tbuckley/cabot-search huggingface dataset:
393. Effectiveness of Antioxidant Gene Therapy in Delaying Retinal Degeneration
Authors: Manas R. Biswal, Cristhian Ildefonso, Zhaoyang Wang, Haoyu Mao, Hong Li, Ping Zhu, Giovani Quinones, Alfred S. Lewin
Journal: Molecular Therapy (2015)

We established a mouse model of RPE(retinal pigment epithelium) oxidative stress by Cre-lox mediated deletion of the Sod2 gene, that codes for the protective enzyme manganese superoxide dismutase (MnSOD), leading to some of the features of geographic atrophy. It is the purpose of these experiments to determine whether early delivery of Sod2 using adeno-associated virus (AAV) can prevent retinal degeneration seen in these mice and whether gene therapy can prevent degeneration once it has begun. Deletion of Sod2 was induced by doxycycline treatment of mice with a "floxed" allele of Sod2 and an RPE-specific tet-transactivator controlling expression of Cre (Sod2-cre mice). Retinal degeneration was monitored by electroretinography (ERG) and spectral domain optical coherence tomography over a period of 9 months. Mouse Sod2 with a Myc epitope under the control of a small chicken beta- actin promoter (smCBA) was packaged into self-complementary AAV serotype 1 vector (ScAAV1). Ten Sod2-cre adult mice were injected subretinally with ScAAV1-smCBA-Sod2-Myc or ScAAV1-smCBA-GFP into right and left eye respectively. Three mice were sacrificed a month following injection to collect retina and RPE separately. Using an anti-myc antibody, western blotting was performed to detect MnSOD expression. For rest of the mice ERG and SD-OCT were measured at different time points. Following doxycycline induction of Cre, mice demonstrated increased signs of oxidative stress in RPE and accumulation of autofluorescent material by 2 months of age. They showed a gradual decline in the ERG response and thinning of the outer nuclear layer (by SD-OCT) which were statistically significant by 6 months. Myc tagged MnSOD expression was detected in RPE of mice injected with vector driven by smCBA promoter and negligible expression was seen in the neural retina. ERG and OCT data suggests no adverse functional and structural integrity due to increased expression of MnSOD and sub retinal injection. ERG response and thinning retinal thickness was significantly delayed in Sod2-vector injected eyes compared to control eye injected with GFP vector. Deletion of Sod2 in the RPE leads to some of the salient features of dry AMD. ScAAV1 delivery of Sod2 led to expression in RPE. Delivery of ScAAV1-smCBA-SOD2-Myc vector can be used as a tool to reverse oxidative stress in this mouse model of dry AMD.
756. Stable Amelioration of Hemophilia B in Dogs by Intravenous Administration of Lentiviral Vectors Expressing Hyper-Functional Factor IX
Authors: Alessio Cantore, Michela Milani, Tongyao Liu, Patrizia Della Valle, Armando D'Angelo, Thierry VandenDriessche, Marinee Chuah, Haiyan Jiang, Timothy Nichols, Luigi Naldini
Journal: Molecular Therapy (2016)

