore, a host co-transplanted with a pancreatic organoid and liver organoid can be useful for studies of human pancreatic cancer metastasis into human liver. 
     In some embodiments, the host transplanted with an organoid of the invention is administered an environmental stress (e.g., administered a high fat/high glucose diet or administered pancreatic cancer cells) to induce or mimic a pancreatic disease in the host. In some other embodiments, the host is transplanted with a pancreatic islet or pancreatic organoid and/or a liver organoid where a disease (e.g., type 2 diabetes or pancreatic cancer) has been induced. 
     In some embodiments, the host is administered with a candidate agent. In certain embodiments, the candidate agent is a drug approved by the Food and Drug Administration (FDA). In some embodiments, an effect of the candidate agent on a host phenotype (such as biological activity or function associated with the pancreas, or activities associated with a disease) is assayed. Exemplary biological activities include insulin secretion (e.g., glucose-stimulated insulin secretion (GSIS)), beta cell apoptosis, LDHA activity, K(ATP) channel activity, mitochondrial function, level or activity of NDUFA4, ESRRG, or MAFA polypeptide or polynucleotide, cell death, cell growth, and metastasis. In some embodiments, the agent increases GSIS. 
     In any one of the embodiments herein, the effect of the candidate agent (i.e., ability to modulate a pancreatic activity or function) is measured relative to a reference. The reference can be, for example, an untreated pancreatic islet organoid or pancreatic organoid. In some embodiments, the reference is a host transplanted with an organoid of the invention, where the host is not administered with a candidate agent. 
     Agents useful in the methods of the invention can also be detected by identifying an increase in expression of a desirable marker (e.g., MAFA as a beta cell fate marker). The level of expression can be measured in a number of ways, including, but not limited to: measuring the mRNA encoded by the genetic markers; measuring the amount of protein encoded by the genetic markers; or measuring the activity of the protein encoded by the genetic markers. 
     The level of mRNA corresponding to a marker can be determined both by in situ and by in vitro formats. The isolated mRNA can be used in hybridization or amplification assays that include, but are not limited to, Southern or Northern analyses, polymerase chain reaction analyses and probe arrays. In one format, mRNA (or cDNA) is immobilized on a surface and contacted with the probes, for example by running the isolated mRNA on an agarose gel and transferring the mRNA from the gel to a membrane, such as nitrocellulose. In an alternative format, the probes are immobilized on a surface and the mRNA (or cDNA) is contacted with the probes, for example, in a two-dimensional gene chip array described below. A skilled artisan can adapt known mRNA detection methods for use in detecting the level of mRNA encoded by the markers described herein. 
     The level of mRNA in a sample can be evaluated with nucleic acid amplification, e.g., by rtPCR (Mullis (1987) U.S. Pat. No. 4,683,202), ligase chain reaction (Barany (1991) Proc. Natl. Acad. Sci. USA 88:189-193), self-sustained sequence replication (Guatelli et al. (1990) Proc. Natl. Acad. Sci. USA 87:1874-1878), transcriptional amplification system (Kwoh et al. (1989) Proc. Natl. Acad. Sci. USA 86:1173-1177), Q-Beta Replicase (Lizardi et al. (1988) Bio/Technology 6:1197), rolling circle replication (Lizardi et al., U.S. Pat. No. 5,854,033) or any other nucleic acid amplification method, followed by the detection of the amplified molecules using techniques known in the art. As used herein, amplification primers are defined as being a pair of nucleic acid molecules that can anneal to 5′ or 3′ regions of a gene (plus and minus strands, respectively, or vice-versa) and contain a short region in between. In general, amplification primers are from about 10 to 30 nucleotides in length and flank a region from about 50 to 200 nucleotides in length. Under appropriate conditions and with appropriate reagents, such primers permit the amplification of a nucleic acid molecule comprising the nucleotide sequence flanked by the primers. 
     The practice of the present invention employs, unless otherwise indicated, conventional techniques of molecular biology (including recombinant techniques), microbiology, cell biology, biochemistry and immunology, which are well within the purview of the skilled artisan. Such techniques are explained fully in the literature, such as, “Molecular Cloning: A Laboratory Manual”, second edition (Sambrook, 1989); “Oligonucleotide Synthesis” (Gait, 1984); “Animal Cell Culture” (Freshney, 1987); “Methods in Enzymology” “Handbook of Experimental Immunology” (Weir, 1996); “Gene Transfer Vectors for Mammalian Cells” (Miller and Calos, 1987); “Current Protocols in Molecular Biology” (Ausubel, 1987); “PCR: The Polymerase Chain Reaction”, (Mullis, 1994); “Current Protocols in Immunology” (Coligan, 1991). These techniques are applicable to the production of the polynucleotides and polypeptides of the invention, and, as such, may be considered in making and practicing the invention. Particularly useful techniques for particular embodiments will be discussed in the sections that follow. 
     The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the assay, screening, and therapeutic methods of the invention, and are not intended to limit the scope of what the inventors regard as their invention. 
     EXAMPLES 
     Example 1: Generation and Characterization of Pancreatic and Pancreatic Islet Organoids 
     Although an animal disease model can yield insight into the pathogenesis of diseases, drugs identified from screens using animal models often fail to be adopted in human patients. Generation of functional human organoids provides a new therapeutic strategy in drug-screening and disease modeling ( FIG. 1 ). Described herein is a novel technique to generate 3D human “mini-organs” or organoids in a dish. Using this technique, diseases such as human type 2 diabetes can be modeled in a dish to find effective drugs in genetic, patient or environmental specific diseases such as human type 2 diabetes. 
     Developing Gellan Gum Based 3D Culture System for β-Like Cells Differentiation 
     It is known that 3 dimensional (3D) culture systems contribute to facilitating self-organization and integration of cells. Therefore, MATRIGEL® matrix containing extracellular matrix components such as collagen and fibronectin is often used as the basement of a 3D culture system. However, MATRIGEL® matrix-based 3D culture systems are not ideal for large-scale human organoid generation because of their cost and difficulties in scale up. Described herein are Gellan-gum based 3D culture systems and methods for β-like cell differentiation, which are cost effective and easily scalable. Using a fully chemically-defined stepwise differentiation protocol ( FIG. 10A ) human pluripotent cells (hPSCs) are differentiated into insulin producing islet-like spherical cell clusters with high efficiency and reproducibility in Gellan-gum based 3D culture systems ( FIG. 10B ). Single dissociated pluripotent stem cells (PSCs) successfully formed into spheres within 5 days in Gellan gum containing STEMCELL™ TeSR™ media. Fifteen (15) to 21 days after differentiation in Gellan gum-containing Custom TeSR™ with defined small molecule stimulation, insulin positive GFP clusters were observed ( FIG. 10B ). Global transcriptome analysis by RNA-seq revealed the stepwise differentiation of hiPSCs into insulin positive cells expressing β cell lineage specific marker genes including Pdx1, Nkx6-1, GATA6 and MAFB ( FIG. 10C ). The differentiation of hiPSCs, as well as the human ESC lines HuES8 and H1ES, into islet-like cell clusters was further confirmed by the progressive loss of the pluripotent marker Nanog, the induction of the β cell specific marker Nkx6-1, and the progressive induction of the endocrine hormones insulin, somatostatin and glucagon, as determined by qPCR ( FIG. 10D ). These results demonstrate that the Gellan-gum based 3D culture systems is suitable for the generation of large-scale islet-like organoids from hPSCs. 
