Patent Publication Number: US-2022226384-A1

Title: Extracellular vesicle compositions and methods of use thereof

Description:
CROSS-REFERENCE TO RELATED APPLICATION 
     This application claims the benefit of and priority to U.S. Ser. No. 63/138,969 filed Jan. 19, 2021, and U.S. Ser. No. 63/212,987 filed Jun. 21, 2021, each of which is incorporated by referenced herein in its entirety. 
    
    
     REFERENCE TO THE SEQUENCE LISTING 
     The Sequence Listing submitted as a text file named “EVIA_100_ST25” created on Jan. 18, 2022 and having a size of 15,353 bytes is hereby incorporated by reference pursuant to 37 C.F.R. § 1.52(e)(5). 
     FIELD OF THE INVENTION 
     The field of the invention generally relates to cell-free compositions including extracellular vesicles and methods of use thereof. 
     BACKGROUND OF THE INVENTION 
     Lymphedema is based on a chronic disorder of the lymphatic system and accumulation of interstitial protein-rich fluid in the limbs. Lymphedema patients demonstrate chronic inflammation, and disorders of sensory and motor systems. Generally, lymphedema can be divided into primary and secondary types depending on the etiology. Primary lymphedema results from anatomic or functional defects, whereas secondary lymphedema is due to mainly infection or surgical resection of the lymph node for cancer therapy. It is estimated that approximately 30% of patients who undergo breast cancer surgery may develop lymphedema and 6% of cases of sentinel navigation surgery in the breast progress to lymphedema (DiSipio et al., Lancet Once, 14, 500-515 (2013)). In gynecologic cancers, approximately 10 to 30% of patients may develop lymphedema, depending on the cancer type, age, surgical approach. Furthermore, postoperative radiation increases the proportion of lymphedema to over 35% (Beesley et al.,  Cancer,  109, 2607-2614 (2007), Tada et al.,  BMC Cancer,  9, 47 (2009)). Because lymphedema disturbs the quality of life and elevates risks of recurrent cellulitis and secondary malignancy, the urgent development of effective treatment options is an important clinical goal. 
     To date, the treatment options are mainly limited to physiotherapy and surgical treatments. These methods require proper compliance and lifelong care. As the conventional treatment option, compression therapy using bandages or stockings is still used all over the world. Compression therapy can increase the internal pressure of the lymphatic systems. Thus, interstitial fluid could go into the lymphatic systems. Ideally, patients are required to wear thick garments or stockings every day even in summer; however, for most of the patients, full compliance is hard to achieve. In recent years, with the development of techniques using microsurgery and supermicrosurgery, surgical interventions such as lymphatico-venous anastomosis (LVA) have been attempted worldwide, with consistently favorable results. The long-term outcome of lymphatic microsurgery is also reported to be favorable in 24 to 48 months (Pedro, et al.,  Microsurgery,  40(2):130-136 (2020), and Antonio, et al.,  Gland Surg.,  9(2):539-544 (2020)). However, these techniques are not easy to perform and are therefore not widely used in conventional clinical settings. Furthermore, it is still controversial whether the effect of surgical interventions lasts for an extended period. Moreover, such interventions cannot always stop the progress of the disease in severe cases. Regeneration of the lymphatic system is a hopeful treatment for the disease. Recent reports indicate that transplantation of mesenchymal stem cells (MSCs), such as adipose-derived mesenchymal stem cells (ADSCs) or bone-marrow mesenchymal stem cells (BMMSCs), promotes tissue regeneration including the lymphatic system by paracrine factors secreted by MSCs (Maertens et al.,  PLoS One,  September 15;9(9):e106976 (2014), Shimizu et al.,  J Am Heart Assoc.,  August;1(4):e000877 (2012), Spees, et al.,  Stem Cell Res Ther.,  (7:125 (2016)). Diseases involving vascular and lymphatic systems such as ischemic heart failure, ischemic limb, DM foot necrosis, and lymphedema are possible disease indications. Transplantation of MSCs into damaged tissue has been shown to induce endothelial cell growth and enhance new blood vessel formation, with secreting paracrine factors as the predominant mechanism (Zhang, et al.,  J Transl Med.  (13:49) (2015)). 
     As for inducing lymphangiogenesis, MSCs including ADSCs or BMMSCs have been shown to secrete many growth factors and cytokines that have effects on cells in their vicinity (Hwang, et al.,  Biomaterials,  (32(19):4415-23 (2011)). For example, ADSCs can restore the lymphatic vascular network in a secondary mouse lymphedema model with increased collecting lymphatic vessels, mainly based on paracrine effects of ADSCs (Yoshida et al.,  Regen Med.,  10(5):549-62 (2015)). In addition, it has been shown that BMMSCs play a role in lymphatic regeneration in a mouse tail lymphedema model (Conrad et al.,  Circulation, January  20;119(2):281-9 (2009)). In a clinical trial, BMMSCs were used for 10 post-mastectomy lymphedema patients, and the results indicate that BMMSC injection reduces arm volume as well as associated co-morbidities of pain (Maldonado et al.,  Cytotherapy, November; 13(10):1249-55 (2011), Maldonado, et al.,  Cytotherapy,  (13(10):1249-55 (2011)). ADSCs have been also used in both animal and clinical trials, showing their lymphangiogenic activity and therapeutic efficacy without serious adverse events in the six months follow-up period (Toyserkani et al.,  Stem Cells Transl Med.,  August;6(8):1666-1672 (2017), Hwang et al.,  Biomaterials,  July;32(19):4415-23 (2011)). Potential risks of previous cancer spreading after treatment using MSCs are thought to be low because lymphedema treatment normally starts several years after initial treatment. However, MSC transplantation has drawbacks such as poor engraftment efficiency, potential tumor formation, unwanted immune responses, non-specific differentiation, and the difficulty of quality control before administration (Zhang, et al.,  Cell Pro  49:3-13 (2016)). Thus, there remains a need for alternatives to cell transplantation. 
     Thus, it is an object of the invention to provide alternative compositions and methods to mesenchymal stem cell transplantation. 
     It is a further object of the invention to provide compositions and methods of use thereof to promote generation or regeneration of the lymphatic system, treat lymphedema, and combinations thereof. 
     SUMMARY OF THE INVENTION 
     Compositions and methods for promoting generation or regeneration of the lymphatic system in a subject are provided. Compositions and methods for the treatment of lymphedema are also provided. The compositions include extracellular vesicles, typically in a pharmaceutically acceptable carrier, and are typically cell-free. In some embodiments, the extracellular vesicles are formed by a method including culturing MSCs to produce media conditioned with the extracellular vesicles, and optionally, but preferably separating the extracellular vesicles from the conditioned media. Thus, in some embodiments, the composition does not include the media conditioned by the MSCs. The MSCs can be primary cells or a cell line. MSCs can be isolated or derived from, for example, bone barrow, placenta, umbilical cord blood, adipose tissue, adult muscle, corneal stroma, or the dental pulp of deciduous teeth. In a preferred embodiment, the composition includes extracellular vesicles formed by adipose-derived stem cells. 
     The extracellular vesicles can include or consist of ectosomes, microvesicles (MV), microparticles, exosomes, oncosomes, apoptotic bodies (AB), tunneling nanotubes (TNT), or a combination thereof. In some embodiments, the extracellular vesicles include or consist of exosomes, microvesicles or a combination thereof. The extracellular vesicles can include or consist of vesicles having a size of, for example, between about 20 nm and about 500 nm, or between about 20 nm and about 250 nm, or between about 20 nm and about 200 nm, or between about 20 nm and about 150 nm, or between about 20 nm and about 100 nm. In some embodiments, extracellular vesicles of the composition include CD9, CD63, or a combination thereof. In some embodiments, extracellular vesicles of the composition include one or more of miR-199a-3p, miR-145-5p, miR-143-3p, miR-377-3p, miR-100-3p, miR-29a-3p, miR-495-3p, miR-29c-3p, miR-658, miR-493-3p, miR-184, and miR-27a-3p. 
     In some embodiments the composition can increase the proliferation, migration, and/or tube formation of lymphatic endothelial cells, increase expression of one or more lymphatic markers (e.g., hyaluronan receptor-1 (LYVE-1), vascular endothelial growth factor receptor-3 (VEGFR-3), prospero homeobox 1 (Prox1), and/or podoplanin) in lymphatic endothelial cells, increase angiogenesis, increase lymphangiogeneisis, reduce inflammatory response, decrease fibrosis formation, enlarge circumference and/or induce formation of capillary vessels and/or lymphatic vessels, induce formation of vessels that express both vascular and lymphatic markers, increase drainage routes (e.g., for accumulated fluids), increase HIF1-alpha expression and/or activity (e.g., in lymphatic endothelial cells), reduce Prohibitin (PHB) expression and/or activity (e.g., in lymphatic endothelial cells), or a combination thereof in the subject. 
     The methods typically include administering a subject in need thereof a composition having an effective amount of extracellular vesicles formed by mesenchymal stem cells (MSCs) to, for example, increase generation of the lymphatic system or reduce one or more symptoms of lymphedema. The subject can have lymphedema or one or more symptoms thereof such as swelling of part or all of the arm(s) and/or leg(s), a feeling of heaviness or tightness, restricted range of motion, aching or discomfort, recurring infections, and fibrosis in one or both arms and/or legs. The subject can have a blockage in the lymphatic system, optionally wherein the blockage prevents lymph fluid from draining well, and wherein the fluid buildup leads to swelling. 
     In some methods, the composition is administered by local injection or infusion at or adjacent to a site of interest, for example in one or both arms and/or legs. The site of interest can be a site of lymphatic blockage and/or lymphedema or another site in need of lymphatic system generation or regeneration. The composition can be administered by any suitable means including, but not limited to, intramuscular, intraperitoneal, intravenous, subcutaneous, or subdermal injection or infusion. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG. 1  is a plot showing nanotracking analysis of the size distribution of extracellular vesicles (EV) isolated from conditioned media of adipose-derived stem cells. 
         FIGS. 2A-2C  are bar graphs showing proliferation ( 2 A, WST-8 assay), migration ( 2 B, Boyden chamber assay), and tube length ( 2 C, pixel length per field (×40 magnification, 5 random fields)) of lymphatic endothelial cells (LECs) treated with PBS, HEK 293-EVs, VEGF-C, or adipose-derived stem cells (ADSC)-EVs. For  FIGS. 2A and 2B , values are means (SD) (n=6, triplicate) (*P&lt;0.05). For  FIG. 2C , values are means (SD) (n=6, duplicate) (*P&lt;0.05). 
         FIGS. 3A-3H  are bar graphs showing mRNA expression levels of LYVE-1 ( 3 A,  3 E), VEGFR-3 ( 3 B,  3 F), Prox1 ( 3 C,  3 G) and podoplanin ( 3 D,  3 H) in LECs 12 hrs. ( 3 A- 3 D) or 24 hrs ( 3 E- 3 H) after treatment with PBS, VEGF-C, or ADSC-EVs. The samples were analyzed by qRT-PCR to evaluate the expression of genes. The data were normalized based on GAPDH expression and shown as changes relative to PBS group. Values are means (SD) (n=3, triplicate) (*P&lt;0.05 vs PBS). GAPDH indicates glyceraldehyde-3-phosphate dehydrogenase. 
         FIG. 4A  is a schematic illustrating a splinted lymphedema model. After x-ray irradiation in the bilateral inguinal region at 10 Gy in a single dose twice prior to the surgery, mice were subjected to circumferential incision in the inguinal region. After the resection of inguinal lymph nodes, a 3-mm-wide silicone splint was placed in the inguinal wound and then fixed to the skin and underlying muscle to prevent wound contraction and desiccation. As a therapeutic intervention, injection of PBS or ADSC-EVs (˜40 μg) or HEK293-EVs (˜40 μg) as a thin layer to the whole leg area was performed in postoperative day 7 and 14.  FIG. 4B  is a line graph showing the ratio of hindlimb circumference change over 4 weeks after injections of PBS or ADSC-EVs.  FIG. 4C  is a bar graph showing quantitation of computed tomographic images of indocyanine green (ICG) lymphography used to assess lymphatic function of the hindlimb, after indocyanine green injection of both feet. 
         FIGS. 5A-5C  are bar graphs showing histological analysis of LYVE-1(+) area (%) ( 5 A), CD31(+) area (%) ( 5 B), CD31(+)/LYVE-1(+) overlap area (%) ( 5 C) per 0.25 mm2 area (four random areas per limb) following treatment with PBS, HEK 293-EVs, or ADSC-EVs. All data represent mean±s.d. with P&lt;0.05 considered as significant. 
         FIGS. 6A-6B  are bar graphs showing collagen 1(+) area (%) ( 6 A) and pSMAD3(+) area (%) ( 6 B) per 0.25 mm2 area (four random areas per limb) after treatments of PBS, HEK293-EVs, or ADSC-EVs. All data represent mean±s.d. with P&lt;0.05 considered as significant. 
         FIG. 7  is a gene interaction network map illustrating the results of an Ingenuity Pathway Analysis (IPA) evaluating the implications of miRNA expression found to be altered by ADSC-EVs treatment. This map provides the top ranked network found associated with the role of ADSC-EVs in lymphatic endothelial cells. 
     
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     I. Definitions 
     As used herein, the term “carrier” or “excipient” refers to an organic or inorganic ingredient, natural or synthetic inactive ingredient in a formulation, with which one or more active ingredients are combined. 
     As used herein, the term “pharmaceutically acceptable” means a non-toxic material that does not interfere with the effectiveness of the biological activity of the active ingredients. 
     As used herein, the term “pharmaceutically acceptable carrier” encompasses any of the standard pharmaceutical carriers, such as a phosphate buffered saline solution, water and emulsions such as an oil/water or water/oil emulsion, and various types of wetting agents. As used herein, the terms “effective amount” or “therapeutically effective amount” means a dosage sufficient to alleviate one or more symptoms of a disorder, disease, or condition being treated, or to otherwise provide a desired pharmacologic and/or physiologic effect. The precise dosage will vary according to a variety of factors such as subject-dependent variables (e.g., age, immune system health, etc.), the disease or disorder being treated, as well as the route of administration and the pharmacokinetics of the agent being administered. 
     As used herein, the term “prevention” or “preventing” means to administer a composition to a subject or a system at risk for or having a predisposition for one or more symptom caused by a disease or disorder to cause cessation of a particular symptom of the disease or disorder, a reduction or prevention of one or more symptoms of the disease or disorder, a reduction in the severity of the disease or disorder, the complete ablation of the disease or disorder, stabilization or delay of the development or progression of the disease or disorder. 
     As used herein, the terms “subject,” “individual,” and “patient” refer to any individual who is the target of treatment using the disclosed compositions. The subject can be a vertebrate, for example, a mammal. Thus, the subject can be a human. The subjects can be symptomatic or asymptomatic. The term does not denote a particular age or sex. Thus, adult and newborn subjects, whether male or female, are intended to be covered. A subject can include a control subject or a test subject. As used herein, “substantially changed” means a change of at least e.g. 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, 100%, or more relative to a control. 
     As used herein, the term “purified,” “isolated,” and like terms relate to the isolation of a molecule or compound in a form that is substantially free (at least 60% free, preferably 75% free, and most preferably 90% free) from other components normally associated with the molecule or compound in a native environment. 
     As used herein, the term “antibody” refers to natural or synthetic antibodies that bind a target antigen. The term includes polyclonal and monoclonal antibodies. In addition to intact immunoglobulin molecules, also included in the term “antibodies” are fragments or polymers of those immunoglobulin molecules, and human or humanized versions of immunoglobulin molecules that bind the target antigen. 
     As used herein, “treatment” refers to the medical management of a patient with the intent to cure, ameliorate, stabilize, or prevent a disease, pathological condition, or disorder. This term includes active treatment, that is, treatment directed specifically toward the improvement of a disease, pathological condition, or disorder, and also includes causal treatment, that is, treatment directed toward removal of the cause of the associated disease, pathological condition, or disorder. In addition, this term includes palliative treatment, that is, treatment designed for the relief of symptoms rather than the curing of the disease, pathological condition, or disorder; preventative treatment, that is, treatment directed to minimizing or partially or completely inhibiting the development of the associated disease, pathological condition, or disorder; and supportive treatment, that is, treatment employed to supplement another specific therapy directed toward the improvement of the associated disease, pathological condition, or disorder. 
     As used herein, the terms “inhibit” or “reduce” means to decrease, hinder or restrain a particular characteristic such as an activity, response, condition, disease, or other biological parameter. It is understood that this is typically in relation to some standard or expected value, i.e., it is relative, but that it is not always necessary for the standard or relative value to be referred to. “Inhibits” or “reduce” can also mean to hinder or restrain the synthesis, expression or function of a protein relative to a standard or control. This can include, but is not limited to, the complete ablation of the activity, response, condition, or disease. Inhibition may also include, for example, a 10% reduction in the activity, response, condition, disease, or other biological parameter as compared to the native or control level. Thus, the reduction can be about 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, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64,65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100%, or any amount of reduction in between as compared to native or control levels. 
     Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. Use of the term “about” is intended to describe values either above or below the stated value in a range of approx. +/−10%; in other forms the values may range in value either above or below the stated value in a range of approx. +/−5%; in other forms the values may range in value either above or below the stated value in a range of approx. +/−2%; in other forms the values may range in value either above or below the stated value in a range of approx. +/−1%. The preceding ranges are intended to be made clear by context, and no further limitation is implied. 
     II. Compositions 
     In general, cell-based therapies have limitations such as uncontrolled differentiation, side effects, tumor formation, and incompatibility of allogenic use. On the contrary, regenerative therapy using extracellular vesicles (EVs) from MSCs have the possibility to overcome such disadvantages cell-based therapies have. 
     The conditioned medium which contains cultured MSCs secretion have also been reported to have lymphangiogenic effects and therapeutic potential (Takeda et al.,  Ann Plast Surg.,  June;74(6):728-36 (2015)). MSCs can secrete cytokines, chemokines, growth factors, and EVs (Katsuda et al.,  Proteomics,  May;13(10-11) (2013)). These EVs are produced by a variety of cell types and may pay a role as intercellular transmitters of mRNA, microRNA, and proteins (Valadi et al.,  Nature Cell Biology  volume 9, pages 654-659 (2007)). There is some evidence indicating that some of the regenerative properties previously credited to MSCs may be related to the secreted EVs (Lai et al.,  Stem Cell Res.,  May;4(3):214-22 (2010), Bruno et al.,  J Am Soc Nephrol.,  May;20(5):1053-67 (2009)). However, the experiments discussed in the Examples below investigate the therapeutic ability of EVs secreted from ADSCs using in vitro and in vivo experimental systems and demonstrate that ADSC-EVs can modulate the phenotype of lymphatic endothelial cells (LECs), which may contribute to several lymphatic pathophysiological processes. Results further demonstrate the therapeutic potential of ADSC-EVs in relief of lymphedema using a mouse hindlimb lymphedema model. Cell-free compositions including EVs and methods of use thereof are provided. The EVs can be part of a heterogeneous mixture of factors such as conditioned media, or a fraction isolated therefrom. In other embodiments, EVs, or one or more subtypes thereof, are isolated or otherwise collected from conditioned media. The EVs, or one or more subtypes thereof, can be suspended in a pharmaceutically acceptable composition, such as a carrier or matrix or depot, prior to administration to the subject. 
     A. Extracellular Vesicles 
     The disclosed compositions typically are or include extracellular vesicles derived from mesenchymal cells, or an isolated or fractionated subtype or subtypes thereof. Extracellular vesicles are lipid bilayer-delimited particles that are naturally released from a cell and, unlike a cell, cannot replicate. EVs range in diameter from near the size of the smallest physically possible unilamellar liposome (around 20-30 nanometers) to as large as 10 microns or more, although the vast majority of EVs are smaller than 200 nm. EVs secreted from MSCs play a role in MSC-mediated paracrine effects via transfer of miRNAs (Spees et al.,  Stem Cell Res Ther.,  7:125 (2016)), and may have effects on wound healing (Zhang et al.,  J Transl Med., February  1;13:49 (2015)), skin rejuvenation (Kim et al.,  Biochem Biophys Res Commun., November  18;493(2):1102-1108 (2017)), angiogenesis (Gong et al.,  Oncotarget, July  11;8(28):45200-45212 (2017)), adjusting immunologic function (Zhao et al.,  Diabetes,  February;67(2):235-247 (2018)), regeneration of damaged tissue (Lai et al.,  Stem Cell Res.,  May;4(3):214-22 (2010)) as well as relief neurological disorders (Katsuda et al.,  Proteomics,  May;13(10-11) (2013)). Diverse EV subtypes have been proposed including ectosomes, microvesicles (MV), microparticles, exosomes, oncosomes, apoptotic bodies (AB), tunneling nanotubes (TNT), and more (Yáñez-Mó, et al.,  J Extracell Vesicles.  4: 27066 (2015) doi:10.3402/jev.v4.27066. PMC 4433489). These EV subtypes have been defined by various, often overlapping, definitions, based mostly on biogenesis (cell pathway, cell or tissue identity, condition of origin) (Théry, et al.,  J Extracell Vesicles.  7 (1): 1535750 (2018). doi:10.1080/20013078.2018.1535750). However, EV subtypes may also be defined by size, constituent molecules, function, or method of separation. According to minimal information from studies of extracellular vesicles 2018 (MISEV2018)12, EV is a collective term covering various subtypes of cell-released, membranous structures, called exosomes, microvesicles, microparticles, ectosomes, oncosomes, apoptotic bodies, and many other names (Clotilde, et al.,  J Extracell Vesicles,  7(1):1535750 (2018)). As discussed in Théry, et al., subtypes of EVs may be defined by:
         a) physical characteristics of EVs, such as size (“small EVs” (sEVs) and “medium/large EVs” (m/lEVs), with ranges defined, for instance, respectively, &lt;100 nm or &lt;200 nm [small], or &gt;200 nm [large and/or medium]) or density (low, middle, high, with each range defined);   b) biochemical composition (CD63+/CD81+−EVs, Annexin A5-stained EVs, etc.); or   c) descriptions of conditions or cell of origin (podocyte EVs, hypoxic EVs, large oncosomes, apoptotic bodies).       