Lentiviral vectors (LVs) are attractive vehicles for liver-directed gene therapy by virtue of their ability to stably integrate in the genome of target cells and the low prevalence of pre-existing immunity against HIV in humans. Over the past years, we have developed a LV platform that can achieve stable transgene expression in the liver, induce transgene-specific immune tolerance and establish correction of hemophilia in mouse models upon systemic administration. This LV is designed to stringently target transgene expression to hepatocytes through transcriptional and microRNA-mediated regulation. We then investigated the efficacy and safety profile of portal vein administration of LVs expressing canine factor IX (FIX) in a canine model of hemophilia B. We produced large-scale batches of LVs qualified for in vivo administration and treated adult hemophilia B dog by portal vein administration. We observed long-term stable reconstitution of canine FIX activity up to 1% of normal and significant amelioration of the clinical phenotype in 3 treated dogs with 6, 3.5 and 2.5 years of follow up. LV infusion was associated with transient signs of inflammatory response and mild hepatotoxicity, which could be abrogated by pretreatment with anti-inflammatory drugs. There was no detectable long-term toxicity or development of FIX inhibitors. In the perspective of clinical translation and to increase therapeutic efficacy, we next treated two 10-kg hemophilia B dogs by peripheral vein administration of LVs expressing a codon-optimized and hyperfunctional canine FIX at a 5-fold higher dose than those previously administered. Intravenous LV administration was well tolerated with mild and self-limiting elevation of aminotransferases in one dog. In the dog that reached more than 1 year of follow up FIX activity ranged between 4-8% of normal. Treatment of two more dogs at a higher dose is underway. Overall, our studies position LV-mediated liver gene therapy for further pre-clinical development and clinical translation. LVs may thus complement other available vectors to address some of the outstanding challenges posed by liver gene therapy of hemophilia and conceivably other diseases.
Abstract 5569: p73 dinucleotide polymorphism genotyping of human cancer cell lines
Authors: L. Michael Carastro, Kaia K. Hampton, Ricardo A. Cordova Estupinan, Hyun Y. Park, Dongwha Kim, Jong Y. Park
Journal: Cancer Research (2015)

Abstract The p73 gene is a member of the p53 tumor suppressor family. The p73 dinucleotide polymorphism (DNP) (rs1801173) is a G4C14-to-A4T14 linked pair of transitions located in exon 2 between the P1 and P2 gene promoters. The P1 and P2 p73 gene promoters are the transcription initiation sites for mRNAs encoding TAp73 and ΔNp73 isoforms, respectively. Recently, we reported the p73 DNP allele was associated with (1) decreased risk [OR = 0.55, 95%CI = 0.31-0.99] for aggressive prostate cancer (PCa) and (2) increased TAp73/ΔNp73 protein isoform ratios in ten human cancer cell lines. We hypothesize the presence of the p73 DNP allele causes altered p73 promoter usage resulting in increased TAp73/ΔNp73 protein isoform ratios, which could explain the observed decreased risk for PCa aggressiveness. Our ultimate goal is to assess the potential of p73 DNP as a biomarker for lower aggressive PCa risk. Therefore, our initial aim in this study was to determine the p73 DNP genotype in ten additional human cancer cell lines (DU 145, JEG-3, K-562, LNCaP, MDA-MB-231, MDA-MB-468, MRC-5, PC-3, SW48 and U-2OS) not yet characterized for p73 DNP status. We previously reported the p73 DNP genotype of three human cancer cell lines that we used as genotyping controls: Caco-2 (homozygous polymorphic), NCI-H1299 (homozygous wild type), and HepG2 (heterozygous). Cell lines were cultured and genomic DNA was isolated and quantitated. Cell line genomic DNA samples were used in TaqMan Real Time-PCR allelic discrimination assays to determine the p73 DNP genotypes. Our data conclusively determined that DU 145, JEG-3, K-562, LNCaP, MDA-MB-468, MRC-5, SW48 and U-2OS were p73 DNP homozygous wild type, while PC-3 and MDA-MB-231 werw p73 DNP heterozygous. Cell Line p73 DNP GenotypesCELL LINE NAMEP73 DNP GENOTYPEDU145CCJEG-3CCK-562CCLNCaPCCMDA-MB-231CTMDA-MB-468CCMRC-5CCPC-3CTSW48CCU-2OSCC Citation Format: L. Michael Carastro, Kaia K. Hampton, Ricardo A. Cordova Estupinan, Hyun Y. Park, Dongwha Kim, Jong Y. Park. p73 dinucleotide polymorphism genotyping of human cancer cell lines. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5569. doi:10.1158/1538-7445.AM2015-5569
Abstract 3836: Identification of key survival-correlating microRNAs and Piwi-interacting RNAs dysregulated in head and neck squamous cell carcinoma
Authors: Jonjei Ku, Angela E. Zou, Thomas K. Honda, Hao Zheng, Maarouf A. Saad, Vicky Yu, Yinan Xuan, Pranav Singh, Mehran Rahimy, Selena Z. Kuo, Weg M. Ongkeko, Jessica Wang-Rodriguez
Journal: Cancer Research (2015)