     Generation of Scalable, Human Islet-Like Organoids In Vitro 
     β-like cells derived from human embryonic stem cells (hESC) or human induced pluripotent stem cells (hiPSC) have limited functionality and lack the morphological and functional feature of human islets. Previous studies revealed that co-culturing hiPSC derived hepatocyte with human umbilical vein endothelial cells (HUVECs) and human bone marrow-derived mesenchymal stem cells (hMSC) generates self-organized 3D liver-bud spheres in matrigel (Takebe et al., 2013, Nature 499, 481-484). This study found that the liver “organoids” had superior expression of lineage determinant factors compared to the differentiation of isolated hepatocytes and that these organoids rapidly vascularized and functionally matured in vivo. 
     Studies herein found that hiPSC-derived pancreatic progenitor cells (hiPSC-PP) generated using a 2D differentiation protocol (Yoshihara et al, Cell Metab. 23, 622-634) did not self-organize in 3D MATRIGEL® matrix ( FIG. 2A ). In contrast, HUVEC cells rapidly formed a vasculature-like structure while human adipocyte-derived stem cells (hADSCs) self-organized in 3D MATRIGEL® matrix ( FIG. 2A ). In MATRIGEL® matrix, dispersed hADSC cells projected processes within 4 hours, formed a cloth-like wrapper within 12 hours, and adopted a sphere-like formation within 24 to 48 hours ( FIG. 2B ). Furthermore, a minimum cell density for self-organization was identified (i.e., ˜10,000-20,000 cells in 300 μl of MATRIGEL® matrix in ˜2 cm 2  well ( FIG. 2C ). RNA-seq analysis identified dynamic transcriptional changes during hADSC 3D self-organization, suggesting that the ability to self-organize under 3D culture conditions is an inherent feature of naïve hADSCs ( FIG. 2D ). These results identify the mesenchymal hADSC as a resource for generating self-organizing organoids. 
     To explore pancreatic organogenesis, hiPSC-PP (1×10 6  cells) cells were co-cultured with HUVECs (7×10 5  cells) and hADSCs (1-2×10 5  cells) ( FIG. 2E ) in Matrigel matrix. This co-culture yielded macroscopically visible 3D cell clusters 48 hours after seeding ( FIG. 2F ). Furthermore, insulin expression, based on the expression of a GFP reporter, was detected 5 days after seeding and increased with time in culture in the human islet-like organoids. In addition, HUVECs-based endothelial cells are integrated inside the organoids as shown by fluorescence-labeled (mCherry) HUVECs ( FIG. 2G ). 
     The limitations of MATRIGEL® matrix for organoid production include high cost, difficult organoid recovery, scaling restrictions, and batch to batch variabilities. 
     Described herein are methods to generate morphologically identical human islet-like organoids using gellan gum based 3D cultures ( FIGS. 4A-4G ).  FIG. 4A  shows a scheme for generation of functional, vascularized human pancreatic islets in as dish. Human induced pluripotent stem cells derived—pancreatic progenitors (hiPSC-PPs) (1×10 8  cells) were cultivated with a stromal cell population such as human umbilical vein endothelial cells (HUVECs) (2-7×10 6  cells) and human adipose-derived stem cells (hADSCs) (2-7×10 6 ) in 50 ml of gellan gum based 3D culture media ( FIG. 4B ).  FIG. 4B  shows that hiPSC-PP rapidly formed isle-like sphere formation with HUVECs and hADSCs within 5 days after seeding into the gellan gum based 3D culture media. Human islets like mini-organs expressed human insulin GFP reporter in 5 days after seeding with gradually enhancing GFP intensity. Co-culturing hiPSC-PP, hADSCs, and HUVECs according to this method, generated human islet-like organoids with high reproducibility that were morphologically similar to human islets ( FIG. 4C ). In addition, the generated human islet-like organoids contained insulin granules in β-like cells ( FIG. 4B ). Genes expression analyses revealed increased expression of f3 cell fate determinant genes (Insulin, Nkx6-1, PCSK1 and UCN3) and mitochondrial related metabolic genes (Esrrg, Ndufa1, Ndufa 12, Cox7a2. Atp5b) in the insulin expressing cell population (GFP enriched (GFP+)) in islet-like organoids compared to those prepared without hADSC and HUVEC co-culture ( FIG. 4D &amp; 4E ). Glucose-stimulated human c-peptide secretion assay revealed that islet-like organoids generated by this method are able to secrete human c-peptide in response to high (20 mM) glucose ( FIG. 4F ). 
     An in vitro functional vascularization test was then performed.  FIG. 4G  shows in vitro functional vascularization tests performed. Islet-like mini organs generated in gellan gum were transferred to MATRIGEL® matrix and cultured in endothelial growth media (EGM). Green fluorescence indicates expression of insulin genes. Within 24 hours to 48 hours after stimulation by EGM, the outgrowth of HUVEC cells was observed, indicating that human islet-like organoids generated by the method described herein possessed the ability to form vascular structures. 
     Establishment of Single Islet Insulin Secretion Assay Using Proinsulin-NanoLuc Gaussia Luciferase Assay System 
     It was previously published that a reporter construct, in which the Gaussia luciferase is placed within the c-peptide portion of proinsulin accurately measures insulin secretion without affecting β-cell function (Burns et al., 2015, Cell metabolism 21, 126-137). Using a lentiviral system, INS-1 cells stably expressing this Gaussia luciferase were generated ( FIGS. 3A-3F ). Luciferase secretion from INS-1 cells stably expressing Proinsulin-NanoLuc increased with high-glucose (20 mM), high glucose with Exendin-4 (G20 mM+Ex4), and the depolarizing agent, potassium chloride ( FIGS. 3A-3B ), confirming the utility of this reporter system. Next, the usefulness of this reporter to measure insulin secretion in mouse or human islets transiently infected with the Proinsulin-NanoLuc reporter was evaluated. Luciferase secretion in response to 20 mM high glucose was detected in both transiently infected mouse and human islets were detected ( FIGS. 3C-3F ). Importantly, the assay sensitivity was sufficient that insulin secretion could be qualified at the level of single islets. These results indicate that the Proinsulin-NanoLuc luciferase reporter based insulin secretion assay is applicable to not only the rat beta cell line INS-1 cells, but also to primary mouse and human primary β cells. 