     Thus, in some embodiments, the composition is or includes one or more EV subtypes defined according (a), (b), or (c) as discussed above. 
     In some embodiments, the vesicles are or include exosomes. Exosomes possess the surface proteins that promote endocytosis and they have the potential to deliver macromolecules. Also, if the exosomes are obtained from the same individual as they are delivered to, the exosomes will be immunotolerant. Due to the technical limitations, previous studies are not sufficient to conclude that exosomes have specific functions compared with other EVs (Clotilde, et al.,  J Extracell Vesicles,  7(1):1535750 (2018)). 
     Exosomes are vesicles with the size of 30-150 nm, often 40-100 nm, and are observed in most cell types. Exosomes are often similar to MVs with an important difference: instead of originating directly from the plasma membrane, they are generated by inward budding into multivesicular bodies (MVBs). The formation of exosomes includes three different stages: (1) the formation of endocytic vesicles from plasma membrane, (2) the inward budding of the endosomal vesicle membrane resulting in MVBs that consist of intraluminal vesicles (ILVs), and (3) the fusion of these MVBs with the plasma membrane, which releases the vesicular contents, known as exosomes. 
     Exosomes have a lipid bilayer with an average thickness of ˜5 nm (see e.g., Li,  Theranostics,  7(3):789-804 (2017) doi: 10.7150/thno.18133). 
     The lipid components of exosomes include ceramide (sometimes used to differentiate exosomes from lysosomes), cholesterol, sphingolipids, and phosphoglycerides with long and saturated fatty-acyl chains. The outer surface of exosomes is typically rich in saccharide chains, such as mannose, polylactosamine, alpha-2,6 sialic acid, and N-linked glycans. 
     Many exosomes contain proteins such as platelet derived growth factor receptor, lactadherin, transmembrane proteins and lysosome associated membrane protein-2B, membrane transport and fusion proteins like annexins, flotillins, GTPases, heat shock proteins, tetraspanins, proteins involved in multivesicular body biogenesis, as well as lipid-related proteins and phospholipases. These characteristic proteins therefore serve as good biomarkers for the isolation and quantification of exosomes. Another key cargo that exosomes carry is nucleic acids including deoxynucleic acids (DNA), coding and non-coding ribonucleic acid (RNA) like messenger RNA (mRNA) and microRNA (miRNA). 
     In some embodiments, the vesicles include or are one or more alternative extracellular vesicles, such as ABs, MVs, TNTs, or others discussed herein or elsewhere. 
     ABs are heterogenous in size and originate from the plasma membrane. They can be released from all cell types and are about 1-5 μm in size. 
     MVs with the size of 20 nm-1 μm are formed due to blebbing with incorporation of cytosolic proteins. In contrast to ABs, the shape of MVs is homogenous. They originate from the plasma membrane and are observed in most cell types. 
     TNTs are thin (e.g., 50-700 nm) and up to 100 μm long actin containing tubes formed from the plasma membrane. 
     The Examples below show that EVs isolated from ADSC were physically homogeneous with a peak of 100 nm ( FIG. 1 ). Thus, in some embodiments, the EVs are between about 20 nm and about 500 nm. In some embodiments, the EVs are between about 20 nm and about 250 nm or 200 nm or 150 nm or 100 nm. 
     The Examples below show that EVs isolated from ADSC include CD9 and CD63. Thus, in some embodiments, the EVs include CD9, CD63, or both. 
     The Examples below show that EVs isolated from ADSC include a number of miRNAs. In some embodiments, the EVs include one or more of the miRNA of Table 1. In some embodiments, the EVs include on or more of miR-199a-3p, miR-145-5p, miR-143-3p, miR-377-3p, miR-100-3p, miR-29a-3p, miR-495-3p, miR-29c-3p, miR-658, miR-493-3p, miR-184, and miR-27a-3p. 
     B. Methods of Making Extracellular Vesicles 
     1. Sources of Cells for Making Extracellular Vesicles 
     As used herein, EVs, including AB, MV, exosomes, and TNT typically refer to lipid vesicles formed by cells or tissue. They can be isolated from tissue, cells, and fluid directly from a subject, including cultured and uncultured tissue, cells, or fluids, and fluid derived or conditioned by cultured cells (e.g., conditioned media). For example, exosomes are present in physiological fluids such as plasma, lymph liquid, malignant pleural effusion, amniotic liquid, breast milk, semen, saliva and urine, and are secreted into the media of cultured cells. 
     The EVs of the disclosed compositions are typically formed from mesenchymal cells, preferably mesenchymal stem cells. Mesenchymal stem cells (MSCs) are multipotent adult stem cells that are present in multiple tissues, including umbilical cord, bone marrow and fat tissue. Mesenchymal stem cells can self-renew by dividing and can differentiate into multiple tissues including bone, cartilage, muscle and fat cells, and connective tissue. Thus, mesenchymal cells include, for example, adipocytes, chondrocytes, osteoblasts, myocytes and tendon, and MSCs are multipotent stem cells that can differentiate in one or more of these cell types. 
     In some embodiments, the EVs are formed by MSCs. MSCs can be derived from bone barrow or other non-marrow tissues, such as placenta, umbilical cord blood, adipose tissue, adult muscle, corneal stroma or the dental pulp of deciduous (baby) teeth. 
     In some embodiments, the MSCs are derived from adipose tissue. Adipose tissue-derived MSCs (AdMSCs or ADSC), in addition to being easier and safer to isolate than bone marrow-derived MSCs, can be obtained in larger quantities. 
     Methods of isolating extracellular vesicles from tissue, cells, and fluid directly from a subject, including cultured and uncultured tissue, cells, or fluids, and fluid derived or conditioned by cultured cells (e.g., conditioned media) are known in the art. 
     See, for example, Li,  Themaostics,  7(3):789-804 (2017) doi: 10.7150/thno.18133, Ha, et al.,  Acta Pharmaceutica Sinica B,  6(4):287-296 (2016) doi: 10.1016/j.apsb.2016.02.001, Skotland, et al.,  Progress in Lipid Research,  66:30-41 (2017) doi: 10.1016/j.plipres.2017.03.001, Phinney and Pittenger,  Stem Cells,  35:851-858 (2017) doi: 10.1002/stem.2575, each of which is specifically incorporated by reference, and describes isolating extracellular vesicles, particularly exosomes. 
     The EVs can be collected from primary cells or tissue or fluid. In some embodiments, the vesicles are isolated from cells, tissue, or fluid of the subject to be treated. An advantage of utilizing EVs that are isolated from natural sources includes avoidance of immunogenicity that can be associated with artificially produced lipid vesicles. 
     The EVs can also be collected from cell lines or tissue. Exemplary cells lines are commercially available and include those various sources including human bone-marrow, human umbilical cord, human embryonic tissue, and human adipose including those derived from lipoaspirate or dedifferentiated from mature adipocytes. 
     2. Methods of Collecting Extracellular Vesicles 
     Extracellular vesicles, including exosomes, can be isolated using differential centrifugation, flotation density gradient centrifugation, filtration, high performance liquid chromatography, and immunoaffinity-capture. 
     For example, one of the most common isolation techniques for isolating exosomes from cell culture is differential centrifugation, whereby large particles and cell debris in the culture medium are separated using centrifugal force between 200-100,000×g and the exosomes are separated from supernatant by the sedimenting exosomes at about 100,000×g. Purity can be improved, however, by centrifuging the samples using flotation density gradient centrifugation with sucrose or Optiprep. Tangential flow filtration combined with deuterium/sucrose-based density gradient ultracentrifugation was employed to isolate therapeutic exosomes for clinical trials. 
     In the experiments provided below, EVs were isolated from ADSCs. After incubation for two days, the medium was collected and centrifuged at 2,000 g for 15 min at room temperature. To thoroughly remove cellular debris, the supernatant was filtered with a 0.22-mm filter unit. Then, the cultured media (CM) was ultracentrifuged at 110,000 g (35,000 rpm) for 70 min at 4° C. 
     Ultrafiltration and high-performance liquid chromatography (HPLC) are additional methods of isolating EVs based on their size differences. EVs prepared by HPLC are highly purified. 
     Hydrostatic filtration dialysis has been used for isolating extracellular vesicles from urine. 
     Other common techniques for EV collection involve positive and/or negative selection using affinity-based methodology. Antibodies can be immobilized in different media conditions and combined with magnetic beads, chromatographic matrix, plates, and microfluidic devices for separation. For example, antibodies against exosome-associated antigens—such as cluster of differentiation (CD) molecules CD63, CD81, CD82, CD9, epithelial cell adhesion molecule (EpCAM), and Ras-related protein (Rab5)—can be used for affinity-based separation of exosomes. Non-exosomes vesicles that carry these or different antigens can also be isolated in a similar way. 
     Microfluidics-based devices have also been used to rapidly and efficiently isolate EVs such as exosomes, tapping on both the physical and biochemical properties of exosomes at microscales. In addition to size, density, and immunoaffinity, sorting mechanisms such as acoustic, electrophoretic and electromagnetic manipulations can be implemented. 
     Methods of characterizing EVs including exosomes are also known in the art. Exosomes can be characterized based on their size, protein content, and lipid content. Exosomes are sphere-shaped structures with sizes between 40-100 nm and are much smaller compared to other systems, such as a microvesicle, which has a size range from 100-500 nm. Several methods can be used to characterize EVs, including flow cytometry, nanoparticle tracking analysis, dynamic light scattering, western blot, mass spectrometry, and microscopy techniques. EVs can also be characterized and marked based on their protein compositions. For example, integrins and tetraspanins are two of the most abundant proteins found in exosomes. Other protein markers include TSG101, ALG-2 interacting protein X (ALIX), flotillin 1, and cell adhesion molecules. Similar to proteins, lipids are major components of EVs and can be utilized to characterize them. 
     C. Pharmaceutical Compositions 
     Pharmaceutical compositions including EVs are also provided. Pharmaceutical compositions can be administered parenterally (intramuscular (IM), intraperitoneal (IP), intravenous (IV), subcutaneous injection (SubQ), subdermal), transdermally (either passively or using iontophoresis or electroporation), or by any other suitable means, and can be formulated in dosage forms appropriate for each route of administration. 
     In some embodiments, the compositions are administered systemically, for example, by intravenous or intraperitoneal administration, in an amount effective for delivery of the compositions to targeted cells. 
     In preferred embodiments, the compositions are administered locally, for example, by injection directly into, or adjacent to, a site to be treated. For example, in some embodiments such as for the treatment of lymphedema, the compositions are injected or otherwise administered directly to the lymphedemic area or the area adjacent thereto (e.g., in the arms or legs). 
     Typically, local injection causes an increased localized concentration of the compositions which is greater than that which can be achieved by systemic administration. 
     In some embodiments, the compositions are delivered locally to the appropriate cells by using a catheter or syringe. Other means of delivering such compositions locally to cells include using infusion pumps (for example, from Alza Corporation, Palo Alto, Calif.) or incorporating the compositions into polymeric implants (see, for example, P. Johnson and J. G. Lloyd-Jones, eds.,  Drug Delivery Systems: Fundamentals and Techniques  (Chichester, England: Ellis Horwood Ltd., 1988 ISBN-10: 0895735806), which can effect a sustained release of the material to the immediate area of the implant. 
     The EV compositions can be provided to the cell either directly, such as by contacting it with the cell, or indirectly, such as through the action of any biological process. For example, the vesicles can be formulated in a physiologically acceptable carrier, and injected into a tissue or fluid surrounding the cell. 
     Exemplary dosage for in vivo methods are discussed in the experiments below. As further studies are conducted, information will emerge regarding appropriate dosage levels for treatment of various conditions in various patients, and the ordinary skilled worker, considering the therapeutic context, age, and general health of the recipient, will be able to ascertain proper dosing. The selected dosage depends upon the desired therapeutic effect, on the route of administration, and on the duration of the treatment desired. 
     Generally, for local injection or infusion, dosage may be lower. Generally, the total amount of the active agent administered to an individual using the disclosed vesicles can be less than the amount of unassociated active agent that must be administered for the same desired or intended effect and/or may exhibit reduced toxicity. 
     In a preferred embodiment the compositions are administered in an aqueous solution, by parenteral injection such as intramuscular, intraperitoneal, intravenous, subcutaneous, subdermal, etc. 
     The formulation can be in the form of a suspension or emulsion. In general, pharmaceutical compositions are provided including effective amounts of one or more active agents optionally include pharmaceutically acceptable diluents, preservatives, solubilizers, emulsifiers, adjuvants and/or carriers. Such compositions can include diluents, sterile water, buffered saline of various buffer content (e.g., Tris-HCl, acetate, phosphate) at various pHs and ionic strengths; and optionally, additives such as detergents and solubilizing agents (e.g., TWEEN® 20, TWEEN® 80 also referred to as polysorbate 20 or 80), anti-oxidants (e.g., ascorbic acid, sodium metabisulfite), and preservatives (e.g., Thimersol, benzyl alcohol) and bulking substances (e.g., lactose, mannitol). Examples of non-aqueous solvents or vehicles are propylene glycol, polyethylene glycol, vegetable oils, such as olive oil and corn oil, gelatin, and injectable organic esters such as ethyl oleate. The formulations may be lyophilized and redissolved/resuspended immediately before use. The formulation may be sterilized by, for example, filtration through a bacterium retaining filter, by incorporating sterilizing agents into the compositions, by irradiating the compositions, or by heating the compositions. 
     Transdermal formulations may also be prepared. These will typically be ointments, lotions, sprays, or patches, all of which can be prepared using standard technology. Transdermal formulations can include penetration enhancers. Chemical enhancers and physical methods including electroporation and microneedles can work in conjunction with this method. 
     III. Methods of Use 
     Methods of using the disclosed compositions are also provided. The experiments below illustrate that EVs from ADSCs promoted proliferation, migration, and tube formation activities, and upregulated gene expression of lymphatic markers in lymphatic endothelial cells. ADSC-EVs increased both LYVE-1 positive lymphatic endothelial cells and CD-31 positive vascular endothelial cells of injected limbs in vivo, and suppressed immunologic activities. These alterations induced by ADSC-EVs resulted in relief the condition of lymphedema. 
     Thus, in some embodiments, the disclosed compositions are administered to a subject in need thereof in an effective amount. In some embodiments, the amount is effective to increase the proliferation, migration, and/or tube formation of lymphatic endothelial cells, increase expression of one or more lymphatic makers (e.g., hyaluronan receptor-1 (LYVE-1), vascular endothelial growth factor receptor-3 (VEGFR-3), prospero homeobox 1 (Prox1), and/or podoplanin) in lymphatic endothelial cells, increase angiogenesis, increase lymphangiogeneisis, reduce inflammatory response, decrease fibrosis formation, enlarging circumference and/or inducing formation of capillary vessels and/or lymphatic vessels, induce formation of vessels that express both vascular and lymphatic markers, increase drainage routes (e.g., for accumulated fluids), increase HIF1-alpha expression and/or activity (e.g., in lymphatic endothelial cells), reduce Prohibitin (PHB) expression and/or activity (e.g., in lymphatic endothelial cells), or a combination thereof in a subject. 
     The disclosed compositions and methods are particularly useful for treating a subject having lymphedema, and/or symptoms associated therewith. Thus, in some embodiments, EVs or a subtype or subtypes thereof or a composition thereof, are administered to a subject in need thereof in an effective amount to treat lymphedema, or one or more symptoms associated therewith. 
     Lymphedema refers to swelling that generally occurs in one or both arms or legs. Lymphedema is generally caused by blockage the lymphatic system. The blockage prevents lymph fluid from draining well, and the fluid buildup leads to swelling. 
     Lymphedema signs and symptoms, which typically occur in the affected arm(s) and/or leg(s), include: swelling of part or all of the arm(s) and/or leg(s), including fingers or toes, a feeling of heaviness or tightness, restricted range of motion, aching or discomfort, recurring infections, hardening and thickening of the skin (fibrosis). 
     Draining excess lymphatic fluid is one of the main purposes of lymphedema treatment. To promote remaining drainage function, traditional compression care or microsurgical lymphatic system reconstruction has been performed for lymphedema management (Tashiro, et al.,  J Plast Reconstr Aesthet Surg.,  69(3):368-75 (2016)). Recent advancement of surgical approach to lymphedema has been attempted to create lympho-venous shunts or healthy lymph nodes transplantation from other sites. Thus, in some embodiments, the disclosed methods are combined with one or more of these or other conventional methods of treating lymphedema. 
     In the studies below, the effects of ADSC-EVs treatment were investigated both in vitro and in vivo situations, and results indicate that inducing both angiogenesis and lymphangiogenesis may lead to the establishment of new connection between capillary vessels and lymphatic vessels. The existence of the route between capillary system and lymphatic system may work lifelong as drainage route of excess lymphatic fluid in affected tissues as a treatment option. 
     Lymphedema can be caused by the removal of, or damage to, lymph nodes as a part of cancer treatment. Thus, in some embodiments, the subject also has cancer. In some embodiments, the subject does not have cancer. 
     In some embodiments, the EVs are administered as part of a heterogeneous mixture of factors (e.g., conditioned media, or a fraction isolated therefrom). In some embodiments, EVs or more of more subtypes thereof are isolated or otherwise collected from conditioned media. The EVs or one or more subtypes thereof can be suspended in pharmaceutically acceptable composition, such as a carrier or matrix or depot, prior to administration to the subject. 
     EVs may possess the versatility and capacity to interact with multiple cell types immediately and remote areas to regulate cellular responses (Zhang et al.,  Cell Prolif.,  49:3-13 (2016)). Thus, although regional or local administration to the site of interest (e.g., the site of lymphedema) or a site adjacent thereto is preferred, systemic administration is also contemplated. Furthermore, although lymphatic endothelial cells are a preferred target, the EVs may also affect other cells in the region of administration that effect the treatment outcome. 
     The frequency of administration of a method of treatment can be, for example, one, two, three, four or more times daily, weekly, every two weeks, or monthly. In some embodiments, the composition is administered to a subject once every 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, or 31 days. In some embodiments, the frequency of administration is once, twice or three times weekly, or is once, twice or three times every two weeks, or is once, twice or three times every four weeks. In some embodiments, the composition is administered to a subject 1-3 times, preferably 2 times, a week. 
     In some embodiments, the effect of the disclosed compositions and methods on a subject is compared to a control. For example, the effect of the composition on a particular symptom, pharmacologic, or physiologic indicator (including those mentioned above and elsewhere herein) can be compared to an untreated subject, or the condition of the subject prior to treatment. In some embodiments, the symptom, pharmacologic, or physiologic indicator is measured in a subject prior to treatment, and again one or more times after treatment is initiated. In some embodiments, the control is a reference level, or average determined based on measuring the symptom, pharmacologic, or physiologic indicator in one or more subjects that do not have the disease or condition to be treated (e.g., healthy subjects). 
     In some embodiments, the effect of the treatment is compared to a conventional treatment that is known in the art, such as one of those discussed herein. 
     IV. Kits 
     Dosage units including the disclosed compositions, for example, in a pharmaceutically acceptable carrier for shipping and storage and/or administration are also disclosed. Components of the kit may be packaged individually and can be sterile. In some embodiments, a pharmaceutically acceptable carrier containing an effective amount of the composition is shipped and stored in a sterile vial. The sterile vial may contain enough composition for one or more doses. The composition may be shipped and stored in a volume suitable for administration, or may be provided in a concentration that is diluted prior to administration. In another embodiment, a pharmaceutically acceptable carrier containing drug can be shipped and stored in a syringe. 
     Kits containing syringes of various capacities or vessels with deformable sides (e.g., plastic vessels or plastic-sided vessels) that can be squeezed to force a liquid composition out of an orifice are provided. The size and design of the syringe will depend on the route of administration. Any of the kits can include instructions for use. 
     The disclosed compositions and methods can be further understood through the following numbered paragraphs. 
     1. A method of promoting generation or regeneration of the lymphatic system in a subject comprising administering the subject a composition comprising an effective amount of extracellular vesicles formed by mesenchymal stem cells (MSCs) to increase generation of the lymphatic system. 2. A method of treating a subject for lymphedema comprising administering the subject an effective amount of a composition comprising extracellular vesicles formed by mesenchymal stem cells to increase generation of the lymphatic system. 
     3. The method of paragraphs 1 or 2, wherein the composition comprises a pharmaceutically acceptable carrier. 
     4. The method of any one of paragraphs 1-3, wherein the composition is cell-free. 
     5. The method of any one of paragraphs 1-4, wherein the extracellular vesicles are formed by a method comprising culturing MSCs to produce media conditioned with the extracellular vesicles. 
     6. The method of paragraph 5, wherein the method further comprises separating extracellular vesicles from the media conditioned by the MSCs. 
     7. The method of paragraph 6, wherein the composition does not comprise the media conditioned by the MSCs. 
     8. The method of any one of paragraphs 1-7, wherein the MSCs are primary cells or a cell line. 
     9. The method of any one of paragraphs 1-8, wherein the MSCs are from bone barrow, placenta, umbilical cord blood, adipose tissue, adult muscle, conical stroma, or the dental pulp of deciduous teeth. 
     10. The method of any one of paragraphs 1-9, wherein the MSCs are adipose-derived stem cells. 
     11. The method of any one of paragraphs 1-10, wherein the extracellular vesicles comprise or consist of ectosomes, microvesicles (MV), microparticles, exosomes, oncosomes, apoptotic bodies (AB), tunneling nanotubes (TNT), or a combination thereof. 
     12. The method of paragraph 11, wherein the extracellular vesicles comprise or consist of exosomes, microvesicles or a combination thereof. 
     13. The method of any one of paragraphs 1-12, wherein the extracellular vesicles comprise or consist of a vesicles having a size of between about 20 nm and about 500 nm, or between about 20 nm and about 250 nm, or between about 20 nm and about 200 nm, or between about 20 nm and about 150 nm, or between about 20 nm and about 100 nm. 
     14. The method of any one of paragraphs 1-13, wherein the extracellular vesicles comprise CD9, CD63, or a combination thereof 15. The method of any one of paragraphs 1-14, wherein the extracellular vesicles comprise one or more of miR-199a-3p, miR-145-5p, miR-143-3p, miR-377-3p, miR-100-3p, miR-29a-3p, miR-495-3p, miR-29c-3p, miR-658, miR-493-3p, miR-184, and miR-27a-3p. 