Abstract Aberrant expression of microRNAs and piwi-interacting RNAs, two distinct classes of non-coding RNA (ncRNA) and important regulators of gene expression, is known to be implicated in human diseases including cancer. The exact roles of microRNAs and piwi-interacting RNAs in the pathogenesis of head and neck squamous cell carcinoma (HNSCC), however, remain poorly understood. Utilizing next generation RNA sequencing analysis of raw data obtained from The Cancer Genome Atlas, we discovered significant dysregulation (fold change ≥ 2) in 231 microRNAs and 60 piwi-interacting RNAs between forty-three matched samples of tumor and adjacent normal tissues, including down-regulation of mirs 30e and let7-c, and up-regulation of piR34736 and mir-196a-1 in tumor tissue compared to adjacent, non-tumor tissue. Further statistical survival modeling using patient survival data and cox proportional hazard regression analysis revealed that miRs 30e, 196a-1, and let7-c, and piR34736 correlated significantly with survival in patients with HNSCC. To further investigate these microRNAs and piwi-interacting RNAs, quantitative reverse transcriptase PCR (qRT-PCR) and MTT cell proliferation assays were performed in-vitro to assess their isolated effects on cancer gene expression in established HNSCC cell lines. Endogenous expression of these ncRNA was compared between a set of three normal oral keratinocytes and five HNSCC cell lines using qRT-PCR which confirmed miR-30e and mir-let7-c down-regulation and piR34736 and mir-196a-1 up-regulation in cancer. MTT cell proliferation assays upon knock-down of miR-196a-1 and ectopically expressed mir-let7-c in HNSCC revealed significant decreases in cell proliferation compared to cells transfected with their respective negative control. Identification of key ncRNAs differentially expressed between cancer tissue and adjacent normal tissue may reveal important steps in the initiation and progression of HNSCC as they are strongly correlated with patient survival. These key microRNAs and piwi-interacting RNAs may serve as potential biomarkers and therapeutic targets for future epigenetic-based cancer therapies. Citation Format: Jonjei Ku, Angela E. Zou, Thomas K. Honda, Hao Zheng, Maarouf A. Saad, Vicky Yu, Yinan Xuan, Pranav Singh, Mehran Rahimy, Selena Z. Kuo, Weg M. Ongkeko, Jessica Wang-Rodriguez. Identification of key survival-correlating microRNAs and Piwi-interacting RNAs dysregulated in head and neck squamous cell carcinoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3836. doi:10.1158/1538-7445.AM2015-3836
Bioreactors for haematopoietic cell culture
Authors: L K Nielsen
Journal: Annual Review of Biomedical Engineering (1999)

Hematopoietic cell culture, or ex vivo expansion of hematopoietic cells, is an enabling technology with many potential applications in bone-marrow transplantation, immunotherapy, gene therapy, and the production of blood products. Hematopoietic cultures are complex, with many different cell types at different stages of development present at any given point in time and never in steady state. Moreover, these cells interact strongly with each other and the environment through cytokines (growth factors) and adhesion molecules, as well as through their metabolism. Despite these significant challenges, cell products produced in bioreactors have shown promise in recent phase 1 clinical trials. Copyright
Abstract IA06: Serum contains enough information for early detection of pancreatic cancer
Authors: Carl A. K. Borrebaeck
Journal: Cancer Research (2016)