     Establishment of hiPSC and hESC Cells Incorporating Dual Lineage and Functional Reporters 
     Human iPSCs and hESCs stably expressing reporters for βcell lineage (human insulin reporter) and β cell function (proinsulin-NanoLuc reporter) were generated, hiPSC hINS-GFP/Sec-Luc  and hESC hINS-GFP/Sec-Luc , respectively ( FIG. 5A ). First, a neomycin resistant construct of human insulin GFP reporter was generated by inserting human insulin promoter sequence of pGreenZeo lenti-reporter (SR10028PA-1, System Bioscience) into pGreenFire Lenti-Reporter plasmid (TR019PA-1, System Bioscience) (named as hINS-GFP-EF1a-Neo). hINS-GFP-EF1a-Neo lenti virus was infected into hiPSC and hESC by spin fection (800 g, 1 hour, 37 degree) followed by a media changed to fresh STEMCELL™ TeSR™ media. Three (3) days after the first infection, the cells were treated with 100 μg/ml G418 in STEMCELL™ TeSR™ media for 7 days. Selected hiPSC and hESC cells stably expressing hINS-GFP-EF1a-Neo were subsequently infected with the Proinsulin-NanoLuc (Addgene, Plasmid #62057)  lenti -virus by spin fection (800 g, 1 hour, 37 degree) followed by a media change to fresh STEMCELL™ TeSR™ media. Three (3) days after the second infection, the cells were treated with 5 μg/ml blasticysin and 100 m/ml G418 in STEMCELL™ TeSR™ media for 7 days. Subsequently, cells were maintained in STEMCELL™ TeSR™ media ( FIG. 5A ). The generated stable cell lines incorporating the dual reporters maintained self-renewal and pluripotency capabilities, as well as the capacity to differentiate into insulin producing β like cells ( FIG. 5B ). 
     Pooled Human Islet-Like Organoid Cultures Display Consistent Insulin Secretion Despite Variable Functionality Seen in Individual Organoids. 
     Recent studies have reported the generation of insulin producing β-like cells from hESC and hiPSC capable of secreting insulin in response to glucose (Pagliuca et al. 2014, Cell 159, 428-439; Rezania et al., 2014, Nature Biotechnology Nov; 32(11):1121-33; Russ et al., 2015, EMBO Journal 34, 1759-1772). However, fully functional human islet-like clusters able to appropriately secrete insulin in response to nutritional signals including glucose, amino acids, fatty acids and incretins such as GLP-1 have yet to be demonstrated. To date efforts have focused on the independent generation of insulin producing β-like cells, glucagon producing α-like cells, and somatostatin producing δ-like cells from hPSC. However, these approaches lack the supporting cells important for regulation, such as mesenchymal cells, adipose cells, and vasculature cells. Since the 3D structure of islets naturally enhances their function, these missing cellular components may compromise the functionality of islet-like cells clusters. In addition, organogenesis of pancreatic islets involves clonal expansion of β-cells, suggesting that these cells may have multiple functions in islet-like organoids. To test this idea, single organoid proinsulin secretion assays were performed. Human islet-like organoids generated by methods described herein are morphologically identical with human islet ( FIG. 4C ). However, significant variability was seen in the glucose-stimulated insulin secretion (GSIS) capabilities of individual human islet-like organoids compared to human islets, as measured by proinsulin luciferase secretion assay ( FIG. 5C ) Consistent GSIS functionality was demonstrated in pooled organoids (10 to 100 organoids for assay) ( FIG. 5D ). Furthermore, pooled human islet like organoids demonstrate enhanced GSIS when co-stimulation with GLP-1, as well as robust KCl-stimulated insulin secretion ( FIG. 5D ). 
     In vitro cultured iPSC-derived human pancreatic islet-like organoids generated herein retained their ability to respond to glucose, GLP1 and KCl after extended time (133 days) in culture ( FIG. 5D ). 
     Example 2: Generation and Characterization of Human Organoids 
     Functional human mini organs, including human islets, pancreas, liver, heart, and intestine, can be generated using the methods described herein ( FIG. 6A ).  FIG. 9  shows a structure of a human pancreas and human pancreatic islets within the pancreas. Using the methods herein, human pancreatic islets mini organs or organoids were generated in about 30 days. The pancreatic islets generated contained human induced pluripotent stem cell (hiPSC)-derived beta cells, alpha cells, delta cells, duct cells, as well as endothelial cells and hADSCs. The pancreatic islet organoids generated express key beta cells transcription factors such as Insulin, Nkx6-1, PCSK1, and UCN3, as well as key mitochondrial metabolic genes including Esrrg, Ndufa 1, Ndufa 12, Cox7a2 and Atp5b ( FIGS. 4D-4E ). The pancreatic islet organoids exhibited at least partial GSIS, KCl-stimulated insulin secretion, GLP-1 stimulated insulin secretion, vascularization, somatostatin secretion, and glucagon secretion ( FIGS. 4G, 5D ). 
     A human mini pancreas or human pancreatic organoid was generated in about 30 days ( FIG. 6H ). The human pancreatic organoid contained hiPSC-derived islets clustered within the interior of the organoid and a hiPSC-derived exocrine component surrounding the islets ( FIG. 6H ). The pancreatic organoid also contained endothelial cells and hADSC. Function of the human pancreatic organoid was demonstrated using an amylase secretion test, among other tests. 
     A human mini liver was generated in about 15 days ( FIGS. 6D-6F ). The human mini liver contained hiPSC derived hepatocytes, endothelial cells, and hADSCs. Analysis of functional characteristics of the human mini liver revealed that the mini liver expressed AFP, ALB, and Cyp3a7 ( FIG. 6D ), as well as the mature hepatocyte marker Cyp7a1 and lipogenesis marker SREBP1c ( FIG. 6E ). The human mini liver also exhibited insulin signaling, insulin resistance by palmitic acids, and lipid accumulation. The human mini liver is further tested for gluconeogenesis and metabolic function for drug metabolism. 
     A human mini heart was generated in about 15 days ( FIGS. 6B-6C ). The human mini heart contained hiPSC derived cardiomyocytes, endothelial cells, and hADSCs. The human mini heart expressed key cardiomyocyte genes such as hMlc2a, hNkx2-5, alpha MHC and KCNQ1 ( FIG. 6B ), and was seen to beat in cultures. 
     A human mini intestine was generated in about 30 days ( FIG. 6G ). The human mini intestine contained hiPSC derived intestinal cells, endothelial cells, and hADSCs. The human mini intestine expressed small intestine markers CDX2, Muc2, and Lgr5, and exhibited budding of intestinal organoids in response to R-Spondin. 
     Example 3: Transplantation of Functional Pancreatic Islet Organoids Rescued Type 1 Diabetic Mice 
     Expression of specific functional islets marker such as MAFA, UCN3 and mitochondrial oxidative genes such as ERRγ (Esrrg), Ndufa 1, Ndufa 12, Cox7a2 and Atp5b in hiPSC-derived human islet-like organoids was observed. Notably, these islet-like organoids recapture both human islets development as well as the pathogenesis of diabetes in a dish. Transplantation of these functional islet-like organoids rescue type 1 diabetic mice with long survival, rapid vascularization, and reduced immune rejection. 