     16. The method of any one of paragraphs 1-15 comprising increasing the proliferation, migration, and/or tube formation of lymphatic endothelial cells, increasing expression of one or more lymphatic markers (e.g., hyaluronan receptor-1(LYVE-1), vascular endothelial growth factor receptor-3 (VEGFR-3), prospero homeobox 1 (Prox1), and/or podoplanin) in lymphatic endothelial cells, increasing angiogenesis, increasing lymphangiogeneisis, reducing inflammatory response, decreasing fibrosis formation, enlarging circumference and/or inducing formation of capillary vessels and/or lymphatic vessels, inducing formation of vessels that express both vascular and lymphatic markers, increasing drainage routes (e.g., for accumulated fluids), increasing HIF1-alpha expression and/or activity, reducing Prohibitin (PHB) expression and/or activity, or a combination thereof in the subject. 
     17. The method of any one of paragraphs 1-16, wherein the subject has a blockage in the lymphatic system, optionally wherein the blockage prevents lymph fluid from draining well, and wherein the fluid buildup leads to swelling. 
     18. The method of any one of paragraphs 1-17, wherein the subject has one or more symptoms selected from swelling of part or all of the arm(s) and/or leg(s), a feeling of heaviness or tightness, restricted range of motion, aching or discomfort, recurring infections, and fibrosis in one or both arms and/or legs. 
     19. The method of any one of paragraphs 1-18, wherein the subject has been diagnosed with lymphedema. 
     20. The method of any one of paragraphs 1-19, wherein the composition is administered by local injection or infusion at or adjacent to a site of interest. 
     21. The method of paragraph 20, wherein the site of interest is in one or both arms and/or legs. 
     22. The method of paragraphs 20 or 21, wherein the site of interest is a site of lymphatic blockage and/or lymphedema. 
     23. The method of any one of paragraphs 1-22, wherein the composition is administered by intramuscular, intraperitoneal, intravenous, subcutaneous, or subdermal injection. 
     24. The composition of any one of paragraphs 1-23. 
     25. A composition comprising an effective amount of extracellular vesicles formed by mesenchymal stem cells (MSCs) suitable for use in the method of any one of paragraphs 1-23. 
     26. Use of the composition of paragraph 24 or 25 for promoting generation or regeneration of the lymphatic system. 
     27. Use of the composition of paragraph 24 or 25 for the manufacture of a medicament for promoting generation or regeneration of the lymphatic system. 
     28. Use of the composition of paragraph 24 or 25 for treating lymphedema. 
     29. Use of the composition of paragraph 24 or 25 for the manufacture of a medicament for treating lymphedema. 
     30. A composition, use, or method according to any of the disclosure herein including, but not limited to, the description, the experimental examples, and/or the figures and their descriptions. 
     EXAMPLES 
     Example 1 
     Characteristics of ADSC-Derived EVs Materials and Methods 
     Cell Culture and Preparation of EVs 
     Human adipose derived stem cells were purchased from Lonza (Basel, Switzerland) and cultured in Dulbecco&#39;s Modified Eagle Medium (DMEM; Nissui Pharmaceutical Co, Tokyo, Japan) supplemented with 10% fetal bovine serum. Primary cells were cultured for 7 days (passage 0), replacing the medium three times weekly. Cell passage was done each week in 0.25% trypsin/2 mM EDTA (37° C., 5 min). All ADSCs were used within the sixth passage. 
     At approximate 80% confluence, ADSCs were washed with PBS thrice and the culture media were replaced with DMEM containing 0.1% fetal bovine serum. After incubation for two days, the medium was collected and centrifuged at 2,000 g for 15 mM at room temperature. To thoroughly remove cellular debris, the supernatant was filtered with a 0.22-mm filter unit (Millipore). Then, the conditioned media (CM) was ultracentrifuged at 110,000 g (35,000 rpm) for 70 mM at 4° C. using an SW41 rotor (Beckman) (Clotilde, et al.,  J Extracell Vesicles,  7(1):1535750 (2018)). The pellets were washed with 11 ml PBS, and after ultracentrifugation, they were resuspended in PBS. The protein concentration of the putative EV fraction was determined using a Quant-iT Protein Assay with a Qubit 2.0 Fluorometer (Invitrogen). To determine the size distribution of the EVs, nanoparticle tracking analysis was carried out using the Nanosight system (NanoSight) on samples diluted 500- to 1,000-fold with PBS for analysis. The system focuses a laser beam through a suspension of the particles of interest. These are visualized by light scattering using a conventional optical microscope perpendicularly aligned to the beam axis, which collects light scattered from every particle in the field of view. A 60 s video recorded all events for further analysis by the nanoparticle tracking analysis software. The Brownian motion of each particle was tracked between frames to calculate its size using the Stokes-Einstein equation. Using same procedures and culture media, EVs derived from Human Embryonic Kidney 293 (HEK293) cells (KAC co ltd, Japan) were obtained. 
     PKH67-Labelled EVs Transfer 
     Purified EVs derived from ADSCs were labelled with a PKH67 green fluorescence labelling kit (Sigma-Aldrich, Mo., USA). EVs were incubated with 2 mM of PKH67 for 5 min and washed five times using a 100 kDa filter (Microcon YM-100, Millipore) to remove excess dye. PKH67-labelled EVs were used to assess EV uptake in vitro. 
     Results 
     Recently, it has been shown that EVs secreted by ADSCs contribute to their paracrine effects. To identify the angiogenic and lymphangiogenic capacity of ADSC-EVs, putative EV fractions were isolated from conditioned media of ADSCs. ADSC-EVs exhibited the characteristic round morphology with bilayer structure under a transmission electron microscope. Nano tracking analysis showed that the size distribution of the isolated EVs was physically homogeneous with a peak diameter of 82 nm ( FIG. 1 ). 
     Immunoblot analyses showed that tetraspanin CD9 and CD63, reliable exosomal markers, were present in the EV fraction. Fluorescence microscopy analysis demonstrated that the PKH67-labelled EVs had been taken up and was transferred to perinuclear compartments, presumably representative of late endocytic compartments. 
     Example 2 
     ADSC-Derived EVs have Lymphangiogenic Effects 
     Materials and Methods 
     LEC Culture 
     Human dermal lymphatic microvascular endothelial cells (HMVEC-dLy Ad) were purchased from Lonza (Basel, Switzerland) and cultured in endothelial growth medium-2-MV(EGM-2-MV; Lonza) that consisted of endothelial basal medium-2 (EBM-2; Lonza) supplemented with 5% fetal bovine serum (FBS), human basic fibroblast growth factor (bFGF), human 
     VEGF, human insulin like growth factor-1 (IGF-1), human epidermal growth factor, hydrocortisone, ascorbic acid, and gentamicin and amphotericin (SingleQuots; Lonza), according to the manufacturer&#39;s instructions. Lymphatic endothelial cells between passages 3 and 6 were used for all experiments in this study. 
     Proliferation Assay 
     LEC proliferation assays were performed as previously described (Takeda, et al., Ann Plast Surg., 74(6):728-36 (2015)) LECs were treated with 100 μL of EBM-2 containing PBS, 10 ng/ml recombinant human VEGF-C (rVEGF-C) (R&amp;D Systems, Minneapolis, Minn.), HEK293-EVs, or ADSC-EVs (10 μg/ml) for 48 hours at 37° C. in 5% CO 2 . Cell proliferation activity was measured using Cell Counting Kit-8 (Dojindo, Kumamoto, Japan), in which 10 μL of the WST-8 assay solution was added to each well and incubated for 4 hours. The absorbance was measured using a microplate reader (Bio-Rad, Hercules, Calif.) at a wavelength of 450 nm. 
     Migration Assay 
     LEC migration assays were performed using Transwell chambers with inserts with 8-μm pores (Corning Costar) as previously described (Takeda, et al.,  Ann Plast Surg.,  74(6):728-36 (2015)). The lower chambers were filled with 700 μL of EBM-2 containing PBS, 10 ng/ml rVEGF-C, HEK293-EVs, or ADSC-EVs (10 μg/ml). After incubation for 16 hours at 37° C. in 5% CO2, the cells on the lower surface of the filter were stained using Diff-Quik (Sysmex, Hyogo, Japan).LECs were photographed through the pores at 100x magnification in 10 random fields, and the migrated cells were counted. 
     Tube Formation Assay 
     Matrigel tube formation assays were carried out as previously described (Takeda, et al.,  Ann Plast Surg.,  74(6):728-36 (2015)). LECs were seeded onto the coated wells and cultured in 500 μL of EBM-2 containing PBS, 10 ng/ml rVEGF-C, HEK293-EVs, or ADSC-EVs (10 μg/ml). After incubation for 8 hours at 37° C. in 5% CO 2 , tube formation images were captured at 40× magnification in 5 random fields. For quantification, the tube length was measured using NIH ImageJ software. 
     Statistical Analysis 
     All statistical analyses were performed using GraphPad Prism 8 (GraphPad Software, Inc., La Jolla, Calif.). All data were expressed as mean±SEM, with the results in the three groups compared by one-way analysis of variance for more than three groups and t test for two groups, with a value of p&lt;0.05 considered significant. 
     Results 
     To investigate the ability of ADSC-EVs to enhance lymphangiogenesis in vitro, the proliferation, migration, and tube formation was investigated in LECs, after addition of ADSC-EVs. Proliferation assay showed that ADSC-EVs markedly increased proliferation (absorbance: 1.51+−0.21, P&lt;0.05) by up to one and one-half times compared with PBS treated group (absorbance: 1.095+−0.11). Similar tendency was observed in VEGF-C treated group (absorbance: 1.62+−0.19, P&lt;0.05), although HEK293-EVs treated group showed no significant changes (absorbance: 1.15+−0.20, P&gt;0.05) ( FIG. 2A ). Migration assay showed that ADSC-EVs significantly promoted LECs migration (120.1+−12.2, cells/field, P&lt;0.05) compared with PBS treated group (90.2 +−11.2, cells/field), as well as VEGF-C treated group (125.3+−11.7, cells/field, P&lt;0.05). HEK293-EVs did not show a significant difference (102.1+−13.3, cells/field, P&gt;0.05) ( FIG. 2B ). The role of ADSC-EVs was further assessed in the regulation of LECs tube formation. Morphological analysis showed ADSC-EVs promote tube formation (2620.5+−321.3, pixel/field, P&lt;0.05) of LECs compared with PBS treated group (1317.8+−500.8 pixel/field) ( FIG. 2C ). VEGF-C treated group showed a significant difference (2593.4+−156.2, pixel/field, P&lt;0.05), however, HEK293-EVs showed no significant difference (1702.1+−280.3, pixel/field, P&gt;0.05) ( FIG. 2C ) These results indicate that ADSC-EVs have lymphangiogenic molecules such as cytokines or miRNAs that stimulated proliferation, migration, and tube formation of LECs. 
     Example 3 
     qRT-PCR Analysis Shows Increased Expression of Lymphatic Marker mRNA after ADS C-Derived EVs Treatment 
     Materials and Methods 
     qRT-PCR 
     To assess the effect of ADSC-EVs on LECs, confluent LECs in 24-well plates were treated with 500 μl of EBM-2 containing PBS, rVEGF-C(10 ng/ml), or ADSC-EVs(10 μg/ml) for 12 or 24 hours. Total RNA was extracted from LECs cultured in each condition using a QIAzol and the miRNeasy Mini Kit (Qiagen, Holden, Germany) according to the manufacturer&#39;s protocols. For qRT-PCR analysis, complementary DNA was generated from 1 μg of total RNA using a High Capacity cDNA Reverse Transcription Kit (Applied Biosystems). Real-Time PCR system was subsequently performed in triplicate with a 1:15 dilution of cDNA using TaqMan Gene Expression Assays (Applied Biosystems) on a StepOne Real-Time PCR System (Applied Biosystems). Each Assay ID (Thermo Fisher Scientific) is LYVE-1 (Hs00272659_m1), VEFF-R3 (Hs01047677_m1), Prox1 (Hs00896293_m1), and podoplanin (Hs00366766_m1). All mRNA quantification data from cultured cells were normalized to the expression of glyceraldehydes 3-phosphate dehydrogenase (GAPDH). 
     Results 
     Lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1), vascular endothelial growth factor receptor-3 (VEGFR-3), prospero homeobox 1 (Prox1), and podoplanin are well known lymphatic markers in LECs. Therefore, expression levels of mRNA of LYVE-1, VEGFR-3, Prox1, and podoplanin in LECs were examined under conditions of PBS, VEGF-C, or ADSC-EVs treatment. The mRNA expression of LYVE-1 was slightly higher in both 12 and 24 hours in the ADSC-EVs treatment group compared to the PBS group. The VEGF-C group showed a similar trend to the ADSC-EVs treatment group. As for VEGFR-3 mRNA expression, it was almost four times higher in 12 hours and twice higher in 24 hours in the ADSC-EVs treatment group than the PBS treatment group. The VEGF-C treated group showed five to six times higher expression than the PBS treated group in 12 hours. The mRNA expression of Prox1 showed no significant differences in 12 hours, however, it was almost 2.5 times higher in the ADSC-EVs treatment group than PBS treatment group, after 24 hours. VEGF-C treatment group showed little expression changes compared to the ADSC-EVs treatment group. The expression levels of Podoplanin were increased 1.5 times higher in ADSC-EVs treatment group than PBS treatment group in 12 hours; however, it showed no significant differences in 24 hours. VEGF-C group showed the same trend as the ADSC-EVs treatment group. See  FIGS. 3A-3H . 
     Example 4 
     ADSC-Derived EV Improved Hindlimbs Appearance in a Lymphedema Mouse Model 
     Materials and Methods 
     Lymphedema Mouse Model, Treatment and Edema Assessment 
     Lymphedema was established in the bilateral hindlimbs of 10-week-old female C57BL/6J mice (CLEA Japan, Inc., Tokyo, Japan) as previously described (Iwasaki, et al.,  Plast Reconstr Surg.,  139(1):67e-78e (2017)). They were handled according to the guidelines established for animal care at the center and the protocol was approved by the Institutional Animal Care and Use Committee of Jichi Medical University. After gas anesthesia, the mice were subjected to x-ray irradiation in the bilateral inguinal region at 10 Gy in a single dose twice prior to the surgery. The radiation was emitted from an x-ray machine (MX-160Labo, mediXtec, Japan). One week later, mice were subjected to circumferential incision in the inguinal region to the muscle layer. The subiliac and popliteal lymph nodes and the lymphatic vessels entwined around the sciatic veins were removed. To block the superficial lymphatic system, a 3-mm-wide silicone splint was placed in the inguinal wound and then fixed to the skin and underlying muscle using interrupted 6-0 nylon sutures (Bear medic, Tokyo, Japan). This silicon splint placement prevented wound contraction and desiccation. As a therapeutic intervention, injection of PBS (25 μ1) or ADSC-EVs (˜40 μg) or HEK293-EVs (˜40 μg) as a thin layer to the whole leg area was performed in postoperative day 7 and 14. Hindlimb circumference measurements were performed by unblind reviewers at a point 6 mm proximal to the heel at various time points. 
     Fluorescence lymphography was used to compare the lymphatic structures in swollen hindlimbs treated with PBS or ADSC-EVs every 2 weeks postoperatively using a near-infrared fluorescence camera system (FLUORO, Toa Kogaku, Japan). To examine the thickness of soft tissue postoperatively, we used computed tomography (CT) imaging over 4 weeks postoperatively. 
     Edema Assessment 
     To measure the extent of postsurgical edema, hindlimb circumference measurements were performed at a point 6 mm proximal to the heel. In addition, to evaluate the state of lymphedema and the effect of injection of ADSC-EVs, hindlimb circumferences were determined at various time points. Fluorescence lymphography was used to compare lymphatic structures in swollen hindlimbs treated PBS or ADSC-EVs every 2 weeks postoperatively. The animals were clipped and residual hair was removed with a depilatory cream before imaging. Anesthetized mice with isoflurane were placed on a warming pad. A 5-μl volume of a 1-mg/ml solution of indocyanine green (Sigma-Aldrich, St. Louis, Mo.) dissolved in distilled water was injected subcutaneously into the dorsal aspect of both paws using a 26-gauge needle. Fluorescence images were acquired 15 mintes after indocyanine green injection using a near-infrared fluorescence camera system (FLUORO, Toa Kogaku, Japan). Computed tomography (CT) imaging was used to examine the thickness of soft tissue postoperatively, over 4 weeks. Anesthesized mice with isoflurane were placed inside the CT. After the CT images were acquired, the thickness of the soft tissue (length from bone to skin) was measured. 
     Results 
     All limbs demonstrated consistent enlargement after radiation, surgical removal of lymph nodes and lymphatic vessels and silicone fixation ( FIG. 4A ). Although the limb volumes did not return to preoperative levels in these animals, treatment with ADSC-EVs was highly effective in decreasing gross leg swelling (65.1% +−4.5%) as compared with controls (79.4% +−5.1%) ( FIG. 4B ). 
     To assess lymphatic function, indocyanine green lymphography of the hindlimb was performed. Using indocyanine green lymphography 4 weeks after the final treatment, clearance of accumulated lymphatic fluid and a clear linear sign of lymphatic channels in ADSC-EVs-treated legs. In contrast, control legs demonstrated pooling of ICG in the whole area and no transport across the zone of surgery ( FIG. 4C ). This finding was confirmed using computed tomographic (CT) images. The thickness of soft tissue area, which was defined between bone and skin, markedly decreased in the ADSC-EVs treated group compared with the controls, indicating that the excess amount of lymphatic fluid was drained into the venous system. 
     Example 5 
     ADSC-Derived EVs Promoted Angiogenesis and Lymphangiogenesis in the Affected Limb 
     Materials and Methods 
     Immunohistochemistry 
     The mice were sacrificed 4 weeks after the final injection of PBS or ADSC-EVs. Skin sections (5 μm) were generated from 6 mm distal to the inguinal wound and 6 mm proximal to the ankle. The tissues were stained with Hoechst 33342 (Dojindo, Tokyo, Japan), lymphatic vessel endothelial receptor-1 (LYVE-1) polyclonal antibodies (bs-1311R-A555, Bioss Antibodies, USA), anti-CD31 polyclonal antibodies (bs-0468R-A647, Bioss Antibodies, USA), SMAD3 polyclonal antibodies (bs-2225R-A488, Bioss Antibodies, USA), and collagen 1 polyclonal antibodies (bs-10423R-A488, Bioss Antibodies, USA). Stained slides were examined under a fluorescence microscope (Keyence, Osaka, Japan). Fluorescence images were captured at 40×magnification in 5 random fields. For quantification, each positive area was measured using NIH ImageJ software by unblind reviewers. 
     Results 
     To determine the lymphangiogenic and angiogenic effects of ADSC-EVs in lymphedema leg models in which drainage of lymphatic fluid is obstructed surgically, histologic changes of affected limbs were evaluated. Immunohistochemical analysis of LYVE-1+ vessel counts in the hindlimb tissues of experimental and control legs demonstrated that injection of ADSC-EVs group modestly but significantly increased the total number of capillary lymphatic vessels (25.3%+−5.2%, P&lt;0.05) compared to PBS group and HEK293-EVs group (16.1%+−2.8%, 16.9%+−3.2% respectively) ( FIG. 5A ). Similarly, analysis of cross-sections obtained from the hindlimb in the mouse models demonstrated increase in the number of CD31+ endothelial cells in the hindlimb tissues treated with ADSC-EVs (12.3%+−3.9%) as compared with PBS or HEK293-EVs (7.7%+−3.3%, 10.2%+−2.2%, respectively) ( FIG. 5B ). Consistent with the increased number of vascular and lymphatic endothelial cells, an increase of the number of vessels which expressed both CD31+ and LYVE-1+ in tissue samples harvested from the ADSC-EVs(5.2%+−1.1%, P&lt;0.05) group was noted compared to controls (2.1%+−0.2%, 2.3%+−0.3%) ( FIG. 5C ). These overlapping regions may be the newly connected bypass between lymphatics to capillary vessels, indicating that interstitial transport capacity is greatly increased after ADSC-EVs treatment. 
     Example 6 
     ADSC-Derived EVs Decreased Fibrosis After Lymphatic Injury 
     Because the degree of fibrosis in lymphedema patients correlates with the severity of disease, several markers of fibrosis were analyzed in hindlimbs to understand the effects of ADSC-EVs treatment. ADSC-EVs significantly decreased subdermal type I collagen deposition (32.5%+−4.4%, P&lt;0.05) as compared with PBS group (45.8%+−5.1%) and HEK293-EVs group (42.2%+−4.5%) ( FIG. 6A ). ADSC-EVs treatment significantly decreased expression of phosphorylated SMAD3 (pSMAD-3) (17.1%+−9.2%, P&lt;0.05), which is a downstream signaling molecule of pro-fibrotic growth factor, compared to PBS group(36.5%+−14.2%) and HEK293-EVs group (23.3%+−10.4%) ( FIG. 6B ). Taken together, these findings show that ADSC-EVs mitigate the formation of fibrotic response in the setting of lymphedema. 
     Example 7 
     ADSC-Derived EVs Contain Several miRNAs Targeting MDM2, HIF1a, and PHB 
     Materials and Methods 
     miRNA Expression Profiling by Real-Time PCR Arrays 
     Total RNA, including miRNA, was extracted from ADSC-EVs and HEK293-EVs using Trizol reagent (Invitrogen Life Technologies, Carlsbad, CA, USA) according to the manufacturer&#39;s instruction. From each sample, 15 μl of miRNAs were reverse-transcribed using a miRCURY LNA RT Kit (Qiagen, Netherlands). The cDNA was mixed with 2× PCR master mix (Qiagen, Netherlands). All the real-time PCR-based experiments using a 
     LightCycler 480 Instrument 2 according to the manufacturer&#39;s instructions. For normalization, miRNA expressions were compared between ADSC-EVs and HEK293-EVs. The cycle threshold (2-AACt) method was utilized to calculate the fold change. 
     mRNA Expression Profiling in LECs by Microarray 
     To perform a mRNA microarray, 5×10e4 LECs were seeded into 24-well plates and 12 h later EVs from ADSC or PBS as control were added to LECs. Total mRNAs were extracted from LECs 12 h after treatment with the EVs and PBS. For each hybridization, 0.60 μg of Cy3 labeled cRNA were fragmented, and hybridized at 65 degrees for 17 hours to an Agilent SurePrint G3 Human GE v3 8×60K Microarray (Design ID: 072363). After washing, microarrays were scanned using an Agilent DNA microarray scanner. 
     Intensity values of each scanned feature were quantified using Agilent feature extraction software version 11.5.1.1, which performs background subtractions. Only features that were flagged as no errors (Detected flags) were used and features that were not positive, not significant, not uniform, not above background, saturated, and population outliers (Not Detected and Compromised flags) were excluded. Normalization was performed using Agilent GeneSpring software version 14.9.1 (per chip:normalization to Quantile). 
     Results 
     To determine how ADSC-EVs regulate miRNAs associated with various processes, using a real-time RT-PCR-based miRNA array, the expression patterns of 752 different miRNAs were next profiled in ADSC-EVs. The expression patterns of the same miRNAs in HEK293 cells derived EVs were also analyzed as controls. The miRNA PCR array revealed that 396 differentially expressed miRNAs in ADSC-EVs compared to HEK293-EVs. miRNAs expressed over double fold in log2 ratio are shown in Table 1. 
     Bioinformatic analysis (Ingenuity Pathway Analysis, IPA), was performed to evaluate the implications of the altered miRNA expression. IPA revealed that the major networks which incorporate the predicted targets, include functions such as ‘Organismal injury and abnormalities’, ‘Reproductive system disease’, ‘Inflammatory disease’, and ‘Inflammatory response’. 
     The cargo of EVs, which includes miRNAs, mRNAs, proteins and other biological components, is transferred from the donor to recipient cells and influences the cellular phenotypes. Therefore, which genes are altered in the LECs treated with the ADSC-EVs or PBS as control were investigated. 
     Total RNA was extracted from treated LECs, and microarray analyses were performed. Results show that differentially expressed genes among treatments with ADSC-EVs and control PBS, there were clear differences in gene expression in the LECs. 
     A bioinformatic analysis was conducted using IPA for the relation between highly expressed miRNAs in ADSC-EVs and genes which showed expression changes in LECs after addition of ADSC-EVs ( FIG. 7 , Table 2). As a result, eight miRNAs such as miR-199a-3p, miR-145-5p, miR-143-3p, miR-377-3p, miR-100-3p, miR-29a-3p, miR-495-3p, and miR-29c-3p were found to target and regulate the expression of mouse double minute 2 homolog (MDM2), which contributed to the stability of hypoxiainducible factor-1 alpha (HIF1a) and resulted in angiogenesis and lymphangiogenesis in LECs.. MDM2 is a cellular oncoprotein encoded by a gene located on chromosome 12q13-14. MDM2 can suppress p53, a cancer suppressor gene. Inhibition of MDM2 has anti-inflammatory effects and may lead to treatment of autoimmune diseases and cancer. Previous studies have shown that MDM2 negatively regulates the stability of hypoxia-inducible factor-1 alpha (HIF1a) in protein level. This investigation found that the expression level of HIF1a was elevated in LECs after addition of ADSC-EVs. HIF1a is a transcription factor that controls the cellular response to hypoxia. HIF1a promotes transcription of various proteins such as VEGF, Erythropoietin (EPO) and glucose transporters, and also plays a key role in lymphangiogenesis and angiogenesis. The result indicates that the reduced expression of MDM2 caused by miR-199a-3p, miR-145-5p, miR-143-3p, miR-377-3p, miR-100-3p, miR-29a-3p, miR-495-3p, and miR-29c-3p in ADSC-EVs lead to elevation of HIF1a in LECs, which leads to angiogenesis and lymphangiogenesis. 
     Another target of four miRNAs such as miR-658, miR-493-3p, miR-184, and miR-27a-3p in ADSC-EVs was found to be Prohibitin (PHB). PHB is a membrane protein which regulates a variety of biological processes such as apoptosis, cell cycle, signal transduction and cellular senescence. The downregulation of PHB can lead to angiogenesis (Qian, et al.,  CNS Neurosci Ther.,  19(10):804-12 (2013)). 
     