Abstract One of the driving forces in personalized medicine is the search for protein patterns that can be utilized for improved diagnosis, assessing the risk for tumor relapse and disease progression. Affinity proteomics, a high-throughput technology for protein expression profiling, is now generating data for improved diagnosis and patient stratification, which has been demonstrated in several clinical studies. Pancreatic ductal adenocarcinoma (PDAC) is such a disease where detection preceding clinical symptoms significantly increases the life expectancy of patients. Based on multiparametric microarray analysis of over 2500 patient samples, serum protein signatures have for the first time been identified that could discriminate between pancreatic cancer patients with stage I, II, III and IV, which indicates a possibility for early diagnosis and an increased detection rate of surgically resectable tumors, which significantly would increase the life expectancy of pancreatic cancer patients. Citation Format: Carl A. K. Borrebaeck.{Authors}. Serum contains enough information for early detection of pancreatic cancer. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2016 May 12-15; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2016;76(24 Suppl):Abstract nr IA06.
Alopecia areata incognita in Cronkhite-Canada syndrome
Authors: S. Ong, C. Rodriguez-Garcia, S. Grabczynska, J. Carton, M. Osborn, J. Walters, F. Kubba, C.M. Stefanato
Journal: British Journal of Dermatology (2016)

Cronkhite–Canada syndrome is an acquired inflammatory polyposis syndrome in which alopecia, onychomadesis and hyperpigmentation occur concurrently with gastrointestinal symptoms. The pathophysiology of alopecia in Cronkhite–Canada syndrome has not been definitively elucidated. We present evidence for alopecia areata incognita as a possible mechanism of hair loss.
Model-Based Characterization of the Molecular Response Dynamics of Tyrosine Kinase Inhibitor (TKI)-Treated CML Patients – a Comparison of Imatinib and Dasatinib First-Line Therapy
Authors: Ingmar Glauche, Christoph Baldow, Sabine Fröhlich, Philipp Schulze, Amit Roy, Milayna Subar, Xiaoning Wang, Ingo Roeder
Journal: Blood (2014)
321. Deletion of Mutated GAA Repeats from the Intron 1 of the Frataxin Gene Using the CRISPR System Restores the Protein Expression in a Friedreich Ataxia Model
Authors: Dominique L. Ouellet, Joel Rousseau, Khadija Cherif, Catherine Gérard, Renald Gilbert, Alain Garnier, Bruno Gaillet, Christian Beauséjour, Jacques P. Tremblay
Journal: Molecular Therapy (2016)

The CRISPR system is now widely used as a molecular tool to edit the genome. We used this technique in Friedreich Ataxia (FRDA), an inherited autosomal disease known to cause a decrease of the mitochondrial frataxin protein. Genetic analysis revealed a GAA repeat expansion within the intron 1 of the frataxin (FXN) gene. We used cells derived from the YG8sR mouse model where the mouse frataxin gene is knockout but contain a human FXN mutated transgene on one allele. We then deleted the GAA trinucleotide repeat using 2 specific guide RNAs (gRNAs) co-expressed with either S. pyogenes (Sp) or S. aureus (Sa) Cas9. We were able to monitored an increase up to 2-fold of frataxin mRNA and protein levels in clone cells. We also confirmed these results in vivo using DNA electroporation in the Tibialis anterior muscle of the YG8R mice. Ongoing in vivo investigation of a systemically injected AAV-DJ vector expressing the SaCas9 and 2 successful selected gRNAs in the mouse model YG8sR will hopefully provide more details answers on the efficacy of the approach and give us preliminary data to go forward for clinical trial. The deletion of the GAA repeats expansion then might be a highly valuable gene therapy approach for FRDA patients.
591. MiRNAs Regulates PVRL4 and Oncolytic Measles Virus Infectivity in Glioblastoma and Breast Cancer
Authors: Hirosha Geekiyanage, Mark Jentoft, James Ingle, Evanthia Galanis
Journal: Molecular Therapy (2015)