     Example 4: Drug Screening and Disease Modeling in Human Islet-Like Organoids 
     Generation of functional human organs according to methods described herein provides new strategies for drug-screening and disease modeling. Specifically, functional organoids can be used as models of type 2 diabetes for drug screening ( FIG. 7 ). Human islet-like organoids responded to amyloid polypeptide (hIAPP) toxicity, an inducer of β cell loss in type 2 diabetic patients and islet dysfunction after transplantation in hyperglycemic patients, hIAPP dose-dependently induced G0/G1 arrest in 24 hours in human islet-like organoids ( FIG. 7 ). 
       FIG. 8A  provides a schematic showing experimental approaches to model type 2 diabetes and potentially screen for drugs using pancreatic islet and/or pancreatic organoids generated by the methods herein. In an exemplary assay, 3D mini organs are exposed to stressors that induce type 2 diabetes, such as high levels of free fatty acids (FFAs) and/or, glucose and selected cytokines. The stressed 3D mini organs are then treated with various drugs. In some embodiments, the drug is approved by the Food and Drug Administration (FDA). 
     As output, the following are assayed in human pancreatic islet organoids: insulin secretion, beta cell apoptosis (PI stain), lactate dehydrogenase A (LDHA) expression via a luciferase reporter, and changes in expression of marker genes including NDUFA4 (Mitochondrial oxidative phosphorylation), ESRRG (Mitochondrial function), KCNK3 (Katp channel activity) and MAFA (beta cell fate marker). For the human pancreas organoid, amylase secretion and apoptosis of exocrine cells (PI stain) are assayed. For the liver organoid, lipid accumulation is assayed using oil red O or histology. In the case of heart organoid, heart beat and heart size (hypertrophy) are measured. The intestine organoid is analyzed by measuring lipid accumulation using oil red O or histology. 
       FIG. 8A  also shows modeling of human pancreatic cancer tumorigenesis and metastasis in a dish and the potential to screen for drugs that target those diseases. In an exemplary assay, a 3D mini human pancreas is co-cultured with pancreatic cancer cells, stellate cells, and immune cells to create human pancreatic cancer microenvironment in a dish. Various drugs (e.g., FDA-approved drugs) are then screened to find compounds which effectively suppress pancreatic cancer growth or metastasis in a mini human pancreas microenvironment. As output, the following are measured for the pancreas organoid: apoptosis of exocrine cells (PI stain), collagen synthesis (Trichrome stain) and stellate cells activation (GFAP-reporter). Potential candidate drugs identified in these assays are tested in pancreatic cancer tumorigenesis and metastasis mouse models. Genes expression and morphology as well as the degree of cell death, cell growth, and metastasis are investigated. 
       FIG. 8B  provides a schematic showing modeling of human Type 2 diabetes in mice. In an exemplary assay, human islet organoids and/or human liver organoids are transplanted into mice. The mice are then administered various stressors that induce type 2 diabetes, such as a high fat diet (HFD) or cytokines injection. The potential candidate drugs identified in this assay are further tested in human type 2 diabetic mouse model. Genes expression and morphology as well as the degree of diabetes are investigated. 
       FIG. 8B  also shows modeling of human pancreatic cancer tumorigenesis and metastasis in mice. In an exemplary assay, human pancreas organoids and/or human liver organoids are transplanted into mice. Mice transplanted with a mini pancreas are used to study human pancreatic cancer growth in human pancreas microenvironment. In another exemplary assay, a mini pancreas and mini liver are co-transplanted in mice. The liver is a major site for metastasis of pancreatic cancer. In vivo, endothelial cells in the mini pancreas and in the mini liver create a pancreas-liver vasculature network for pancreatic cancer metastasis. Thus, mice co-transplanted with a mini pancreas and mini liver are used to study the metastasis of human pancreatic cancer into the human liver. 
     An ultimate goal of the generation of functional organ-like clusters from pluripotent stem cells (PSC) is to gain insight into the mechanisms underlying human diseases. Although great advances have been made in terms of developing disease models in animals, many of these models fail to faithfully recapture the human condition. In the case of pancreatic islets, their development, cytoarchitecture, and physiology in rodents and human are notably different. 
     Results herein were obtained using the following materials and methods. 
     3D Kelcogel® (3DKG) Culture Media 
     Kelcogel® F low acyl gellan gum (GG-LA) obtained from Modernist Pantry is suspended in pure water 0.3% (w/v) and dissolved by stirring at 90° C. or by microwave. The aqueous solution is sterilized at 121° C. for 20 minutes in an autoclave. The solution is added to TeSR™ (Ludwid et al., Nature methods 3, 637-646) or custom TeSR™ media (800 ml DMEM/F12, 13.28 g BSA. 10 ml Glutamax, 560 mg NaHCO 3 , 330 mg thiamine, 100 mg reduced glutathione, 3300 mg Vitamin C, 14 μg Selenium, 10 ml NEAA, 2 ml Trace element B, 1 ml Trace Element C, 7 μl β-ME, 2 ml DLC, 2 ml GABA, 2 ml LiCl, 129.7 μg pipecolic acid, Insulin 2 mg up to 1000 ml) at final concentration of 0.015%. Methylcellulose (MC) stock solution is added to a final concentration of 0.3% (R&amp;D systems) (e.g., 0.3% Kelcogel® stock: Kelcogel® F low acyl GG-LA 300 mg+MilliQ water 100 ml; 3DKG Stem TeSR™ Base Media: STEMCELL™ TeSR™ 95 ml+0.3% Kelcogel® stock 5 ml+MC stock solution 300 ul; 3DKG Custom TeSR™ Base Media: custom TeSR™ media 95 ml+0.3% Kelcogel® stock 5 ml+MC stock solution 300ul; 1% final concentration of Penicillin/streptozocin is added for 3DKG media). 