       
         
           
               
             
               
                 TABLE 1 
               
             
            
               
                   
               
               
                 miRNAs expressed over double fold in log2 ratio 
               
            
           
           
               
               
               
               
            
               
                 SEQ ID # 
                 miRname 
                 Log2 Ratio 
                 target sequence 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                   1 
                 hsa-miR-548c-3p 
                 13.1573 
                 CAAAAAUCUCAAUUACUUUUGC 
               
               
                   
               
               
                   2 
                 hsa-miR-214-5p 
                 12.4406 
                 UGCCUGUCUACACUUGCUGUGC 
               
               
                   
               
               
                   3 
                 hsa-miR-372-3p 
                 12.4273 
                 AAAGUGCUGCGACAUUUGAGCGU 
               
               
                   
               
               
                   4 
                 hsa-miR-206 
                 12.0473 
                 UGGAAUGUAAGGAAGUGUGUGG 
               
               
                   
               
               
                   5 
                 hsa-miR-337-5p 
                 11.2373 
                 GAACGGCUUCAUACAGGAGUU 
               
               
                   
               
               
                   6 
                 hsa-miR-199a-5p 
                 10.2440 
                 CCCAGUGUUCAGACUACCUGUUC 
               
               
                   
               
               
                   7 
                 hsa-miR-214-3p 
                 9.2040 
                 ACAGCAGGCACAGACAGGCAGU 
               
               
                   
               
               
                   8 
                 hsa-miR-199a-3p 
                 8.9240 
                 ACAGUAGUCUGCACAUUGGUUA 
               
               
                   
               
               
                   9 
                 hsa-miR-370-3p 
                 8.4440 
                 GCCUGCUGGGGUGGAACCUGGU 
               
               
                   
               
               
                  10 
                 hsa-miR-100-5p 
                 7.2940 
                 AACCCGUAGAUCCGAACUUGUG 
               
               
                   
               
               
                  11 
                 hsa-miR-145-5p 
                 7.2440 
                 GUCCAGUUUUCCCAGGAAUCCCU 
               
               
                   
               
               
                  12 
                 hsa-miR-199b-5p 
                 7.1940 
                 CCCAGUGUUUAGACUAUCUGUUC 
               
               
                   
               
               
                  13 
                 hsa-miR-143-3p 
                 7.1640 
                 UGAGAUGAAGCACUGUAGCUC 
               
               
                   
               
               
                  14 
                 hsa-miR-888-5p 
                 7.0240 
                 UACUCAAAAAGCUGUCAGUCA 
               
               
                   
               
               
                  15 
                 hsa-let-71-3p 
                 6.8206 
                 CUGCGCAAGCUACUGCCUUGCU 
               
               
                   
               
               
                  16 
                 hsa-let-7 a-2-3p 
                 6.7106 
                 CUGUACAGCCUCCUAGCUUUCC 
               
               
                   
               
               
                  17 
                 hsa-miR-137 
                 6.6740 
                 UUAUUGCUUAAGAAUACGCGUAG 
               
               
                   
               
               
                  18 
                 hsa-miR-337-3p 
                 6.4640 
                 CUCCUAUAUGAUGCCUUUCUUC 
               
               
                   
               
               
                  19 
                 hsa-miR-145-3p 
                 6.3173 
                 GGAUUCCUGGAAAUACUGUUCU 
               
               
                   
               
               
                  20 
                 hsa-miR-133b 
                 6.2640 
                 UUUGGUCCCCUUCAACCAGCUA 
               
               
                   
               
               
                  21 
                 hsa-miR-208b-3p 
                 5.7706 
                 AUAAGACGAACAAAAGGUUUGU 
               
               
                   
               
               
                  22 
                 hsa-miR-493-5p 
                 5.6873 
                 UUGUACAUGGUAGGCUUUCAUU 
               
               
                   
               
               
                  23 
                 hsa-let-7b-3p 
                 5.6073 
                 CUAUACAACCUACUGCCUUCCC 
               
               
                   
               
               
                  24 
                 hsa-let-71-5p 
                 5.6040 
                 UGAGGUAGUAGUUUGUGCUGUU 
               
               
                   
               
               
                  25 
                 hsa-miR-143-5p 
                 5.5773 
                 GGUGCAGUGCUGCAUCUCUGGU 
               
               
                   
               
               
                  26 
                 hsa-miR-675-5p 
                 5.4673 
                 UGGUGCGGAGAGGGCCCACAGUG 
               
               
                   
               
               
                  27 
                 hsa-miR-136-3p 
                 5.4473 
                 CAUCAUCGUCUCAAAUGAGUCU 
               
               
                   
               
               
                  28 
                 hsa-miR-133a-3p 
                 5.3740 
                 UUUGGUCCCCUUCAACCAGCUG 
               
               
                   
               
               
                  29 
                 hsa-miR-377-5p 
                 5.2406 
                 AGAGGUUGCCCUUGGUGAAUUC 
               
               
                   
               
               
                  30 
                 hsa-let-7b-5p 
                 5.2340 
                 UGAGGUAGUAGGUUGUGUGGUU 
               
               
                   
               
               
                  31 
                 hsa-miR-377-3p 
                 5.1840 
                 AUCACACAAAGGCAACUUUUGU 
               
               
                   
               
               
                  32 
                 hsa-miR-136-5p 
                 5.1840 
                 ACUCCAUUUGUUUUGAUGAUGGA 
               
               
                   
               
               
                  33 
                 hsa-miR-125b-1-3p 
                 5.0673 
                 ACGGGUUAGGCUCUUGGGAGCU 
               
               
                   
               
               
                  34 
                 hsa-miR-100-3p 
                 5.040584 
                 CAAGCUUGUAUCUAUAGGUAUG 
               
               
                   
               
               
                  35 
                 hsa-miR-125b-5p 
                 4.9640 
                 UCCCUGAGACCCUAACUUGUGA 
               
               
                   
               
               
                  36 
                 hsa-miR-127-3p 
                 4.8840 
                 UCGGAUCCGUCUGAGCUUGGCU 
               
               
                   
               
               
                  37 
                 hsa-miR-29a-5p 
                 4.8673 
                 ACUGAUUUCUUUUGGUGUUCAG 
               
               
                   
               
               
                  38 
                 hsa-miR-135a-3p 
                 4.8106 
                 UAUAGGGAUUGGAGCCGUGGCG 
               
               
                   
               
               
                  39 
                 hsa-miR-539-5p 
                 4.7340 
                 GGAGAAAUUAUCCUUGGUGUGU 
               
               
                   
               
               
                  40 
                 hsa-miR-424-3p 
                 4.5773 
                 CAAAACGUGAGGCGCUGCUAU 
               
               
                   
               
               
                  41 
                 hsa-miR-369-5p 
                 4.5540 
                 AGAUCGACCGUGUUAUAUUCGC 
               
               
                   
               
               
                  42 
                 hsa-miR-376a-3p 
                 4.2940 
                 AUCAUAGAGGAAAAUCCACGU 
               
               
                   
               
               
                  43 
                 hsa-miR-410-3p 
                 4.2340 
                 AAUAUAACACAGAUGGCCUGU 
               
               
                   
               
               
                  44 
                 hsa-miR-376c-3p 
                 4.1940 
                 AACAUAGAGGAAAUUCCACGU 
               
               
                   
               
               
                  45 
                 hsa-miR-154-5p 
                 4.1840 
                 UAGGUUAUCCGUGUUGCCUUCG 
               