Oncolytic measles virus strains are currently being evaluated in several clinical trials including glioblastoma and ovarian cancer, while clinical application in breast cancer is in development. The role of poliovirus receptor-related 4 (PVRL4), as a measles virus receptor, has been well documented. However the regulation of PVRL4 is not well understood. Immunohistochemistry and western blot analysis shows increased PVRL4 protein levels in glioblastoma and breast tumor clinical samples with no significant change in PVRL4 mRNA levels detected by qRT-PCR, suggesting that PVRL4 is likely post-transcriptionally regulated. Genes can be post-transcriptionally regulated by miRNAs by binding to the 3'UTR of the targeted mRNA causing translational repression or degradation. Thus, we sought to investigate the potential role of miRNAs in PVRL4 post-transcriptional regulation. Algorithms from microRNA. org and RegRNA, predicted that, from the miRNAs that are reportedly down-regulated in glioblastoma and breast cancer, miR-31 and miR-128 binds the 3'UTR of PVRL4. Indeed, luciferase binding assays demonstrated miR-31 and-128binding to the 3'UTR of PVRL4, suggesting that PVRL4 is a miRNA targeted gene. QRT-PCR studies validated that miR-31 and miR-128 are down-regulated in glioblastoma and breast tumor clinical samples, further supporting the hypothesis that PVRL4 are post-transcriptionally regulated by miRNAs. In order to understand the effects of the respective miRNAs on PVRL4 and thus on measles virus infectivity, a viral strain (Leonard et. al, 2010, J. Virol.) that exclusively utilizes PVRL4 as its receptor was employed to infect the cancer cell lines followed by evaluation of virus infectivity via measles viral nucleoprotein (N-protein) levels. Subsequent gain -and loss-of-function experiments showed that overexpression of miR-31and miR-128, via lentiviral-vector transduction, down-regulated the endogenous levels of PVRL4 and measles virus infectivity, while transduction of lentiviral-vectors expressing PVRL4 restored measles virus infectivity. The functional affects were reversed or increased upon transduction with the complementary anti-miRs-128 and anti-miR-31. These results are being further validated in ongoing in vivo mouse models where cancer cell lines stably transduced with miR-128, miR-31 or anti-miR-128, anti-miR-31, are transplanted in mice to evaluate tumor size and survival rates upon measles virus infection. These data suggest that PVRL4 is post-transcriptionally regulated by miR-128 and miR-31, and thus, these represent possible miRNA targets that could modulate PVRL4 levels in order to enhance oncolytic measles virus infectivity. This research is supported by P50 CA 108961-7, P50 CA 136393-4, P50 CA 116201-7 and R01 CA 136547-4.
359. Targeted Genome Editing Using TALENs to Correct a Mouse Model of Methylmalonic Acidemia (MMA)
Authors: Jessica L. Schneller, Randy J. Chandler, Charles P. Venditti
Journal: Molecular Therapy (2016)