     Preparation of Human Pancreatic Endocrine Progenitors and β-Like Cells In Vitro 
     Pancreatic endocrine cells (hiPSC-PEs) were prepared from human iPSC using differentiation methods as previously described. Briefly, human induced pluripotent stem cells (hiPSC) derived from HUVECs were obtained from the Stem Cell Core (Salk Institute). Cells were maintained on MATRIGEL® (BD)-coated dishes in complete STEMCELL™ TeSR™ media at 37 degree in a humidified 5% CO 2  incubator. For pancreatic differentiation, hiPSC were infected with a human insulin reporter lentivirus (pGreenZero lenti reporter human insulin, System Biosciences) by Spinfection (800 g, 1 hour). Methods 1: Media was changed to 100 ng/ml human Activin (R&amp;D Systems), 25 ng/ml recombinant human Wnt3a (R&amp;D Systems) in custom TeSR™ media (800 ml DMEM/F12, 13.28 g BSA, 10 ml Glutamax, 560 mg NaHCO 3 , 330 mg thiamine, 100 mg reduced glutathione, 3300 mg Vitamin C, 14 μg Selenium, 10 ml NEAA, 2 ml Trace Element B, 1 ml Trace Element C, 7 μl (3-ME, 2 ml DLC, 2 ml GABA, 2 ml LiCl, 129.7 μg PA, Insulin 2 mg up to 1000 ml) for 2 days and then 100 ng/ml human Activin in differentiation media for another 2 days (Stage 1, Pancreatic Endoderm). Subsequently, media was replaced with custom TeSR™ media with 1 uM dorsomorphin (Calbiochem), 2 μM Retinoic Acid (Sigma), 10 μM SB431542 and 1% of B27 supplement for 7 days (Stage 2). Media was then replaced with custom TeSR™ media with 10 uM forskolin (Sigma), 10 μM dexamethasone (Stemgent), 10 μM TGFβ RI Kinase inhibitor II/Alk5 inhibitor II (Calbiochem or Enzo), 10 μM Nicotinamide (Sigma), 1 μM 3,3′,5-Triiodo-L-thyronine sodium salt (T3) and 1% of B27 supplement for 4-5 days (day 15-day 21, Pancreatic endocrine progenitors). Media was replaced every day (stage 1) or every other day (stage 2 &amp; stage 3). 
     Methods 2: Media was changed to 100 ng/ml human Activin (R&amp;D Systems), 25 ng/ml recombinant human Wnt3a (R&amp;D Systems) or 3 μM CHIR99021 (Axon or Selleckchem) in differentiation media (51) for 1 day and then 100 ng/ml human Activin in differentiation media (51) for another 2 days (Stage 1, Pancreatic Endoderm). Subsequently, media was replaced with differentiation media (S2) with 50 ng/ml FGF7 (R&amp;D Systems) for 2 days and then differentiation media (S3) with 50 ng/ml FGF7, 0.25 μM SANT-1 (Sigma), 1 μM Retinoic Acid (Sigma), 100 nM LDN193189 and 100 nM α-Amyloid Precursor Protein Modulator TPB for 3 days. Subsequently, media was replaced with differentiation media (S4) with 0.25 μM SANT-1, 50 nM Retinoic Acid, 10 μM Alk5 inhibitor II, 1 μM T3 for 3 days. Subsequently, media was replaced with differentiation media (S5) with 100 nM LDN193189, 100 nM Gamma Secretase inhibitor XX GSiXX (Millipore), 10 μM Alk5 inhibitor II, 1 μM T3 for 7 days. Subsequently, media was replaced with differentiation media (S5) with 10 μM Trolox (Calbiochem), 2 μM R428 (Selleckchem), 1 mM N-acetyl cysteine, 10 μM Alk5 inhibitor II, 1 μM T3 for additional 7 to 20 days. 
     S1 Media (MCDB131 Media, 8 mM glucose, 2.46 g/L NaHCO 3 , 2% Fatty acid free BSA, 0.25 mM L-Ascorbic acid 0.002% Insulin-Transferrin-Selenium ITS-X (GIBCO), 2 mM Glutamax, 1% Penicillin-Streptomycin), S2 Media (MCDB131 Media, 8 mM glucose, 1.23 g/L NaHCO 3 , 2% Fatty acid free BSA, 0.25 mM L-Ascorbic acid, 0.002% Insulin-Transferrin-Selenium ITS-X (GIBCO), 2 mM Glutamax, 1% Penicillin-Streptomycin), S3 Media (MCDB131 Media, 8 mM glucose, 1.23 g/L NaHCO 3 , 2% Fatty acid free BSA, 0.25 mM L-Ascorbic acid, 0.5% Insulin-Transferrin-Selenium ITS-X (GIBCO), 2 mM Glutamax, 1% Penicillin-Streptomycin), S4 Media (MCDB131 Media, 8 mM glucose, 1.23 g/L NaHCO 3 , 2% Fatty acid free BSA, 0.25 mM L-Ascorbic acid, 0.002% Insulin-Transferrin-Selenium ITS-X (GIBCO), 2 mM Glutamax, 1% Penicillin-Streptomycin, 10 μg/ml Heparin, 10 μM Zinc Sulfate), S5 Media (MCDB131 Media or BLAR Media, 20 mM glucose, 1.754 g/L NaHCO 3 , 2% Fatty acid free BSA, 0.25 mM L-Ascorbic acid, 0.002% Insulin-Transferrin-Selenium ITS-X (GIBCO), 2 mM Glutamax, 1% Penicillin-Streptomycin). For 3 dimensional culture, hiPSC or hESC were cultured in 3DKG Stem TeSR™ Base Media with 10 μM Y-27632 for 5 to 7 days and then Media were replaced each Differentiation media with 0.015% Kelcogel and 0.3% Methylcellulose. 
     Generation of Three-Dimensional Pancreatic Islet Bud In Vitro: Islet-Like Organoids in Matrigel Through Co-Culture with hADSCs and HUVECs 
     Primary HUVECs and human Adipose-derived stem cells (hADSC) (Invitrogen or PromoCell) were cultured in 15 cm dish with EBM Medium (Ronza, cc-3121) or MesenProRS™ Medium (GIBCO, 12747-010 or Preadipocyte Growth Medium Kit, C-27417), respectively, at 37 degree Celsius in a humidified 5% CO 2  incubator. For co-culturing experiments, pancreatic endocrine progenitors derived from human iPSC were treated with Accutase, while HUVECs and hADSC were treated with TrypLE (GIBCO, 12604-013) and cells collected into a 50 ml tube respectively. After the cells were counted, 1×10 6  cells of hiPS-PP, 7×10 6  cells of HUVEC and 1-2×10 5  cells of hADSC were co-cultured in 1 well of 24 well with 300 ul of MATRIGEL® matrix. For the purpose of scalable generation of human islets like organoids, 1×10 6  cells of hiPS-PP (day 15-day 21), 7×10 6  cells of HUVEC and 1-2×10 5  cells of hADSC were co-cultured in 3DKG Custom TeSR® media with 10 uM forskolin (Sigma), 10 μM dexamethasone (Stemgent), 10 uM TGFβ RI Kinase inhibitor II/Alk5 inhibitor II (Calbiochem or Enzo), 10 uM Nicotinamide (Sigma), 1 uM 3,3′,5-Triiodo-L-thyronine sodium salt (T3) and 1% of B27 supplement, R428 (204), Zinc sulfate (10 μM) and N-Cys (1 mM). (Methods 1) or co-cultured in differentiation media (S5) with 100 nM LDN193189, 100 nM Gamma Secretase inhibitor XX GSiXX (Millipore), 10 μM Alk5 inhibitor II, 1 μM T3 for 7 days. Subsequently, media was replaced with differentiation media (S5) with 10 μM Trolox (Calbiochem), 2 μM R428 (Selleckchem), 1 mM N-acetyl cysteine, 10 μM Alk5 inhibitor II, 1 μM T3 for additional 7 to 20 days (Methods 2). Mixed cells formed spherical, islet-like clusters within a few days. Media was changed every other day. 