               
                   
               
               
                  46 
                 hsa-miR-409-3p 
                 4.1740 
                 GAAUGUUGCUCGGUGAACCCCU 
               
               
                   
               
               
                  47 
                 hsa-miR-146a-5p 
                 4.0940 
                 UGAGAACUGAAUUCCAUGGGUU 
               
               
                   
               
               
                  48 
                 hsa-miR-29a-3p 
                 4.0640 
                 UAGCACCAUCUGAAAUCGGUUA 
               
               
                   
               
               
                  49 
                 hsa-miR-127-5p 
                 3.9640 
                 CUGAAGCUCAGAGGGCUCUGAU 
               
               
                   
               
               
                  50 
                 hsa-miR-382-5p 
                 3.9540 
                 GAAGUUGUUCGUGGUGGAUUCG 
               
               
                   
               
               
                  51 
                 hsa-miR-409-5p 
                 3.9273 
                 AGGUUACCCGAGCAACUUUGCAU 
               
               
                   
               
               
                  52 
                 hsa-miR-424-5p 
                 3.8540 
                 CAGCAGCAAUUCAUGUUUUGAA 
               
               
                   
               
               
                  53 
                 hsa-let-7f-1-3p 
                 3.7873 
                 CUAUACAAUCUAUUGCCUUCCC 
               
               
                   
               
               
                  54 
                 hsa-miR-485-3p 
                 3.7140 
                 GUCAUACACGGCUCUCCUCUCU 
               
               
                   
               
               
                  55 
                 hsa-miR-369-3p 
                 3.6273 
                 AAUAAUACAUGGUUGAUCUUU 
               
               
                   
               
               
                  56 
                 hsa-miR-376a-5p 
                 3.6173 
                 GUAGAUUCUCCUUCUAUGAGUA 
               
               
                   
               
               
                  57 
                 hsa-miR-1 
                 3.5940 
                 UGGAAUGUAAAGAAGUAUGUAU 
               
               
                   
               
               
                  58 
                 hsa-let-7d-3p 
                 3.5340 
                 CUAUACGACCUGCUGCCUUUCU 
               
               
                   
               
               
                  59 
                 hsa-miR-382-3p 
                 3.4973 
                 AAUCAUUCACGGACAACACUU 
               
               
                   
               
               
                  60 
                 hsa-miR-668-3p 
                 3.4840 
                 UGUCACUCGGCUCGGCCCACUAC 
               
               
                   
               
               
                  61 
                 hsa-miR-487b-3p 
                 3.4340 
                 AAUCGUACAGGGUCAUCCACUU 
               
               
                   
               
               
                  62 
                 hsa-miR-411-5p 
                 3.4240 
                 UAGUAGACCGUAUAGCGUACG 
               
               
                   
               
               
                  63 
                 hsa-miR-544a 
                 3.4073 
                 AUUCUGCAUUUUUAGCAAGUUC 
               
               
                   
               
               
                  64 
                 hsa-miR-889-3p 
                 3.4073 
                 UUAAUAUCGGACAACCAUUGU 
               
               
                   
               
               
                  65 
                 hsa-miR-329-3p 
                 3.2840 
                 AACACACCUGGUUAACCUCUUU 
               
               
                   
               
               
                  66 
                 hsa-miR-503-5p 
                 3.2540 
                 UAGCAGCGGGAACAGUUCUGCAG 
               
               
                   
               
               
                  67 
                 hsa-miR-30c-1-3p 
                 3.2473 
                 CUGGGAGAGGGUUGUUUACUCC 
               
               
                   
               
               
                  68 
                 hsa-miR-22-5p 
                 3.1840 
                 AGUUCUUCAGUGGCAAGCUUUA 
               
               
                   
               
               
                  69 
                 hsa-miR-658 
                 3.1473 
                 GGCGGAGGGAAGUAGGUCCGUUGGU 
               
               
                   
               
               
                  70 
                 hsa-miR-1185-5p 
                 3.1773 
                 AGAGGAUACCCUUUGUAUGUU 
               
               
                   
               
               
                  71 
                 hsa-miR-495-3p 
                 3.1240 
                 AAACAAACAUGGUGCACUUCUU 
               
               
                   
               
               
                  72 
                 hsa-miR-432-5p 
                 3.1040 
                 UCUUGGAGUAGGUCAUUGGGUGG 
               
               
                   
               
               
                  73 
                 hsa-miR-379-5p 
                 3.1040 
                 UGGUAGACUAUGGAACGUAGG 
               
               
                   
               
               
                  74 
                 hsa-miR-22-3p 
                 3.0640 
                 AAGCUGCCAGUUGAAGAACUGU 
               
               
                   
               
               
                  75 
                 hsa-miR-654-3p 
                 3.0373 
                 UAUGUCUGCUGACCAUCACCUU 
               
               
                   
               
               
                  76 
                 hsa-miR-365b-5p 
                 3.0273 
                 AGGGACUUUCAGGGGCAGCUGU 
               
               
                   
               
               
                  77 
                 hsa-miR-493-3p 
                 2.9740 
                 UGAAGGUCUACUGUGUGCCAGG 
               
               
                   
               
               
                  78 
                 hsa-miR-134-5p 
                 2.9340 
                 UGUGACUGGUUGACCAGAGGGG 
               
               
                   
               
               
                  79 
                 hsa-miR-154-3p 
                 2.8740 
                 AAUCAUACACGGUUGACCUAUU 
               
               
                   
               
               
                  80 
                 hsa-miR-411-3p 
                 2.8273 
                 UAUGUAACACGGUCCACUAACC 
               
               
                   
               
               
                  81 
                 hsa-miR-335-3p 
                 2.7773 
                 UUUUUCAUUAUUGCUCCUGACC 
               
               
                   
               
               
                  82 
                 hsa-miR-543 
                 2.7573 
                 AAACAUUCGCGGUGCACUUCUU 
               
               
                   
               
               
                  83 
                 hsa-miR-381-3p 
                 2.6940 
                 UAUACAAGGGCAAGCUCUCUGU 
               
               
                   
               
               
                  84 
                 hsa-miR-574-3p 
                 2.6640 
                 CACGCUCAUGCACACACCCACA 
               
               
                   
               
               
                  85 
                 hsa-miR-379-3p 
                 2.6173 
                 UAUGUAACAUGGUCCACUAACU 
               
               
                   
               
               
                  86 
                 hsa-miR-21-5p 
                 2.6040 
                 UAGCUUAUCAGACUGAUGUUGA 
               
               
                   
               
               
                  87 
                 hsa-miR-654-5p 
                 2.6040 
                 UGGUGGGCCGCAGAACAUGUGC 
               
               
                   
               
               
                  88 
                 hsa-miR-605-5p 
                 2.5573 
                 UAAAUCCCAUGGUGCCUUCUCCU 
               
               
                   
               
               
                  89 
                 hsa-miR-29c-3p 
                 2.5440 
                 UAGCACCAUUUGAAAUCGGUUA 
               
               
                   
               
               
                  90 
                 hsa-miR-184 
                 2.5040 
                 UGGACGGAGAACUGAUAAGGGU 
               
               
                   
               
               
                  91 
                 hsa-miR-193a-5p 
                 2.4940 
                 UGGGUCUUUGCGGGCGAGAUGA 
               
               
                   
               
               
                  92 
                 hsa-miR-541-5p 
                 2.4773 
                 AAAGGAUUCUGCUGUCGGUCCCACU 
               
               
                   
               
               
                  93 
                 hs a-miR-27 a-3p 
                 2.4740 
                 UUCACAGUGGCUAAGUUCCGC 
               
               
                   
               
               
                  94 
                 hsa-miR-494-3p 
                 2.4240 
                 UGAAACAUACACGGGAAACCUC 
               
               
                   
               
               
                  95 
                 hsa-miR-376b-3p 
                 2.4140 
                 AUCAUAGAGGAAAAUCCAUGUU 
               
               
                   
               
               
                  96 
                 hsa-miR- 144-5p 
                 2.3673 
                 GGAUAUCAUCAUAUACUGUAAG 
               
               
                   
               
               
                  97 
                 hsa-miR-1244 
                 2.3473 
                 AAGUAGUUGGUUUGUAUGAGAUGGUU 
               
               
                   
               
               
                  98 
                 hsa-miR-323a-3p 
                 2.3440 
                 CACAUUACACGGUCGACCUCU 
               
               
                   
               
               
                  99 
                 hsa-miR-335-5p 
                 2.2940 
                 UCAAGAGCAAUAACGAAAAAUGU 
               
               
                   
               
               
                 100 
                 hsa-miR-21-3p 
                 2.2640 
                 CAACACCAGUCGAUGGGCUGU 
               
               
                   
               
               
                 101 
                 hs a-let-7 d-5p 
                 2.0940 
                 AGAGGUAGUAGGUUGCAUAGUU 
               
               
                   
               
               
                 102 
                 hsa-miR-487 a-3p 
                 2.0773 
                 AAUCAUACAGGGACAUCCAGUU 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 2 
               
             
            
               
                   
               
               
                 Differentially expressed genes in LEC for treatment of EVs or PBS 
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                   
                 Fold 
                 Log fold 
               
               
                   
                   
                 Expression 
                 change 
                 change 
               
               
                   
                   
                 compared to 
                 (ADSC EV 
                 (ADSC EV 
               
               
                 GeneSymbol 
                 GeneName 
                 control 
                 vs PBS) 
                 vs PBS) 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 FOXB1 
                 forkhead box B1 
                 up 
                 19.85 
                 4.31 
               
               
                 WWC2-AS1 
                 WWC2 antisense RNA 1 
                 up 
                 19.66 
                 4.30 
               
               
                 CFAP61 
                 cilia and flagella associated 
                 up 
                 14.71 
                 3.88 
               
               
                   
                 protein 61 
               
               
                 CHST10 
                 carbohydrate sulfotransferase 10 
                 up 
                 13.57 
                 3.76 
               
               
                 HRK 
                 harakiri, BCL2 interacting 
                 up 
                 12.81 
                 3.68 
               
               
                   
                 protein 
               
               
                 KLK8 
                 kallikrein-related peptidase 8 
                 up 
                 12.43 
                 3.64 
               
               
                 MYHAS 
                 myosin heavy chain gene cluster 
                 up 
                 12.15 
                 3.60 
               
               
                   
                 antisense RNA 
               
               
                 HTR4 
                 5-hydroxytryptamine (serotonin) 
                 up 
                 11.96 
                 3.58 
               
               
                   
                 receptor 4, G protein-coupled 
               
               
                 ANAPC1 
                 anaphase promoting complex 
                 up 
                 10.64 
                 3.41 
               
               
                   
                 subunit 1 
               
               
                 LYPD6B 
                 LY6/PLAUR domain containing 
                 up 
                 9.75 
                 3.29 
               
               
                   
                 6B 
               
               
                 LIG3 
                 ligase III, DNA, ATP-dependent 
                 up 
                 9.75 
                 3.29 
               
               
                 TSPEAR 
                 thrombospondin-type laminin G 
                 up 
                 9.41 
                 3.23 
               
               
                   
                 domain and EAR repeats 
               
               
                 OR6A2 
                 olfactory receptor, family 6, 
                 up 
                 9.28 
                 3.21 
               
               
                   
                 subfamily A, member 2 
               
               
                 GRIN2C 
                 glutamate receptor, ionotropic, 
                 up 
                 9.21 
                 3.20 
               
               
                   
                 N-methyl D-aspartate 2C 
               
               
                 GPR141 
                 G protein-coupled receptor 141 
                 up 
                 9.11 
                 3.19 
               
               
                 MUC7 
                 mucin 7, secreted 
                 up 
                 8.51 
                 3.09 
               
               
                 ATXN8OS 
                 ATXN8 opposite strand (non- 
                 up 
                 8.14 
                 3.03 
               
               
                   
                 protein coding) 
               
               
                 KLK13 
                 kallikrein-related peptidase 13 
                 up 
                 8.05 
                 3.01 
               
               
                 ADAMTS8 
                 ADAM metallopeptidase with 
                 up 
                 8.01 
                 3.00 
               
               
                   
                 thrombospondin type 1 motif, 8 
               
               
                 ILF2 
                 interleukin enhancer binding 
                 up 
                 7.59 
                 2.92 
               
               
                   
                 factor 2 
               
               
                 RMDN2-AS1 
                 RMDN2 antisense RNA 1 
                 up 
                 7.48 
                 2.90 
               
               
                 CD300C 
                 CD300c molecule 
                 up 
                 7.44 
                 2.90 
               
               
                 LRP1 
                 low density lipoprotein receptor- 
                 up 
                 7.25 
                 2.86 
               
               
                   
                 related protein 1 
               
               
                 CCDC13 
                 coiled-coil domain containing 13 
                 up 
                 6.77 
                 2.76 
               
               
                 POLM 
                 polymerase (DNA directed), mu 
                 up 
                 6.40 
                 2.68 
               
               
                 GPER1 
                 G protein-coupled estrogen 
                 up 
                 6.37 
                 2.67 
               
               
                   
                 receptor 1 
               
               
                 SNORD77 
                 small nucleolar RNA, C/D box 
                 up 
                 6.14 
                 2.62 
               
               
                   
                 77 
               
               
                 SDC2 
                 syndecan 2 
                 up 
                 5.78 
                 2.53 
               
               
                 KRTAP22-1 
                 keratin associated protein 22-1 
                 up 
                 5.76 
                 2.53 
               
               
                 SLITRK2 
                 SLIT and NTRK-like family, 
                 up 
                 5.72 
                 2.52 
               
               
                   
                 member 2 
               
               
                 LRRC7 
                 leucine rich repeat containing 7 
                 up 
                 5.51 
                 2.46 
               
               
                 CHRNA2 
                 cholinergic receptor, nicotinic, 
                 up 
                 5.40 
                 2.43 
               
               
                   
                 alpha 2 (neuronal) 
               
               
                 SEPT3 
                 septin 3 
                 up 
                 5.22 
                 2.39 
               
               
                 TSRM 
                 zinc finger domain-related 
                 up 
                 5.14 
                 2.36 
               
               
                   
                 protein TSRM 
               
               
                 TRAPPC2 
                 trafficking protein particle 
                 up 
                 5.03 
                 2.33 
               
               
                   
                 complex 2 
               
               
                 WDR5B 
                 WD repeat domain 5B 
                 up 
                 4.97 
                 2.31 
               
               
                 PPP1R26-AS1 
                 PPP1R26 antisense RNA 1 
                 up 
                 4.79 
                 2.26 
               
               
                 SAMD13 
                 sterile alpha motif domain 
                 up 
                 4.71 
                 2.23 
               
               
                   
                 containing 13 
               
               
                 RNF180 
                 ring finger protein 180 
                 up 
                 4.69 
                 2.23 
               
               
                 ZNF888 
                 zinc finger protein 888 
                 up 
                 4.39 
                 2.14 
               
               
                 PTCD2 
                 pentatricopeptide repeat domain 
                 up 
                 4.38 
                 2.13 
               
               
                   
                 2 
               
               
                 TM4SF5 
                 transmembrane 4 L six family 
                 up 
                 4.37 
                 2.13 
               
               
                   
                 member 5 
               
               
                 PHGR1 
                 proline/histidine/glycine-rich 1 
                 up 
                 4.33 
                 2.11 
               
               
                 LRRC10 
                 leucine rich repeat containing 10 
                 up 
                 4.27 
                 2.09 
               
               
                 COL27A1 
                 collagen, type XXVII, alpha 1 
                 up 
                 4.26 
                 2.09 
               
               
                 XKR7 
                 XK, Kell blood group complex 
                 up 
                 4.16 
                 2.06 
               
               
                   
                 subunit-related family, member 7 
               
               
                 NOS1AP 
                 nitric oxide synthase 1 (neuronal) 
                 up 
                 4.11 
                 2.04 
               
               
                   
                 adaptor protein 
               
               
                 ITLN2 
                 intelectin 2 
                 up 
                 4.11 
                 2.04 
               
               
                 PTCD2 
                 pentatricopeptide repeat domain 
                 up 
                 4.05 
                 2.02 
               
               
                   
                 2 
               
               
                 KRT8 
                 keratin 8, type II 
                 up 
                 3.92 
                 1.97 
               
               
                 FAM150B 
                 family with sequence similarity 
                 up 
                 3.85 
                 1.94 
               
               
                   
                 150, member B 
               
               
                 SNORD19B 
                 small nucleolar RNA, C/D box 
                 up 
                 3.83 
                 1.94 
               
               
                   
                 19B 
               
               
                 MEGF10 
                 multiple EGF-like-domains 10 
                 up 
                 3.76 
                 1.91 
               
               
                 LRIG2 
                 leucine-rich repeats and 
                 up 
                 3.67 
                 1.87 
               
               
                   
                 immunoglobulin-like domains 2 
               
               
                 LRIT1 
                 leucine-rich repeat, 
                 up 
                 3.62 
                 1.86 
               
               
                   
                 immunoglobulin-like and 
               
               
                   
                 transmembrane domains 1 
               
               
                 OR1L8 
                 olfactory receptor, family 1, 
                 up 
                 3.56 
                 1.83 
               
               
                   
                 subfamily L, member 8 
               
               
                 CBX3P2 
                 chromobox homolog 3 
                 up 
                 3.54 
                 1.83 
               
               
                   
                 pseudogene 2 
               
               
                 RAB11FIP4 
                 RAB11 family interacting 
                 up 
                 3.50 
                 1.81 
               
               
                   
                 protein 4 (class II) 
               
               
                 FSD1L 
                 fibronectin type III and SPRY 
                 up 
                 3.48 
                 1.80 
               
               
                   
                 domain containing 1-like 
               
               
                 ROR1 
                 receptor tyrosine kinase-like 
                 up 
                 3.45 
                 1.79 
               
               
                   
                 orphan receptor 1 
               
               
                 CCDC17 
                 coiled-coil domain containing 17 
                 up 
                 3.42 
                 1.78 
               
               
                 SLC7A14 
                 solute carrier family 7, member 
                 up 
                 3.42 
                 1.77 
               
               
                   
                 14 
               
               
                 KCNS1 
                 potassium voltage-gated channel, 
                 up 
                 3.41 
                 1.77 
               
               
                   
                 modifier subfamily S, member 1 
               
               
                 PRY2 
                 PTPN13-like, Y-linked 2 
                 up 
                 3.40 
                 1.76 
               
               
                 TLCD2 
                 TLC domain containing 2 
                 up 
                 3.40 
                 1.76 
               
               
                 CEP85L 
                 centrosomal protein 85 kDa-like 
                 up 
                 3.34 
                 1.74 
               
               
                 CDC42EP3 
                 CDC42 effector protein (Rho 
                 up 
                 3.29 
                 1.72 
               
               
                   
                 GTPase binding) 3 
               
               
                 PVRL3-AS1 
                 PVRL3 antisense RNA 1 
                 up 
                 3.24 
                 1.69 
               
               
                 NT5DC4 
                 5′-nucleotidase domain 
                 up 
                 3.23 
                 1.69 
               
               
                   
                 containing 4 
               
               
                 CNOT4 
                 CCR4-NOT transcription 
                 up 
                 3.18 
                 1.67 
               
               
                   
                 complex, subunit 4 
               
               
                 POFUT1 
                 protein O-fucosyltransferase 1 
                 up 
                 3.17 
                 1.67 
               