Methylmalonic acidemia (MMA) is an autosomal recessive metabolic disorder, in which the body is unable to oxidize valine and isoleucine as well as odd chain fatty acids. Approximately 60 percent of MMA cases are caused by mutations in methylmalonyl-CoA mutase (MUT), the enzyme that catalyzes the isomerization of methylmalonyl-CoA to succinyl-CoA. Current treatments for MUT class MMA include dietary protein restriction and cofactor supplementation; however patients continue to exhibit severe morbidity and high rates of mortality. A murine model of MMA was created by knocking out exon 3 of the Mut gene and exhibits the severest clinical phenotype of MMA, neonatal lethality. Gene therapy treatments using recombinant adeno-associated viral vectors to treat mice with MMA have proven effective, however long-term follow-up of mice treated as neonates revealed genotoxicity caused by AAV integrations, leading the mice to develop hepatocellular carcinoma. Programmable nucleases such as TALENs should make it possible to correct at the MUT locus, ameliorate disease symptoms and reduce the effects of off-target integration by precise genome modification. Furthermore, AAV-mediated delivery of TALENs with an HR cassette carrying a rescue cDNA for the Mut gene in a disease model of MMA should enable in vivo correction in the liver, and ameliorate the MMA phenotype. We have designed TALENs that target intron 2 of the Mut gene and a partial rescue cassette that contains codon-optimized exons 3 to 13 of the Mut gene to restore Mut expression in the exon 3 knock out model of MMA after successful homologous recombination. The TALENs are expressed under the control of the liver-specific thyroxine binding globulin promoter to ensure cleavage in hepatocytes, and delivered using a hepatotropic AAV serotype 8 capsid. To restore expression and achieve targeted integration, the rescue cassette is preceded by a splice acceptor site and flanked by homology arms around the TALEN cleavage site. Integration of the rescue cassette will restore Mut expression at the locus, using the endogenous promoter and exons 1 and 2 of the Mut gene to reconstitute a functional Mut gene. AAV packaging limitations necessitate two AAV vectors for delivery of the TALENs, which are 4kb per pair, and required regulatory elements, as well as a third vector for delivery of the Mut homologous recombination rescue cassette. All three vectors will be delivered systemically in the neonatal period. We hypothesize that in vivo editing at the Mut locus with subsequent HR will provide sufficient Mut enzyme activity to improve the clinical and biochemical phenotypes of the MMA mice, with minimal off-target integration, thereby decreasing the risk of genotoxicity associated with AAV gene therapy.
Epigenetic Silencing of BIM Mediates Chemotherapy Resistance of Patients with Burkitt Lymphoma That Can Be Overcome by Therapeutic Reactivation of BIM in Mouse and Human Lymphoma Models
Authors: Jose A. Richter, Antonio Rullan, Elena Beltran, Xabier Agirre, M.a José Calasanz, Jose Roman-Gomez, Carlos Panizo, Jesus M Hernandez, Felipe Prosper, Vicente Fresquet, Jose A. Martinez-Climent
Journal: Blood (2008)
Abstract 1043: Blocking constitutive aryl hydrocarbon receptor (AhR) signaling attenuates androgen receptor activity
Authors: Maryam Ghotbaddini, Aaron Hemphill, Joann B. Powell
Journal: Cancer Research (2015)

Abstract PURPOSE: The aryl hydrocarbon receptor (AhR) is a member of the basic-helix-loop-helix family of transcription factors. AhR is widely known for regulating the transcription of drug metabolizing enzymes involved in the xenobiotic metabolism of carcinogens and therapeutic agents, such as cytochrome P450-1B1 (CYP1B1). Additionally, AhR has also been reported to interact with multiple signaling pathways during prostate development. Here we investigate the effect of sustained AhR signaling on androgen receptor function in prostate cancer cells. METHODS/RESULTS: Immunoblot analysis shows that AhR expression is increased in androgen independent (C4-2) prostate cancer cells when compared to androgen sensitive (LNCaP) cells. qRT-PCR studies revealed constitutive AhR signaling in C4-2 cells without the ligand induced activation required in LNCaP cells. A reduction of AhR activity by short RNA mediated silencing in C4-2 cells reduced expression of both AhR and androgen responsive genes. The decrease in androgen responsive genes correlates to a decrease in phosphorylated androgen receptor and androgen receptor expression in the nucleus. CONCLUSION: These data indicates that AhR is required to maintain hormone independent signaling by the androgen receptor in C4-2 cells. Collectively, these data provide evidence of a direct role for AhR in androgen independent signaling and provides insight into the molecular mechanisms responsible for sustained androgen receptor signaling in hormone refractory prostate cancer. These studies were supported by the RCMI grant 2G12RR003062-22. Citation Format: Maryam Ghotbaddini, Aaron Hemphill, Joann B. Powell. Blocking constitutive aryl hydrocarbon receptor (AhR) signaling attenuates androgen receptor activity. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1043. doi:10.1158/1538-7445.AM2015-1043