     Generation of 3D (Three-Dimensional) Pancreatic Islet Buds In Vitro: Islet-Like Organoids in Scalable Gellan Gum Through Co-Culture with hADSCs and HUVECs 
     Cells were prepared as described above. Briefly, 1×10 8  cells of hiPS-PP, 2-7×10 7  cells of HUVECs and 5-7×10 6  cells of hADSC were co-cultured in 60-100 ml of 3DKG Custom TeSR™ with 10 μM forskolin (Sigma), 10 μM dexamethasone (Stemgent), 10 μM TGFβ RI Kinase inhibitor II/Alk5 inhibitor II (Calbiochem or Enzo), 10 μM Nicotinamide (Sigma), 1 μM 3,3′,5-Triiodo-L-thyronine sodium salt (T3) and 1% of B27 supplement, R428 (2 μM), Zinc sulfate (10 μM) and N-Cys (1 mM) (Methods 1) or co-cultured in differentiation media (S5) with 100 nM LDN193189, 100 nM Gamma Secretase inhibitor XX GSiXX (Millipore), 10 μM Alk5 inhibitor II, 1 μM T3 for 7 days. Subsequently, media was replaced with differentiation media (S5) with 10 μM Trolox (Calbiochem), 2 μM R428 (Selleckchem), 1 mM N-acetyl cysteine, 10 μM Alk5 inhibitor II, 1 μM T3 for additional 7 to 20 days (Methods 2). Mixed cells formed spherical, islet-like clusters within a few days. Media was changed every day or every other day. 
     Generation of 3D (Three-Dimensional) Pancreatic Islets Bud In Vitro: Islet-Like Organoids in Scalable Gellan Gum 3D Culture Methods without (w/o) Using hADSC and HUVECs 
     Human PSCs, including iPSC or ESC, were initially cultured in matrigel-coated plates (2 dimensional (2D) cultures. Cells were then treated with Accutase (Innovative Cell Technologies, Inc., San Diego, Calif.) to generate a single cell suspension, washed with PBS and centrifuged at 1000-1300 rpm for 5 minutes to pellet cells. Cells were resuspended with 3DKG Stem TeSR™ Base Medium (Stemcell Technologies, Cambridge, Mass.) with 10 μM Y-27632 (a RHO/ROCK pathway inhibitor compound) and cultured for an additional for 5 to 7 days until PSC sphere growth reached 50-100 μm diameter. Media was then replaced with differentiation media supplemented with 0.015% Kelcogel and 0.3% Methylcellulose. The culture medium was changed to differentiation medium (51) containing 100 ng/ml human Activin (R&amp;D Systems), 25 ng/ml recombinant human Wnt3a (R&amp;D Systems) or 3 μM CHIR99021, a glycogen synthase kinase GSK-3 inhibitor (Axon Medchem, Reston, Va.; or Selleckchem) for 1 day and then to differentiation medium (S1) containing 100 ng/ml human Activin for another 2 days (Stage 1, Pancreatic Endoderm). Subsequently, the medium was replaced with differentiation medium (S2) containing 50 ng/ml FGF7 (R&amp;D Systems) for 2 days, and then with differentiation medium (S3) containing 50 ng/ml FGF7, 0.25 μM SANT-1 (Sigma), 1 μM Retinoic Acid (Sigma), 100 nM LDN193189 (an ALK2 and ALK3 inhibitor, Sigma) and 100 nM α-Amyloid Precursor Protein Modulator TPB for 3 days. Subsequently, this medium was replaced with differentiation medium (S4) containing 0.25 μM SANT-1, 50 nM Retinoic Acid, 10 μM Alk5 inhibitor II, 1 μM T3 for 3 days. Subsequently, the medium was replaced with differentiation medium (S5) containing 100 nM LDN193189, 100 nM Gamma Secretase inhibitor XX GSiXX (Millipore) 10 μM Alk5 inhibitor II, 1 μM T3 for 7 days. Subsequently, the medium was replaced with differentiation medium (S5) containing 10 uM Trolox (Calbiochem), 2 μM R428 (Selleckchem), 1 mM N-acetyl cysteine, 10 μM Alk5 inhibitor II, 1 μM T3 for an additional 7 to 20 days. After confirmation of the insulin gene expression by either reporter expression or qPCR (typically on day 20-30), the medium was changed to differentiation medium (S5) containing 10 μM Trolox (Calbiochem), 2 μM R428 (Selleckchem), 1 mM N-acetyl cysteine, 10 μM Alk5 inhibitor II, 1 μM T3 and 100 ng/ml recombinant human (rh)Wnt4 (R&amp;D Systems), 400 ng/ml rhWnt5a, or 50% Wnt5a conditioned medium for 1-20 days. Wnt5a conditioned medium was prepared by culturing an L-Wnt5a cell line (ATCC, CRL-2814) in DMEM with 10% FBS, 1% Penicillin-streptomycin for 4 days after cells had reached 70-100% confluence in T175-T225 Frasko cell culture flasks. 
     Generation of 3D (Three-Dimensional) Liver Bud In Vitro: Organ Buds 
     Hepatocyte cells (hiPSC-HEs) from human iPSC were prepared using differentiation methods as previously described. Briefly, hiPSCs were maintained on MATRIGEL® (BD)-coated dishes in complete STEMCELL™ TeSR™ media at 37 degrees Celsius in a humidified 5% CO 2  incubator. For hepatic differentiation, hiPSC (90% confluence in 6 well) were cultured with 100 ng/ml human Activin (Sigma) and 25 ng/ml recombinant human Wnt3a (R&amp;D systems) or 3 μM CHIR99021 and 1% B27 supplement minus Insulin in RPMI1640 media for 1 day and then 100 ng/ml human Activin and 1% B27 supplement minus Insulin in RPMI media for another 4 days (Stage 1 Hepatic-Endoderm). Subsequently, media was replaced with differentiation media with 10 ng/ml bFGF, 20 ng/ml BMP4 and 1% of B27 supplement in RPMI1640 media for 3 days (Stage 2). Media was then replaced with differentiation media with 0.1 μM Dexamethasone, 20 ng/ml OncostatinM (R&amp;D Systems) and 10-20 ng/ml Hepatic Growth Factor (HGF, R&amp;D Systems) and 1% of B27 supplement in Hepatocyte Culture Media (Lonza, MD, CC-3198, withdraw EGF and Gentamicin/Amphotericin-B) for 4-22 days (day 15-day 19, Pancreatic endocrine progenitors). Media was replaced every day (stage 1) or every other day (stage 2 &amp; stage 3). Primary HUVECs cells and human Adipose-derived stem cells (hADSC) (InVitrogen or PromoCell) were cultured in 15 cm dish with EBM Media (Ronza, cc-3121) or MesenProRS Media (GIBCO, 12747-010 or Preadipocyte Growth Medium Kit, C-27417), respectively, at 37 degree Celsius in a humidified 5% CO 2  incubator. For co-culturing experiments, day 10-hepatocytes derived from human iPSC were treated with Accutase, while HUVECs and hADSC were treated with TrypLE (GIBCO, 12604-013) and cells were collected into 50 ml tube, respectively. After the cells were counted, 1×10 6  cells of hiPS-PP, 7×10 6  cells of HUVEC and 1-2×10 5  cells of hADSC were co-cultured in 1 well of 24 well with 300 ul of matrigel. Liver-like organoids were formed within 1 to 2 days. Then, liver-like organoids were taken out from MATRIGEL® matrix and cultured in in 3DKG Custom TeSR™. 