               
                 PTGER4P2- 
                 PTGER4P2-CDK2AP2P2 
                 up 
                 3.17 
                 1.66 
               
               
                 CDK2AP2P2 
                 readthrough transcribed 
               
               
                   
                 pseudogene 
               
               
                 NCOA2 
                 nuclear receptor coactivator 2 
                 up 
                 3.13 
                 1.65 
               
               
                 L2HGDH 
                 L-2-hydroxyglutarate 
                 up 
                 3.12 
                 1.64 
               
               
                   
                 dehydrogenase 
               
               
                 SIRPG 
                 signal-regulatory protein gamma 
                 up 
                 3.10 
                 1.63 
               
               
                 MID1 
                 midline 1 
                 up 
                 3.10 
                 1.63 
               
               
                 WDR55 
                 WD repeat domain 55 
                 up 
                 3.10 
                 1.63 
               
               
                 GATA6 
                 GATA binding protein 6 
                 up 
                 3.09 
                 1.63 
               
               
                 RNF216 
                 ring finger protein 216 
                 up 
                 3.06 
                 1.61 
               
               
                 CGN 
                 cingulin 
                 up 
                 3.04 
                 1.60 
               
               
                 SERPINI1 
                 serpin peptidase inhibitor, clade I 
                 up 
                 3.01 
                 1.59 
               
               
                   
                 (neuroserpin), member 1 
               
               
                 LCP1 
                 lymphocyte cytosolic protein 1 
                 up 
                 3.01 
                 1.59 
               
               
                   
                 (L-plastin) 
               
               
                 NR2F2-AS1 
                 NR2F2 antisense RNA 1 
                 up 
                 3.00 
                 1.59 
               
               
                 FLJ43315 
                 asparagine synthetase 
                 up 
                 2.99 
                 1.58 
               
               
                   
                 pseudogene 
               
               
                 AADAC 
                 arylacetamide deacetylase 
                 up 
                 2.99 
                 1.58 
               
               
                 NDUFA6-AS1 
                 NDUFA6 antisense RNA 1 (head 
                 up 
                 2.98 
                 1.57 
               
               
                   
                 to head) 
               
               
                 PMFBP1 
                 polyamine modulated factor 1 
                 up 
                 2.96 
                 1.57 
               
               
                   
                 binding protein 1 
               
               
                 RBM23 
                 RNA binding motif protein 23 
                 up 
                 2.96 
                 1.57 
               
               
                 CALML3-AS1 
                 CALML3 antisense RNA 1 
                 up 
                 2.96 
                 1.56 
               
               
                 CSRNP3 
                 cysteine-serine-rich nuclear 
                 up 
                 2.96 
                 1.56 
               
               
                   
                 protein 3 
               
               
                 PCDHGC3 
                 protocadherin gamma subfamily 
                 up 
                 2.95 
                 1.56 
               
               
                   
                 C, 3 
               
               
                 CCDC110 
                 coiled-coil domain containing 
                 up 
                 2.95 
                 1.56 
               
               
                   
                 110 
               
               
                 SYDE2 
                 synapse defective 1, Rho 
                 up 
                 2.94 
                 1.56 
               
               
                   
                 GTPase, homolog 2 ( C. elegans ) 
               
               
                 LRRC7 
                 leucine rich repeat containing 7 
                 up 
                 2.92 
                 1.54 
               
               
                 SYNGAP1 
                 synaptic Ras GTPase activating 
                 up 
                 2.91 
                 1.54 
               
               
                   
                 protein 1 
               
               
                 MMP25 
                 matrix metallopeptidase 25 
                 up 
                 2.88 
                 1.52 
               
               
                 PLXDC1 
                 plexin domain containing 1 
                 up 
                 2.87 
                 1.52 
               
               
                 SPIN3 
                 spindlin family, member 3 
                 up 
                 2.87 
                 1.52 
               
               
                 SP2-AS1 
                 SP2 antisense RNA 1 
                 up 
                 2.87 
                 1.52 
               
               
                 FAM178B 
                 family with sequence similarity 
                 up 
                 2.87 
                 1.52 
               
               
                   
                 178, member B 
               
               
                 FAM104B 
                 family with sequence similarity 
                 up 
                 2.86 
                 1.52 
               
               
                   
                 104, member B 
               
               
                 SSU72 
                 SSU72 RNA polymerase II CTD 
                 up 
                 2.86 
                 1.52 
               
               
                   
                 phosphatase homolog 
               
               
                   
                 ( S. cerevisiae ) 
               
               
                 CALY 
                 calcyon neuron-specific vesicular 
                 up 
                 2.86 
                 1.52 
               
               
                   
                 protein 
               
               
                 ACVR1B 
                 activin A receptor, type IB 
                 up 
                 2.84 
                 1.51 
               
               
                 KCTD21-AS1 
                 KCTD21 antisense RNA 1 
                 up 
                 2.84 
                 1.50 
               
               
                 GPR173 
                 G protein-coupled receptor 173 
                 up 
                 2.84 
                 1.50 
               
               
                 LBX1-AS1 
                 LBX1 antisense RNA 1 (head to 
                 up 
                 2.83 
                 1.50 
               
               
                   
                 head) 
               
               
                 SPDYE3 
                 speedy/RINGO cell cycle 
                 up 
                 2.82 
                 1.50 
               
               
                   
                 regulator family member E3 
               
               
                 CHRDL1 
                 chordin-like 1 
                 up 
                 2.82 
                 1.50 
               
               
                 CDK20 
                 cyclin-dependent kinase 20 
                 up 
                 2.80 
                 1.49 
               
               
                 SLC51B 
                 solute carrier family 51, beta 
                 up 
                 2.80 
                 1.48 
               
               
                   
                 subunit 
               
               
                 ZBED3-AS1 
                 ZBED3 antisense RNA 1 
                 up 
                 2.78 
                 1.48 
               
               
                 ETV4 
                 ets variant 4 
                 up 
                 2.77 
                 1.47 
               
               
                 KLHL15 
                 kelch-like family member 15 
                 up 
                 2.75 
                 1.46 
               
               
                 TCTE3 
                 t-complex-associated-testis- 
                 up 
                 2.75 
                 1.46 
               
               
                   
                 expressed 3 
               
               
                 MAGEA2B 
                 melanoma antigen family A, 2B 
                 up 
                 2.74 
                 1.46 
               
               
                 SP2-AS1 
                 SP2 antisense RNA 1 
                 up 
                 2.74 
                 1.46 
               
               
                 RPAP2 
                 RNA polymerase II associated 
                 up 
                 2.74 
                 1.45 
               
               
                   
                 protein 2 
               
               
                 ITPK1-AS1 
                 ITPK1 antisense RNA 1 
                 up 
                 2.73 
                 1.45 
               
               
                 ATP1A1-AS1 
                 ATP1A1 antisense RNA 1 
                 up 
                 2.73 
                 1.45 
               
               
                 C2orf15 
                 chromosome 2 open reading 
                 up 
                 2.72 
                 1.44 
               
               
                   
                 frame 15 
               
               
                 UBE2E4P 
                 ubiquitin-conjugating enzyme 
                 up 
                 2.71 
                 1.44 
               
               
                   
                 E2E 4 pseudogene 
               
               
                 OCLN 
                 occludin 
                 up 
                 2.71 
                 1.44 
               
               
                 CHAC1 
                 ChaC glutathione-specific 
                 up 
                 2.70 
                 1.43 
               
               
                   
                 gamma-glutamylcyclotransferase 
               
               
                   
                 1 
               
               
                 SPOPL 
                 speckle-type POZ protein-like 
                 up 
                 2.69 
                 1.43 
               
               
                 TMEM200B 
                 transmembrane protein 200B 
                 up 
                 2.68 
                 1.42 
               
               
                 TMEM184B 
                 transmembrane protein 184B 
                 up 
                 2.68 
                 1.42 
               
               
                 GLTPD2 
                 glycolipid transfer protein 
                 up 
                 2.65 
                 1.41 
               
               
                   
                 domain containing 2 
               
               
                 OBP2B 
                 odorant binding protein 2B 
                 up 
                 2.64 
                 1.40 
               
               
                 CYP4Z1 
                 cytochrome P450, family 4, 
                 up 
                 2.64 
                 1.40 
               
               
                   
                 subfamily Z, polypeptide 1 
               
               
                 CASC11 
                 cancer susceptibility candidate 
                 up 
                 2.63 
                 1.40 
               
               
                   
                 11 (non-protein coding) 
               
               
                 AHCTF1 
                 AT hook containing transcription 
                 up 
                 2.63 
                 1.40 
               
               
                   
                 factor 1 
               
               
                 PFKFB2 
                 6-phosphofructo-2- 
                 up 
                 2.62 
                 1.39 
               
               
                   
                 kinase/fructose-2,6- 
               
               
                   
                 biphosphatase 2 
               
               
                 FAM154B 
                 family with sequence similarity 
                 up 
                 2.62 
                 1.39 
               
               
                   
                 154, member B 
               
               
                 SUPT3H 
                 suppressor of Ty 3 homolog 
                 up 
                 2.61 
                 1.39 
               
               
                   
                 ( S. cerevisiae ) 
               
               
                 BAALC 
                 brain and acute leukemia, 
                 up 
                 2.61 
                 1.38 
               
               
                   
                 cytoplasmic 
               
               
                 RIBC1 
                 RIB43A domain with coiled- 
                 up 
                 2.59 
                 1.37 
               
               
                   
                 coils 1 
               
               
                 ZNF354C 
                 zinc finger protein 354C 
                 up 
                 2.59 
                 1.37 
               
               
                 DIS3E2 
                 DIS3 like 3′-5′ exoribonuclease 2 
                 up 
                 2.58 
                 1.37 
               
               
                 HIST1H2AA 
                 histone cluster 1, H2aa 
                 up 
                 2.55 
                 1.35 
               
               
                 MYCNOS 
                 MYCN opposite strand 
                 up 
                 2.55 
                 1.35 
               
               
                 PLA2G2D 
                 phospholipase A2, group IID 
                 up 
                 2.54 
                 1.34 
               
               
                 ANKRD20A19P 
                 ankyrin repeat domain 20 family, 
                 up 
                 2.52 
                 1.33 
               
               
                   
                 member A19, pseudogene 
               
               
                 SSPO 
                 SCO-spondin 
                 up 
                 2.52 
                 1.33 
               
               
                 ARMCX4 
                 armadillo repeat containing, X- 
                 up 
                 2.51 
                 1.33 
               
               
                   
                 linked 4 
               
               
                 SNAR-G1 
                 small ILF3/NF90-associated 
                 up 
                 2.51 
                 1.33 
               
               
                   
                 RNA G1 
               
               
                 MYLK3 
                 myosin light chain kinase 3 
                 up 
                 2.50 
                 1.32 
               
               
                 NPAS3 
                 neuronal PAS domain protein 3 
                 up 
                 2.49 
                 1.31 
               
               
                 HS1BP3-IT1 
                 HS1BP3 intronic transcript 1 
                 up 
                 2.49 
                 1.31 
               
               
                   
                 (non-protein coding) 
               
               
                 DTX3 
                 deltex 3, E3 ubiquitin ligase 
                 up 
                 2.48 
                 1.31 
               
               
                 PGM5P4-AS1 
                 PGM5P4 antisense RNA 1 
                 up 
                 2.46 
                 1.30 
               
               
                 SNORA14B 
                 small nucleolar RNA, H/ACA 
                 up 
                 2.46 
                 1.30 
               
               
                   
                 box 14B 
               
               
                 SLC39A2 
                 solute carrier family 39 (zinc 
                 up 
                 2.46 
                 1.30 
               
               
                   
                 transporter), member 2 
               
               
                 ZNF287 
                 zinc finger protein 287 
                 up 
                 2.45 
                 1.29 
               
               
                 GS1-259H13.2 
                 transmembrane protein 225-like 
                 up 
                 2.45 
                 1.29 
               
               
                 MRGPRG 
                 MAS-related GPR, member G 
                 up 
                 2.45 
                 1.29 
               
               
                 LRRC17 
                 leucine rich repeat containing 17 
                 up 
                 2.45 
                 1.29 
               
               
                 LANCL2 
                 LanC lantibiotic synthetase 
                 up 
                 2.44 
                 1.29 
               
               
                   
                 component C-like 2 (bacterial) 
               
               
                 ANKEF1 
                 ankyrin repeat and EF-hand 
                 up 
                 2.44 
                 1.29 
               
               
                   
                 domain containing 1 
               
               
                 PBX2 
                 pre-B-cell leukemia homeobox 2 
                 up 
                 2.43 
                 1.28 
               
               
                 CCDC121 
                 coiled-coil domain containing 
                 up 
                 2.42 
                 1.28 
               
               
                   
                 121 
               
               
                 CRYBA2 
                 crystallin, beta A2 
                 up 
                 2.42 
                 1.28 
               
               
                 TMPO-AS1 
                 TMPO antisense RNA 1 
                 up 
                 2.42 
                 1.28 
               
               
                 ACSBG1 
                 acyl-CoA synthetase bubblegum 
                 up 
                 2.41 
                 1.27 
               
               
                   
                 family member 1 
               
               
                 MR1 
                 major histocompatibility 
                 up 
                 2.41 
                 1.27 
               
               
                   
                 complex, class I-related 
               
               
                 PPIP5K1 
                 diphosphoinositol 
                 up 
                 2.41 
                 1.27 
               
               
                   
                 pentakisphosphate kinase 1 
               
               
                 TRIM6 
                 tripartite motif containing 6 
                 up 
                 2.41 
                 1.27 
               
               
                 CCDC121 
                 coiled-coil domain containing 
                 up 
                 2.41 
                 1.27 
               
               
                   
                 121 
               
               
                 TUSC5 
                 tumor suppressor candidate 5 
                 up 
                 2.40 
                 1.26 
               
               
                 XPO5 
                 exportin 5 
                 up 
                 2.39 
                 1.26 
               
               
                 SPDYE8P 
                 speedy/RINGO cell cycle 
                 up 
                 2.39 
                 1.26 
               
               
                   
                 regulator family member E8, 
               
               
                   
                 pseudogene 
               
               
                 CSRNP3 
                 cysteine-serine-rich nuclear 
                 up 
                 2.39 
                 1.26 
               
               
                   
                 protein 3 
               
               
                 RBM33 
                 RNA binding motif protein 33 
                 up 
                 2.37 
                 1.25 
               
               
                 B3GNT7 
                 UDP-GlcNAc:betaGal beta-1,3- 
                 up 
                 2.37 
                 1.24 
               
               
                   
                 N-acetylglucosaminyltransferase 
               
               
                   
                 7 
               
               
                 NME9 
                 NME/NM23 family member 9 
                 up 
                 2.36 
                 1.24 
               
               
                 CYP4V2 
                 cytochrome P450, family 4, 
                 up 
                 2.36 
                 1.24 
               
               
                   
                 subfamily V, polypeptide 2 
               
               
                 CDSN 
                 corneodesmosin 
                 up 
                 2.36 
                 1.24 
               
               
                 COL20A1 
                 collagen, type XX, alpha 1 
                 up 
                 2.36 
                 1.24 
               
               
                 ZNF192P1 
                 zinc finger protein 192 
                 up 
                 2.35 
                 1.23 
               
               
                   
                 pseudogene 1 
               
               
                 HERC6 
                 HECT and RLD domain 
                 up 
                 2.35 
                 1.23 
               
               
                   
                 containing E3 ubiquitin protein 
               
               
                   
                 ligase family member 6 
               
               
                 SNORD25 
                 small nucleolar RNA, C/D box 
                 up 
                 2.35 
                 1.23 
               
               
                   
                 25 
               
               
                 FRS3 
                 fibroblast growth factor receptor 
                 up 
                 2.35 
                 1.23 
               
               
                   
                 substrate 3 
               
               
                 RGS5 
                 regulator of G-protein signaling 
                 up 
                 2.33 
                 1.22 
               
               
                   
                 5 
               
               
                 RMST 
                 rhabdomyosarcoma 2 associated 
                 up 
                 2.33 
                 1.22 
               
               
                   
                 transcript (non-protein coding) 
               
               
                 GAS2L2 
                 growth arrest-specific 2 like 2 
                 up 
                 2.33 
                 1.22 
               
               
                 SGPP1 
                 sphingosine-1-phosphate 
                 up 
                 2.32 
                 1.21 
               
               
                   
                 phosphatase 1 
               
               
                 NIPSNAP3B 
                 nipsnap homolog 3B ( C. elegans ) 
                 up 
                 2.31 
                 1.21 
               
               
                 TAB3 
                 TGF-beta activated kinase 
                 up 
                 2.31 
                 1.21 
               
               
                   
                 1/MAP3K7 binding protein 3 
               
               
                 RASSF10 
                 Ras association (RalGDS/AF-6) 
                 up 
                 2.31 
                 1.21 
               
               
                   
                 domain family (N-terminal) 
               
               
                   
                 member 10 
               
               
                 PPIL6 
                 peptidylprolyl isomerase 
                 up 
                 2.31 
                 1.20 
               
               
                   
                 (cyclophilin)-like 6 
               
               
                 MPP4 
                 membrane protein, palmitoylated 
                 up 
                 2.30 
                 1.20 
               
               
                   
                 4 (MAGUK p55 subfamily 
               
               
                   
                 member 4) 
               
               
                 ZNF37A 
                 zinc finger protein 37A 
                 up 
                 2.30 
                 1.20 
               
               
                 ATG10 
                 autophagy related 10 
                 up 
                 2.29 
                 1.20 
               
               
                 FPR3 
                 formyl peptide receptor 3 
                 up 
                 2.28 
                 1.19 
               
               
                 PDP2 
                 pyruvate dehyrogenase 
                 up 
                 2.27 
                 1.18 
               
               
                   
                 phosphatase catalytic subunit 2 
               
               
                 ZFAND4 
                 zinc finger, AN1-type domain 4 
                 up 
                 2.27 
                 1.18 
               
               
                 RHOT1 
                 ras homolog family member T1 
                 up 
                 2.26 
                 1.18 
               
               
                 NKAPP1 
                 NFKB activating protein 
                 up 
                 2.26 
                 1.18 
               
               
                   
                 pseudogene 1 
               
               
                 OPTC 
                 opticin 
                 up 
                 2.26 
                 1.18 
               
               
                 FLT4 
                 fms-related tyrosine kinase 4 
                 up 
                 2.26 
                 1.18 
               
               
                 OR2AG1 
                 olfactory receptor, family 2, 
                 up 
                 2.26 
                 1.17 
               
               
                   
                 subfamily AG, member 1 
               
               
                   
                 (gene/pseudogene) 
               
               
                 C9orf72 
                 chromosome 9 open reading 
                 up 
                 2.25 
                 1.17 
               
               
                   
                 frame 72 
               
               
                 PPIP5K1 
                 diphosphoinositol 
                 up 
                 2.25 
                 1.17 
               
               
                   
                 pentakisphosphate kinase 1 
               
               
                 ASIC3 
                 acid sensing (proton gated) ion 
                 up 
                 2.24 
                 1.17 
               
               
                   
                 channel 3 
               
               
                 PIK3CD 
                 phosphatidylinositol-4,5- 
                 up 
                 2.24 
                 1.16 
               
               
                   
                 bisphosphate 3-kinase, catalytic 
               
               
                   
                 subunit delta 
               
               
                 FAAHP1 
                 fatty acid amide hydrolase 
                 up 
                 2.24 
                 1.16 
               
               
                   
                 pseudogene 1 
               
               
                 ZNF641 
                 zinc finger protein 641 
                 up 
                 2.24 
                 1.16 
               
               
                 SCN1B 
                 sodium channel, voltage gated, 
                 up 
                 2.24 
                 1.16 
               
               
                   
                 type I beta subunit 
               
               
                 FCN1 
                 ficolin (collagen/fibrinogen 
                 up 
                 2.22 
                 1.15 
               
               
                   
                 domain containing) 1 
               
               
                 HIST1H4G 
                 histone cluster 1, H4g 
                 up 
                 2.22 
                 1.15 
               
               
                 CEBPA 
                 CCAAT/enhancer binding 
                 up 
                 2.22 
                 1.15 
               
               
                   
                 protein (C/EBP), alpha 
               
               
                 ANKRD20A8P 
                 ankyrin repeat domain 20 family, 
                 up 
                 2.22 
                 1.15 
               
               
                   
                 member A8, pseudogene 
               
               
                 MLYCD 
                 malonyl-CoA decarboxylase 
                 up 
                 2.22 
                 1.15 
               
               
                 C5orf56 
                 chromosome 5 open reading 
                 up 
                 2.22 
                 1.15 
               
               
                   
                 frame 56 
               
               
                 HERC6 
                 HECT and RLD domain 
                 up 
                 2.22 
                 1.15 
               
               
                   
                 containing E3 ubiquitin protein 
               
               
                   
                 ligase family member 6 
               
               
                 PRKACB 
                 protein kinase, cAMP-dependent, 
                 up 
                 2.22 
                 1.15 
               
               
                   
                 catalytic, beta 
               
               
                 SRP14-AS1 
                 SRP14 antisense RNA1 (head to 
                 up 
                 2.22 
                 1.15 
               
               
                   
                 head) 
               
               
                 C1RL 
                 complement component 1, r 
                 up 
                 2.21 
                 1.15 
               
               
                   
                 subcomponent-like 
               
               
                 EMR2 
                 egf-like module containing, 
                 up 
                 2.21 
                 1.15 
               
               
                   
                 mucin-like, hormone receptor- 
               
               
                   
                 like 2 
               
               
                 ATPAF1 
                 ATP synthase mitochondrial F1 
                 up 
                 2.21 
                 1.15 
               
               
                   
                 complex assembly factor 1 
               
               
                 ZNF222 
                 zinc finger protein 222 
                 up 
                 2.20 
                 1.14 
               
               
                 SCGB1B2P 
                 secretoglobin, family 1B, 
                 up 
                 2.20 
                 1.14 
               
               
                   
                 member 2, pseudogene 
               
               
                 CCDC113 
                 coiled-coil domain containing 
                 up 
                 2.20 
                 1.14 
               
               
                   
                 113 
               
               
                 C15orf48 
                 chromosome 15 open reading 
                 up 
                 2.20 
                 1.14 
               
               
                   
                 frame 48 
               
               
                 ETV3 
                 ets variant 3 
                 up 
                 2.20 
                 1.14 
               
               
                 SCP2 
                 sterol carrier protein 2 
                 up 
                 2.20 
                 1.14 
               
               
                 MGC34796 
                 sepiapterin reductase (7,8- 
                 up 
                 2.19 
                 1.13 
               
               
                   
                 dihydrobiopterin:NADP+ 
               
               
                   
                 oxidoreductase) pseudogene 
               
               
                 RAP1GAP 
                 RAP1 GTPase activating protein 
                 up 
                 2.19 
                 1.13 
               
               
                 MAGOHB 
                 mago-nashi homolog B 
                 up 
                 2.19 
                 1.13 
               
               
                   
                 ( Drosophila ) 
               
               
                 ZBED4 
                 zinc finger, BED-type containing 
                 up 
                 2.19 
                 1.13 
               
               
                   
                 4 
               
               
                 FAM98B 
                 family with sequence similarity 
                 up 
                 2.18 
                 1.13 
               
               
                   
                 98, member B 
               
               
                 EPHB1 
                 EPH receptor B1 
                 up 
                 2.18 
                 1.13 
               
               
                 POM121 
                 POM121 transmembrane 
                 up 
                 2.18 
                 1.12 
               
               
                   
                 nucleoporin 
               
               
                 SNX21 
                 sorting nexin family member 21 
                 up 
                 2.18 
                 1.12 
               
               
                 OR6Y1 
                 olfactory receptor, family 6, 
                 up 
                 2.18 
                 1.12 
               
               
                   
                 subfamily Y, member 1 
               
               
                 LMBRD2 
                 LMBR1 domain containing 2 
                 up 
                 2.18 
                 1.12 
               
               
                 FLVCR2 
                 feline leukemia virus subgroup C 
                 up 
                 2.17 
                 1.12 
               
               
                   
                 cellular receptor family, member 
               
               
                   
                 2 
               
               
                 LRRC15 
                 leucine rich repeat containing 15 
                 up 
                 2.17 
                 1.12 
               
               
                 TGFBR3 
                 transforming growth factor, beta 
                 up 
                 2.17 
                 1.12 
               
               
                   
                 receptor III 
               
               
                 NXT2 
                 nuclear transport factor 2-like 
                 up 
                 2.17 
                 1.11 
               
               
                   
                 export factor 2 
               
               
                 HCRP1 
                 hepatocellular carcinoma-related 
                 up 
                 2.17 
                 1.11 
               
               
                   
                 HCRP1 
               
               
                 HEPACAM 
                 hepatic and glial cell adhesion 
                 up 
                 2.16 
                 1.11 
               
               
                   
                 molecule 
               
               
                 OBSCN 
                 obscurin, cytoskeletal calmodulin 
                 up 
                 2.16 
                 1.11 
               
               
                   
                 and titin-interacting RhoGEF 
               
               
                 SQSTM1 
                 sequestosome 1 
                 up 
                 2.16 
                 1.11 
               
               
                 LGALS8 
                 lectin, galactoside-binding, 
                 up 
                 2.15 
                 1.11 
               
               
                   
                 soluble, 8 
               
               
                 RHBDL1 
                 rhomboid, veinlet-like 1 
                 up 
                 2.15 
                 1.11 
               
               
                   
                 ( Drosophila ) 
               
               
                 KIF21B 
                 kinesin family member 21B 
                 up 
                 2.15 
                 1.10 
               
               
                 CEP170 
                 centrosomal protein 170 kDa 
                 up 
                 2.15 
                 1.10 
               
               
                 CYP51A1-AS1 
                 CYP51A1 antisense RNA 1 
                 up 
                 2.15 
                 1.10 
               
               
                 LIMD1 
                 LIM domains containing 1 
                 up 
                 2.15 
                 1.10 
               
               
                 ANKRD53 
                 ankyrin repeat domain 53 
                 up 
                 2.14 
                 1.10 
               
               
                 INGX 
                 inhibitor of growth family, X- 
                 up 
                 2.14 
                 1.10 
               
               
                   
                 linked, pseudogene 
               
               
                 CDH15 
                 cadherin 15, type 1, M-cadherin 
                 up 
                 2.14 
                 1.10 
               
               
                   
                 (myotubule) 
               
               
                 APCS 
                 amyloid P component, serum 
                 up 
                 2.14 
                 1.10 
               
               
                 SYN2 
                 synapsin II 
                 up 
                 2.14 
                 1.10 
               
               
                 SMKR1 
                 small lysine-rich protein 1 
                 up 
                 2.13 
                 1.09 
               
               
                 KCNK7 
                 potassium channel, two pore 
                 up 
                 2.13 
                 1.09 
               
               
                   
                 domain subfamily K, member 7 
               
               
                 TEX28 
                 testis expressed 28 
                 up 
                 2.13 
                 1.09 
               
               
                 DYNLRB2 
                 dynein, light chain, roadblock- 
                 up 
                 2.13 
                 1.09 
               
               
                   
                 type 2 
               
               
                 CNNM2 
                 cyclin and CBS domain divalent 
                 up 
                 2.13 
                 1.09 
               
               
                   
                 metal cation transport mediator 2 
               
               
                 RSG1 
                 REM2 and RAB-like small 
                 up 
                 2.13 
                 1.09 
               
               
                   
                 GTPase 1 
               
               
                 FRMD8 
                 FERM domain containing 8 
                 up 
                 2.13 
                 1.09 
               
               
                 STK33 
                 serine/threonine kinase 33 
                 up 
                 2.13 
                 1.09 
               
               
                 OPLAH 
                 5-oxoprolinase (ATP- 
                 up 
                 2.13 
                 1.09 
               
               
                   
                 hydrolysing) 
               
               
                 EFHC1 
                 EF-hand domain (C-terminal) 
                 up 
                 2.13 
                 1.09 
               
               
                   
                 containing 1 
               
               
                 C16orf47 
                 chromosome 16 open reading 
                 up 
                 2.13 
                 1.09 
               
               
                   
                 frame 47 
               
               
                 GSTO2 
                 glutathione S-transferase omega 
                 up 
                 2.12 
                 1.09 
               
               
                   
                 2 
               
               
                 PTPLB 
                 protein tyrosine phosphatase-like 
                 up 
                 2.12 
                 1.09 
               
               
                   
                 (proline instead of catalytic 
               
               
                   
                 arginine), member b 
               
               
                 HIST1H2APS1 
                 histone cluster 1, H2a, 
                 up 
                 2.12 
                 1.09 
               
               
                   
                 pseudogene 1 
               
               
                 OR10W1 
                 olfactory receptor, family 10, 
                 up 
                 2.12 
                 1.08 
               
               
                   
                 subfamily W, member 1 
               
               
                 PEX5 
                 peroxisomal biogenesis factor 5 
                 up 
                 2.12 
                 1.08 
               
               
                 ASS1 
                 argininosuccinate synthase 1 
                 up 
                 2.11 
                 1.08 
               
               
                 LRRC16B 
                 leucine rich repeat containing 
                 up 
                 2.11 
                 1.08 
               
               
                   
                 16B 
               
               
                 CWC15 
                 CWC15 spliceosome-associated 
                 up 
                 2.11 
                 1.08 
               
               
                   
                 protein 
               
               
                 SUV420H1 
                 suppressor of variegation 4-20 
                 up 
                 2.11 
                 1.08 
               
               
                   
                 homolog 1 ( Drosophila ) 
               
               
                 TET3 
                 tet methylcytosine dioxygenase 3 
                 up 
                 2.11 
                 1.07 
               
               
                 POLD2 
                 polymerase (DNA directed), 
                 up 
                 2.10 
                 1.07 
               
               
                   
                 delta 2, accessory subunit 
               
               
                 CCDC169 
                 coiled-coil domain containing 
                 up 
                 2.10 
                 1.07 
               
               
                   
                 169 
               
               
                 QRICH2 
                 glutamine rich 2 
                 up 
                 2.10 
                 1.07 
               
               
                 SLC9A5 
                 solute carrier family 9, subfamily 
                 up 
                 2.10 
                 1.07 
               
               
                   
                 A (NHE5, cation proton 
               
               
                   
                 antiporter 5), member 5 
               
               
                 CFAP69 
                 cilia and flagella associated 
                 up 
                 2.10 
                 1.07 
               
               
                   
                 protein 69 
               
               
                 HSF1 
                 heat shock transcription factor 1 
                 up 
                 2.09 
                 1.07 
               
               
                 ACTB 
                 actin, beta 
                 up 
                 2.09 
                 1.06 
               
               
                 MYPOP 
                 Myb-related transcription factor, 
                 up 
                 2.09 
                 1.06 
               
               
                   
                 partner of profilin 
               
               
                 HOXA11 
                 homeobox A11 
                 up 
                 2.08 
                 1.06 
               
               
                 GNG12-AS1 
                 GNG12 antisense RNA 1 
                 up 
                 2.08 
                 1.06 
               
               
                 TAPBP 
                 TAP binding protein (tapasin) 
                 up 
                 2.08 
                 1.06 
               
               
                 DLEU2L 
                 deleted in lymphocytic leukemia 
                 up 
                 2.08 
                 1.06 
               
               
                   
                 2-like 
               
               
                 TESC 
                 tescalcin 
                 up 
                 2.08 
                 1.06 
               
               
                 ITGA8 
                 integrin, alpha 8 
                 up 
                 2.08 
                 1.05 
               
               
                 PPARGC1B 
                 peroxisome proliferator-activated 
                 up 
                 2.08 
                 1.05 
               
               
                   
                 receptor gamma, coactivator 1 
               
               
                   
                 beta 
               
               
                 FAM26E 
                 family with sequence similarity 
                 up 
                 2.08 
                 1.05 
               
               
                   
                 26, member E 
               
               
                 OR2A9P 
                 olfactory receptor, family 2, 
                 up 
                 2.08 
                 1.05 
               
               
                   
                 subfamily A, member 9 
               
               
                   
                 pseudogene 
               
               
                 WHAMMP3 
                 WAS protein homolog associated 
                 up 
                 2.07 
                 1.05 
               
               
                   
                 with actin, golgi membranes and 
               
               
                   
                 microtubules pseudogene 3 
               
               
                 BTNL10 
                 butyrophilin-like 10 
                 up 
                 2.07 
                 1.05 
               
               
                 PADI6 
                 peptidyl arginine deiminase, type 
                 up 
                 2.07 
                 1.05 
               
               
                   
                 VI 
               
               
                 TMEM86A 
                 transmembrane protein 86A 
                 up 
                 2.06 
                 1.04 
               
               
                 PARK2 
                 parkin RBR E3 ubiquitin protein 
                 up 
                 2.06 
                 1.04 
               
               
                   
                 ligase 
               
               
                 MAPT 
                 microtubule-associated protein 
                 up 
                 2.06 
                 1.04 
               
               
                   
                 tau 
               
               
                 MAGEC1 
                 melanoma antigen family C, 1 
                 up 
                 2.06 
                 1.04 
               
               
                 PYCR1 
                 pyrroline-5-carboxylate 
                 up 
                 2.06 
                 1.04 
               
               
                   
                 reductase 1 
               
               
                 CTBP1-AS2 
                 CTBP1 antisense RNA 2 (head 
                 up 
                 2.05 
                 1.04 
               
               
                   
                 to head) 
               
               
                 GGCT 
                 gamma-glutamylcyclotransferase 
                 up 
                 2.05 
                 1.04 
               
               
                 FAM25A 
                 family with sequence similarity 
                 up 
                 2.05 
                 1.03 
               
               
                   
                 25, member A 
               
               
                 PTCH1 
                 patched 1 
                 up 
                 2.05 
                 1.03 
               
               
                 ALDH5A1 
                 aldehyde dehydrogenase 5 
                 up 
                 2.05 
                 1.03 
               
               
                   
                 family, member A1 
               
               
                 CCDC68 
                 coiled-coil domain containing 68 
                 up 
                 2.05 
                 1.03 
               
               
                 DAB2IP 
                 DAB2 interacting protein 
                 up 
                 2.05 
                 1.03 
               
               
                 ATAD3C 
                 ATPase family, AAA domain 
                 up 
                 2.05 
                 1.03 
               
               
                   
                 containing 3C 
               
               
                 MARVELD2 
                 MARVEL domain containing 2 
                 up 
                 2.04 
                 1.03 
               
               
                 KLC4 
                 kinesin light chain 4 
                 up 
                 2.04 
                 1.03 
               
               
                 SREK1IP1 
                 SREK1-interacting protein 1 
                 up 
                 2.04 
                 1.03 
               
               
                 CHAC2 
                 ChaC, cation transport regulator 
                 up 
                 2.04 
                 1.03 
               
               
                   
                 homolog 2 ( E. coli ) 
               
               
                 HDHD3 
                 haloacid dehalogenase-like 
                 up 
                 2.04 
                 1.03 
               
               
                   
                 hydrolase domain containing 3 
               
               
                 ELMO2 
                 engulfment and cell motility 2 
                 up 
                 2.04 
                 1.03 
               
               
                 TMSB4Y 
                 thymosin beta 4, Y-linked 
                 up 
                 2.04 
                 1.03 
               
               
                 CYP2B6 
                 cytochrome P450, family 2, 
                 up 
                 2.03 
                 1.02 
               
               
                   
                 subfamily B, polypeptide 6 
               
               
                 EDNRB 
                 endothelin receptor type B 
                 up 
                 2.03 
                 1.02 
               
               
                 ZNF660 
                 zinc finger protein 660 
                 up 
                 2.03 
                 1.02 
               
               
                 SLC2A3 
                 solute carrier family 2 (facilitated 
                 up 
                 2.03 
                 1.02 
               
               
                   
                 glucose transporter), member 3 
               
               
                 ZNF749 
                 zinc finger protein 749 
                 up 
                 2.03 
                 1.02 
               
               
                 C2orf27A 
                 chromosome 2 open reading 
                 up 
                 2.03 
                 1.02 
               
               
                   
                 frame 27A 
               
               
                 STRADA 
                 STE20-related kinase adaptor 
                 up 
                 2.03 
                 1.02 
               
               
                   
                 alpha 
               
               
                 MIR99AHG 
                 mir-99a-let-7c cluster host gene 
                 up 
                 2.03 
                 1.02 
               
               
                   
                 (non-protein coding) 
               
               
                 ZNF671 
                 zinc finger protein 671 
                 up 
                 2.02 
                 1.02 
               
               
                 CEP44 
                 centrosomal protein 44 kDa 
                 up 
                 2.02 
                 1.02 
               
               
                 VPS9D1-AS1 
                 VPS9D1 antisense RNA 1 
                 up 
                 2.02 
                 1.01 
               
               
                 SMAD1-AS1 
                 SMAD1 antisense RNA 1 
                 up 
                 2.02 
                 1.01 
               
               
                 UBE3C 
                 ubiquitin protein ligase E3C 
                 up 
                 2.02 
                 1.01 
               
               
                 PRSS33 
                 protease, serine, 33 
                 up 
                 2.02 
                 1.01 
               
               
                 TTC28 
                 tetratricopeptide repeat domain 
                 up 
                 2.02 
                 1.01 
               
               
                   
                 28 
               
               
                 RASGRF1 
                 Ras protein-specific guanine 
                 up 
                 2.01 
                 1.01 
               
               
                   
                 nucleotide-releasing factor 1 
               
               
                 CFH 
                 complement factor H 
                 up 
                 2.01 
                 1.01 
               
               
                 GNG12-AS1 
                 GNG12 antisense RNA 1 
                 up 
                 2.01 
                 1.01 
               
               
                 GUSBP5 
                 glucuronidase, beta pseudogene 
                 up 
                 2.01 
                 1.01 
               
               
                   
                 5 
               
               
                 MMRN2 
                 multimerin 2 
                 up 
                 2.00 
                 1.00 
               
               
                 HIF1A 
                 hypoxia inducible factor 1, alpha 
                 up 
                 2.00 
                 1.00 
               
               
                   
                 subunit (basic helix-loop-helix 
               
               
                   
                 transcription factor) 
               
               
                 ZNF271P 
                 zinc finger protein 271, 
                 up 
                 2.00 
                 1.00 
               
               
                   
                 pseudogene 
               
               
                 DBF4B 
                 DBF4 zinc finger B 
                 up 
                 2.00 
                 1.00 
               
               
                 NRCAM 
                 neuronal cell adhesion molecule 
                 up 
                 2.00 
                 1.00 
               
               
                 MKS1 
                 Meckel syndrome, type 1 
                 up 
                 2.00 
                 1.00 
               
               
                 GAREML 
                 GRB2 associated, regulator of 
                 down 
                 −6.04 
                 −2.59 
               
               
                   
                 MAPK1-like 
               
               
                 SNORA2A 
                 small nucleolar RNA, H/ACA 
                 down 
                 −6.04 
                 −2.59 
               