     Generation of 3D (Three-Dimensional) Heart Bud In Vitro: Organ Buds 
     Cardiomyocyte cells (hiPSC-CDs) were prepared from human iPSC using differentiation methods as previously described. Briefly, hiPSCs were maintained on MATRIGEL® (BD)-coated dishes in complete Stemcell™ TeSR™ media at 37 degree Celsius in a humidified 5% CO 2  incubator. For hepatic differentiation, hiPSC (90% confluence in 6 well) were cultured with 100 ng/ml human Activin (R&amp;D Systems) and 10 uM CHIR99021 and 1% B27 supplement minus Insulin in RPMI1640 media for 1 days and then 1% B27 supplement minus Insulin in RPMI media for another 2 days (Stage 1 cardiac-Mesoderm). Subsequently, media was replaced with RPMI1640 with 5 uM IWP-2 and 1% B27 supplement minus Insulin in RPMI media for 1 days (Stage 2). Media was then replaced with 1% B27 supplement minus Insulin in RPMI Media for 6 days or more (Stage 3). Cardiac contraction started around day 13. Media was replaced every day (stage 1) or every other day (stage 2 &amp; stage 3). Primary HUVECs cells and human Adipose-derived stem cells (hADSC) (Invitrogen or PromoCell) were cultured in 15 cm dish with EBM Media (Ronza, cc-3121) or MesenProRS™ Media (GIBCO, 12747-010 or Preadipocyte Growth Medium Kit, C-27417), respectively at 37 degrees Celsius in a humidified 5% CO 2  incubator. For co-culturing experiments, day 13 to day 15 cardiomyocytes derived from human iPSC were treated with Dispase, while HUVECs and hADSC were treated with TrypLE (GIBCO, 12604-013) and cells collected into 50 ml tube respectively. After the cells were counted, 1×10 6  cells of hiPS-PP, 7×10 6  cells of HUVEC and 1-2×10 5  cells of hADSC were co-cultured in 3DKG Custom TeSR™ media. Mini heart like organs capable of contracting were formed within a few days. 
     Generation of 3D (Three-Dimensional) Intestine Bud In Vitro: Organ Buds 
     Intestinal cells (hiPSC-ITs) were prepared from human iPSC using differentiation methods as previously described. Briefly, hiPSCs were maintained on Matrigel® (BD)-coated dishes in complete Stemcell™ TeSR™ Media at 37 degrees Celsius in a humidified 5% CO 2  incubator. For hepatic differentiation, hiPSC (90% confluence in 6 well) were cultured with 100 ng/ml human Activin (R&amp;D Systems), 3 uM CHIR99021, 2 mM Glutamax and 1% B27 supplement minus Insulin in RPMI1640 media for 1 day and then 100 ng/ml human Activin (R&amp;D Systems), 2 mM Glutamax and 1% B27 supplement minus Insulin in RPMI1640 media for another 3 days (Stage 1 Forgut-Endoderm). Subsequently, media was replaced with 500 ng/ml Wnt3a, 500 ng/ml FGF4 and 1% B27 supplement in RPMI 1640 media for 4 days (Stage 2). Cells were transferred to Matrigel® matrix and then a 3D-spheroid Matrigel® dorm was made in the bottom of 24 well. Media was then replaced with 1% B27 supplement, 1% N2 supplement, 500 ng/ml R-spondin, 100 ng/ml Noggin, 50 ng/ml EGF, 2 mM Glutamax™ supplement, 10 uM HEPES in DMEM/F12 Media for 7 days or more (stage 3). Intestinal-like organoid spheroids were observed within a week. Media was replaced every day (stage 1) and every other day (stage 2 &amp; stage 3). Primary HUVECs cells and human Adipose-derived stem cells (hADSC) (Invitrogen or PromoCell) were cultured in a 15 cm dish with EBM Media (Ronza, cc-3121) or MesenProRS™ Media (GIBCO®, 12747-010 or Preadipocyte Growth Medium Kit, C-27417), respectively, at 37 degrees Celsius in a humidified 5% CO 2  incubator. For co-culturing experiments, intestinal progenitors (day 7) derived from human iPSC were treated with Accutase, while HUVECs and hADSC were treated with TrypLE (GIBCO®, 12604-013) and cells collected into a 50 ml tube respectively. After counting the cells, 1×10 6  cells of hiPS-PP, 7×10 6  HUVEC cells and 1-2×10 5  hADSC cells were co-cultured in 3DKG Custom TeSR™ media. 
     Insulin Secretion Assay (Primary Mouse and Human Pancreatic Islets and Human iPSC-Derived Cells) 
     Insulin release from intact islets was monitored using batch incubation methods (Yoshihara et al., 2010, Nat. Commun. 1:127). Briefly, overnight-cultured isolated pancreatic islets (RPMI-1640 supplemented with 10% (v/v) fetal bovine serum and 1% (v/v) Antibiotic-Antimycotic (Gibco)) were pre-cultured at 37° C. for 30 min (Krebs-Ringer bicarbonate buffer (KRBB) containing 129.4 mM NaCl, 3.7 mM KCl, 2.7 mM CaCl 2 , 1.3 mM KH 2 PO 4 , 1.3 mM MgSO 4 , 24.8 mM NaHCO 3  (equilibrated with 5% CO 2 , 95% O 2 , pH7.4), 10 mM HEPES and 0.2% (v/v) BSA (fraction V, Sigma) (KRBH) with 3 mM glucose). Pancreatic islets were then incubated in KRBH buffer (500 μl/10 islets) with 3 mM or 20 mM glucose to determine insulin secretion levels. After 30 min, islets were pelleted by centrifugation and insulin levels determined by ELISA (Rat/mouse Insulin ELISA KIT (Millipore) and Human Insulin ELISA KIT (Millipore) for mouse and human islets, respectively). For human iPSC derived cells, the cells (1×10 6  cells/well in 24 well) were pre-cultured in 3 mM glucose KRBH buffer (500 μl/well). The cells were then incubated in KRBB (200 μl/well) with 3 mM or 20 mM glucose to determine c-peptide secretion levels as indicator of insulin secretion levels. After 30 min, the cells were pelleted by centrifugation and c-peptide levels were determined by human c-peptide ELISA KIT (Millipore). 
     Quantitative RT-PCR Analysis 
     Total RNA was extracted using TRIzol reagent (Invitrogen) and RNeasy KIT (Qiagen). Reverse transcription was performed with a SuperScript III First-Strand Synthesis System kit (Invitrogen) or PrimeScript RT reagent kit (TAKARA). Real time quantitative RT-PCR (qPCR) was performed using SYBR Green (Bio-Rad). 
     Lentivirus Production for Proinsulin-NanoLuc 
     Proinsulin-NanoLuc in pLX304 (Addgene, #62057) was obtained from Addgene. Proinsulin-NanoLuc lentivirus was produced using a second-generation viral packaging system. Briefly, 14 μg of Proinsulin-NanoLuc, 6.6 μg of PsPAX2 packaging plasmid (Addgene 12260), 5.4 μg of pMD2.G envelope plasmid (Addgene 12259) and 54 μl Lipofectamin2000 (Invitrogen) were used to transfect a T75 flask of HEK293LTV packaging cells. Twenty-four (24) hours after transfection, media was changed to fresh DMEM with 10% FBS and 1% Penicillin/Streptozocine. Forty-eight (48) hours and 96 hours after transfection, viruses were collected as day 1 and day 3, respectively and passed through 0.2 μm cellulose acetate filters (VWR). Viruses were aliquoted and frozen at −80 degrees Celsius until use. 
     Gaussia Luciferase Assay for Insulin Secretion Measurement 
     Mouse islets, human islets and human islets like organoids were plated in their respective growth media with 10 μg/ml Polybrene® polymer (Santacruz). Viruses were then added. After overnight culture, cells were placed in fresh growth media. Forty-eight (48) to 72 hours after infection, mouse islets, human islets and human islet-like organoids were picked up by hand and then placed into 96 wells with single islet or organoid. Then, insulin secretion assays were performed. Briefly, a single islet or organoid was pre-incubated with 3 mM glucose KRBB at 37° C. for 30 min to 1 hour. The cells were then incubated in KRBB (100 μl/well) with 3 mM for 30 min and then sequentially incubated with 20 mM glucose with or without 100 nM Exendin-4 or 3 mM glucose with 20 mM KCl (100 μl/well). To determine Gaussia Luciferase activity as indicator of insulin secretion levels, 10 μl of samples are used for Luciferase assay using Pierce Gaussia Luciferase Flash Assay Kit (Prod #16159, Thermo Scientific). 
     INS-1 cells were infected with the virus by spinfection (800 g, 1 hour at 37 degrees Celsius) and then changed to fresh INS-1 growth media. Seventy-two (72) hours after transfection, INS-1 cells were treated with 5 μg/ml Blasticidin (Invitrogen) for 7 days to select for Proinsulin-NanoLuc expressing cells. For insulin secretion assay, the cells (5×10 4 -1×10 5  cells/well in 96 well) were pre-cultured in 3 mM glucose KRBB (100 μl/well). The cells were then incubated in KRBB (100 μl/well) with 3 mM and then sequentially incubated with 20 mM glucose with or without 100 nM Exendine-4 or 3 mM glucose with 20 mM KCl (100 μl/well). To determine Gaussia Luciferase activity as indicator of insulin secretion levels, 10 μl of samples are used for Luciferase assay using Pierce Gaussia Luciferase Flash Assay Kit (Prod #16159, Thermo Scientific). 
     Vascularization Test In Vitro 
     Human islet-like organoids were embedded in 1 well of 24 well plate with 300 μl of Matrigel® matrix with EBM Media (Ronza, cc-3121). Vascularization was observed within 24-72 hours. 
     3D Culture of hADSCs and WNT Protein Expression 
     hADSCs undergo changes in the expression of Wnt genes, in particular genes in the Wnt5a pathway, during the spontaneous self-organization that occurs in 3D culture. ( FIG. 11A ). Wnt5a was found to be the predominant protein expressed among the Wnt proteins in hADSC 3D culture over time. ( FIG. 11B ). 
     Example 5: Wnt Proteins in the Metabolic Maturation of iPSC-Derived Islet Organoids 
     Fltp and Esrrg genes were found to be expressed in iPSC-derived islet organoids (day 21, generated without co-culture with hADSCs or HUVECs) after treatment with PBS, WNT3a (500 ng/ml), recombinant human (rh)WNT4 (100 ng/ml), or rhWNT5a (400 ng/ml) for 5 days. ( FIG. 12A ). As shown in  FIG. 12B , Esrrg gene expression was induced in hiPSC-derived islet organoids that were generated in the absence of supporting hADSC or HUVECs, in response to increasing doses of rhWNT4 (0, 10, 25, 50, 100, 200 ng/ml) and rhWNT5a (0, 25, 50, 100, 200, 400 ng/ml). In addition, mitochondrial genes involved in oxidative phosphorylation (Cox7a2, Ndufa1, Ndufa7), lactate dehydrogenase (Ldha) and Fltp (a Wnt/planar cell polarity (PCP) effector and reporter gene) were induced in hiPSC-derived islet organoids that were generated in the absence of supporting hADSC or HUVECs, in response to increasing doses of rhWNT4 (0, 10, 25, 50, 100, 200 ng/ml) and rhWNT5a (0, 25, 50, 100, 200, 400 ng/ml), ( FIG. 12C ). Mitochondrial (Mitotracker; Mito-Red) and insulin (Insulin-GFP) levels were increased in hiPSC-derived islet organoids (day 27) after 8 days treatment with PBS or WNT4 (100 ng/ml). ( FIG. 12D ). Human iPSC-derived islet organoids (day 27) were generated after 8 days treatment with PBS or WNT4 (100 ng/ml). ( FIG. 12E ). Insulin production was found in hiPSC-derived islet organoids (day 27) after 8 days treatment with rhWNT4 (100 ng/ml), rhWNT5a (400 ng/ml), or WNT5a secreting fibroblast conditioned media (50%), compared with PBS and control fibroblast conditioned media (50%). ( FIGS. 12F-12H ). Human iPSC (hiPSC)-derived islet organoids (day 22) treated with rhWnt4 (100 ng/ml) for 12 days showed functional maturation based on their secretion of human c-peptide, as measured in response to low glucose βmM, “G3 mM”), high glucose (20 mM, “G20 mM”), or high KCl levels (20 mM, “KCL20 mM”), ( FIG. 13 ). 
     OTHER EMBODIMENTS 
     From the foregoing description, it will be apparent that variations and modifications may be made to the invention described herein to adopt it to various usages and conditions. Such embodiments are also within the scope of the following claims. 
     The recitation of a listing of elements in any definition of a variable herein includes definitions of that variable as any single element or combination (or subcombination) of listed elements. The recitation of an embodiment herein includes that embodiment as any single embodiment or in combination with any other embodiments or portions thereof. 
     All patents and publications mentioned in this specification are herein incorporated by reference to the same extent as if each independent patent and publication was specifically and individually indicated to be incorporated by reference.