               
                   
                 box 2A 
               
               
                 MUC3A 
                 mucin 3A, cell surface associated 
                 down 
                 −6.04 
                 −2.59 
               
               
                 MAP3K10 
                 mitogen-activated protein kinase 
                 down 
                 −6.12 
                 −2.61 
               
               
                   
                 kinase kinase 10 
               
               
                 SASS6 
                 spindle assembly 6 homolog 
                 down 
                 −6.15 
                 −2.62 
               
               
                   
                 ( C. elegans ) 
               
               
                 BANP 
                 BTG3 associated nuclear protein 
                 down 
                 −6.16 
                 −2.62 
               
               
                 TSPAN15 
                 tetraspanin 15 
                 down 
                 −6.23 
                 −2.64 
               
               
                 SLC25A17 
                 solute carrier family 25 
                 down 
                 −6.27 
                 −2.65 
               
               
                   
                 (mitochondrial carrier; 
               
               
                   
                 peroxisomal membrane protein, 
               
               
                   
                 34 kDa), member 17 
               
               
                 SLC5A1 
                 solute carrier family 5 
                 down 
                 −6.36 
                 −2.67 
               
               
                   
                 (sodium/glucose co transporter), 
               
               
                   
                 member 1 
               
               
                 HYOU1 
                 hypoxia up-regulated 1 
                 down 
                 −6.37 
                 −2.67 
               
               
                 MYH3 
                 myosin, heavy chain 3, skeletal 
                 down 
                 −6.38 
                 −2.67 
               
               
                   
                 muscle, embryonic 
               
               
                 FCRL5 
                 Fc receptor-like 5 
                 down 
                 −6.39 
                 −2.67 
               
               
                 DIS3L2 
                 DIS3 like 3′-5′ exoribonuclease 2 
                 down 
                 −6.43 
                 −2.69 
               
               
                 SUCNR1 
                 succinate receptor 1 
                 down 
                 −6.44 
                 −2.69 
               
               
                 NPBWR1 
                 neuropeptides B/W receptor 1 
                 down 
                 −6.53 
                 −2.71 
               
               
                 MDM2 
                 MDM2 proto-oncogene, E3 
                 down 
                 −6.56 
                 −2.71 
               
               
                   
                 ubiquitin protein ligase 
               
               
                 HIST1H2AC 
                 histone cluster 1, H2ac 
                 down 
                 −6.56 
                 −2.71 
               
               
                 KLRC1 
                 killer cell lectin-like receptor 
                 down 
                 −6.58 
                 −2.72 
               
               
                   
                 subfamily C, member 1 
               
               
                 CDCA7L 
                 cell division cycle associated 7- 
                 down 
                 −6.59 
                 −2.72 
               
               
                   
                 like 
               
               
                 RNA28S5 
                 RNA, 28S ribosomal 5 
                 down 
                 −6.65 
                 −2.73 
               
               
                 MAGT1 
                 magnesium transporter 1 
                 down 
                 −6.68 
                 −2.74 
               
               
                 HYDIN 
                 HYDIN, axonemal central pair 
                 down 
                 −6.68 
                 −2.74 
               
               
                   
                 apparatus protein 
               
               
                 FAM96B 
                 family with sequence similarity 
                 down 
                 −6.70 
                 −2.74 
               
               
                   
                 96, member B 
               
               
                 FPGT 
                 fucose-1-phosphate 
                 down 
                 −6.70 
                 −2.74 
               
               
                   
                 guanylyltransferase 
               
               
                 USP45 
                 ubiquitin specific peptidase 45 
                 down 
                 −6.72 
                 −2.75 
               
               
                 FAM87A 
                 family with sequence similarity 
                 down 
                 −6.72 
                 −2.75 
               
               
                   
                 87, member A 
               
               
                 LMO7DN 
                 LMO7 downstream neighbor 
                 down 
                 −6.78 
                 −2.76 
               
               
                 LACC1 
                 laccase (multicopper 
                 down 
                 −6.80 
                 −2.77 
               
               
                   
                 oxidoreductase) domain 
               
               
                   
                 containing 1 
               
               
                 BAIAP2-AS1 
                 BAIAP2 antisense RNA 1 (head 
                 down 
                 −6.87 
                 −2.78 
               
               
                   
                 to head) 
               
               
                 CACNA1I 
                 calcium channel, voltage- 
                 down 
                 −6.88 
                 −2.78 
               
               
                   
                 dependent, T type, alpha 11 
               
               
                   
                 subunit 
               
               
                 MB21D2 
                 Mab-21 domain containing 2 
                 down 
                 −6.91 
                 −2.79 
               
               
                 AMOTL1 
                 angiomotin like 1 
                 down 
                 −7.01 
                 −2.81 
               
               
                 PALLD 
                 palladin, cytoskeletal associated 
                 down 
                 −7.02 
                 −2.81 
               
               
                   
                 protein 
               
               
                 HTT 
                 huntingtin 
                 down 
                 −7.09 
                 −2.83 
               
               
                 CASC8 
                 cancer susceptibility candidate 8 
                 down 
                 −7.14 
                 −2.84 
               
               
                   
                 (non-protein coding) 
               
               
                 C11orf68 
                 chromosome 11 open reading 
                 down 
                 −7.20 
                 −2.85 
               
               
                   
                 frame 68 
               
               
                 ZNF778 
                 zinc finger protein 778 
                 down 
                 −7.25 
                 −2.86 
               
               
                 FUT7 
                 fucosyltransferase 7 (alpha (1,3) 
                 down 
                 −7.26 
                 −2.86 
               
               
                   
                 fucosyltransferase) 
               
               
                 CT55 
                 cancer/testis antigen 55 
                 down 
                 −7.29 
                 −2.87 
               
               
                 ULK4 
                 unc-51 like kinase 4 
                 down 
                 −7.51 
                 −2.91 
               
               
                 EIF2AK1 
                 eukaryotic translation initiation 
                 down 
                 −7.53 
                 −2.91 
               
               
                   
                 factor 2-alpha kinase 1 
               
               
                 STON1 
                 stonin 1 
                 down 
                 −7.56 
                 −2.92 
               
               
                 NOX1 
                 NADPH oxidase 1 
                 down 
                 −7.63 
                 −2.93 
               
               
                 METAP1D 
                 methionyl aminopeptidase type 
                 down 
                 −7.67 
                 −2.94 
               
               
                   
                 1D (mitochondrial) 
               
               
                 FAM120B 
                 family with sequence similarity 
                 down 
                 −7.68 
                 −2.94 
               
               
                   
                 120B 
               
               
                 IMPG1 
                 interphotoreceptor matrix 
                 down 
                 −7.76 
                 −2.96 
               
               
                   
                 proteoglycan 1 
               
               
                 PBX2 
                 pre-B-cell leukemia homeobox 2 
                 down 
                 −7.77 
                 −2.96 
               
               
                 POLR2F 
                 polymerase (RNA) II (DNA 
                 down 
                 −7.81 
                 −2.97 
               
               
                   
                 directed) polypeptide F 
               
               
                 GLI2 
                 GLI family zinc finger 2 
                 down 
                 −7.93 
                 −2.99 
               
               
                 MRPL28 
                 mitochondrial ribosomal protein 
                 down 
                 −7.94 
                 −2.99 
               
               
                   
                 L28 
               
               
                 STEAP3 
                 STEAP family member 3, 
                 down 
                 −8.08 
                 −3.01 
               
               
                   
                 metalloreductase 
               
               
                 USP2 
                 ubiquitin specific peptidase 2 
                 down 
                 −8.09 
                 −3.02 
               
               
                 SEL1L 
                 sel-1 suppressor of lin-12-like 
                 down 
                 −8.21 
                 −3.04 
               
               
                   
                 ( C. elegans ) 
               
               
                 PCDH17 
                 protocadherin 17 
                 down 
                 −8.25 
                 −3.04 
               
               
                 ASB1 
                 ankyrin repeat and SOCS box 
                 down 
                 −8.30 
                 −3.05 
               
               
                   
                 containing 1 
               
               
                 PER2 
                 period circadian clock 2 
                 down 
                 −8.39 
                 −3.07 
               
               
                 KRT8P12 
                 keratin 8 pseudogene 12 
                 down 
                 −8.45 
                 −3.08 
               
               
                 SYNJ2-IT1 
                 SYNJ2 intronic transcript 1 (non- 
                 down 
                 −8.46 
                 −3.08 
               
               
                   
                 protein coding) 
               
               
                 ADCK5 
                 aarF domain containing kinase 5 
                 down 
                 −8.46 
                 −3.08 
               
               
                 ATP2B1 
                 ATPase, Ca++ transporting, 
                 down 
                 −8.49 
                 −3.09 
               
               
                   
                 plasma membrane 1 
               
               
                 GEMIN2 
                 gem (nuclear organelle) 
                 down 
                 −8.50 
                 −3.09 
               
               
                   
                 associated protein 2 
               
               
                 EIF4B 
                 eukaryotic translation initiation 
                 down 
                 −8.54 
                 −3.09 
               
               
                   
                 factor 4B 
               
               
                 IFNK 
                 interferon, kappa 
                 down 
                 −8.63 
                 −3.11 
               
               
                 SPATS2L 
                 spermatogenesis associated, 
                 down 
                 −8.63 
                 −3.11 
               
               
                   
                 serine-rich 2-like 
               
               
                 SUCLG2-AS1 
                 SUCLG2 antisense RNA 1 (head 
                 down 
                 −8.67 
                 −3.12 
               
               
                   
                 to head) 
               
               
                 OTUD7B 
                 OTU deubiquitinase 7B 
                 down 
                 −8.67 
                 −3.12 
               
               
                 HAPLN2 
                 hyaluronan and proteoglycan link 
                 down 
                 −8.69 
                 −3.12 
               
               
                   
                 protein 2 
               
               
                 TRMT13 
                 tRNA methyltransferase 13 
                 down 
                 −8.72 
                 −3.12 
               
               
                   
                 homolog ( S. cerevisiae ) 
               
               
                 PRR27 
                 proline rich 27 
                 down 
                 −8.73 
                 −3.13 
               
               
                 RCSD1 
                 RCSD domain containing 1 
                 down 
                 −8.81 
                 −3.14 
               
               
                 FBXO24 
                 F-box protein 24 
                 down 
                 −8.99 
                 −3.17 
               
               
                 SYK 
                 spleen tyrosine kinase 
                 down 
                 −9.02 
                 −3.17 
               
               
                 TMEM129 
                 transmembrane protein 129, E3 
                 down 
                 −9.04 
                 −3.18 
               
               
                   
                 ubiquitin protein ligase 
               
               
                 CDRT1 
                 CMT1A duplicated region 
                 down 
                 −9.05 
                 −3.18 
               
               
                   
                 transcript 1 
               
               
                 CDK5RAP3 
                 CDK5 regulatory subunit 
                 down 
                 −9.11 
                 −3.19 
               
               
                   
                 associated protein 3 
               
               
                 OPTN 
                 optineurin 
                 down 
                 −9.13 
                 −3.19 
               
               
                 CYLC1 
                 cylicin, basic protein of sperm 
                 down 
                 −9.21 
                 −3.20 
               
               
                   
                 head cytoskeleton 1 
               
               
                 CAPN3 
                 calpain 3, (p94) 
                 down 
                 −9.35 
                 −3.23 
               
               
                 LARGE-AS1 
                 LARGE antisense RNA 1 
                 down 
                 −9.36 
                 −3.23 
               
               
                 CXorf40B 
                 chromosome X open reading 
                 down 
                 −9.44 
                 −3.24 
               
               
                   
                 frame 40B 
               
               
                 NINL 
                 ninein-like 
                 down 
                 −9.44 
                 −3.24 
               
               
                 ETV5 
                 ets variant 5 
                 down 
                 −9.45 
                 −3.24 
               
               
                 ZNF585A 
                 zinc finger protein 585A 
                 down 
                 −9.59 
                 −3.26 
               
               
                 ARHGAP5-AS1 
                 ARHGAP5 antisense RNA 1 
                 down 
                 −9.63 
                 −3.27 
               
               
                   
                 (head to head) 
               
               
                 ZNF594 
                 zinc finger protein 594 
                 down 
                 −9.65 
                 −3.27 
               
               
                 ALS2 
                 amyotrophic lateral sclerosis 2 
                 down 
                 −9.76 
                 −3.29 
               
               
                   
                 (juvenile) 
               
               
                 SNX10 
                 sorting nexin 10 
                 down 
                 −9.76 
                 −3.29 
               
               
                 LGI2 
                 leucine-rich repeat LGI family, 
                 down 
                 −9.80 
                 −3.29 
               
               
                   
                 member 2 
               
               
                 BTG3 
                 BTG family, member 3 
                 down 
                 −9.93 
                 −3.31 
               
               
                 LCE2A 
                 late cornified envelope 2A 
                 down 
                 −10.02 
                 −3.32 
               
               
                 CC2D1B 
                 coiled-coil and C2 domain 
                 down 
                 −10.29 
                 −3.36 
               
               
                   
                 containing 1B 
               
               
                 DIP2A 
                 DIP2 disco-interacting protein 2 
                 down 
                 −10.43 
                 −3.38 
               
               
                   
                 homolog A ( Drosophila ) 
               
               
                 RFNG 
                 RFNG O-fucosylpeptide 3-beta- 
                 down 
                 −10.55 
                 −3.40 
               
               
                   
                 N-acetylglucosaminyltransferase 
               
               
                 SLN 
                 sarcolipin 
                 down 
                 −10.56 
                 −3.40 
               
               
                 NUP98 
                 nucleoporin 98 kDa 
                 down 
                 −10.65 
                 −3.41 
               
               
                 NSA2 
                 NSA2 ribosome biogenesis 
                 down 
                 −10.70 
                 −3.42 
               
               
                   
                 homolog ( S. cerevisiae ) 
               
               
                 KIF1C 
                 kinesin family member 1C 
                 down 
                 −10.72 
                 −3.42 
               
               
                 ZIC3 
                 Zic family member 3 
                 down 
                 −11.34 
                 −3.50 
               
               
                 MFAP3 
                 microfibrillar-associated protein 
                 down 
                 −11.37 
                 −3.51 
               
               
                   
                 3 
               
               
                 GNA15 
                 guanine nucleotide binding 
                 down 
                 −11.46 
                 −3.52 
               
               
                   
                 protein (G protein), alpha 15 (Gq 
               
               
                   
                 class) 
               
               
                 MAGEA6 
                 melanoma antigen family A, 6 
                 down 
                 −11.60 
                 −3.54 
               
               
                 MCAT 
                 malonyl CoA:ACP 
                 down 
                 −11.65 
                 −3.54 
               
               
                   
                 acyltransferase (mitochondrial) 
               
               
                 MCFD2 
                 multiple coagulation factor 
                 down 
                 −12.07 
                 −3.59 
               
               
                   
                 deficiency 2 
               
               
                 SNORD15A 
                 small nucleolar RNA, C/D box 
                 down 
                 −12.11 
                 −3.60 
               
               
                   
                 15A 
               
               
                 KCNE5 
                 potassium channel, voltage gated 
                 down 
                 −12.25 
                 −3.61 
               
               
                   
                 subfamily E regulatory beta 
               
               
                   
                 subunit 5 
               
               
                 MAGI2-AS3 
                 MAGI2 antisense RNA 3 
                 down 
                 −12.33 
                 −3.62 
               
               
                 NAA60 
                 N(alpha)-acetyltransferase 60, 
                 down 
                 −12.34 
                 −3.63 
               
               
                   
                 NatF catalytic subunit 
               
               
                 GABBR1 
                 gamma-aminobutyric acid 
                 down 
                 −12.73 
                 −3.67 
               
               
                   
                 (GABA) B receptor, 1 
               
               
                 TGFB1I1 
                 transforming growth factor beta 
                 down 
                 −12.84 
                 −3.68 
               
               
                   
                 1 induced transcript 1 
               
               
                 LPAR6 
                 lysophosphatidic acid receptor 6 
                 down 
                 −13.11 
                 −3.71 
               
               
                 RNASE7 
                 ribonuclease, RNase A family, 7 
                 down 
                 −13.47 
                 −3.75 
               
               
                 MAFG 
                 v-maf avian musculoaponeurotic 
                 down 
                 −13.80 
                 −3.79 
               
               
                   
                 fibrosarcoma oncogene homolog 
               
               
                   
                 G 
               
               
                 COL6A4P1 
                 collagen, type VI, alpha 4 
                 down 
                 −15.64 
                 −3.97 
               
               
                   
                 pseudogene 1 
               
               
                 NAV2-AS5 
                 NAV2 antisense RNA 5 
                 down 
                 −15.83 
                 −3.98 
               
               
                 OR52B2 
                 olfactory receptor, family 52, 
                 down 
                 −17.06 
                 −4.09 
               
               
                   
                 subfamily B, member 2 
               
               
                 SLC19A1 
                 solute carrier family 19 (folate 
                 down 
                 −17.20 
                 −4.10 
               
               
                   
                 transporter), member 1 
               
               
                 HIST1H2BM 
                 histone cluster 1, H2bm 
                 down 
                 −17.25 
                 −4.11 
               
               
                 LRRC17 
                 leucine rich repeat containing 17 
                 down 
                 −17.46 
                 −4.13 
               
               
                 TNFRSF10C 
                 tumor necrosis factor receptor 
                 down 
                 −17.69 
                 −4.15 
               
               
                   
                 superfamily, member 10c, decoy 
               
               
                   
                 without an intracellular domain 
               
               
                 ARHGEF7 
                 Rho guanine nucleotide 
                 down 
                 −18.02 
                 −4.17 
               
               
                   
                 exchange factor (GEF) 7 
               
               
                 OR4C6 
                 olfactory receptor, family 4, 
                 down 
                 −18.80 
                 −4.23 
               
               
                   
                 subfamily C, member 6 
               
               
                 CCDC158 
                 coiled-coil domain containing 
                 down 
                 −18.86 
                 −4.24 
               
               
                   
                 158 
               
               
                 RNFT2 
                 ring finger protein, 
                 down 
                 −19.57 
                 −4.29 
               
               
                   
                 transmembrane 2 
               
               
                 MAPKAPK5- 
                 MAPKAPK5 antisense RNA 1 
                 down 
                 −19.79 
                 −4.31 
               
               
                 AS1 
               
               
                 MYO15B 
                 myosin XVB pseudogene 
                 down 
                 −20.06 
                 −4.33 
               
               
                 ANKRD54 
                 ankyrin repeat domain 54 
                 down 
                 −20.15 
                 −4.33 
               
               
                 CLCA4 
                 chloride channel accessory 4 
                 down 
                 −20.19 
                 −4.34 
               
               
                 DNAJB1 
                 DnaJ (Hsp40) homolog, 
                 down 
                 −20.40 
                 −4.35 
               
               
                   
                 subfamily B, member 1 
               
               
                 REEP3 
                 receptor accessory protein 3 
                 down 
                 −21.78 
                 −4.44 
               
               
                 PGBD2 
                 piggyBac transposable element 
                 down 
                 −21.84 
                 −4.45 
               
               
                   
                 derived 2 
               
               
                 CENPV 
                 centromere protein V 
                 down 
                 −22.45 
                 −4.49 
               
               
                 CEP83 
                 centrosomal protein 83 kDa 
                 down 
                 −22.45 
                 −4.49 
               
               
                 TMEM200C 
                 transmembrane protein 200C 
                 down 
                 −24.02 
                 −4.59 
               
               
                 RALBP1 
                 ralA binding protein 1 
                 down 
                 −24.51 
                 −4.62 
               
               
                 DHFRL1 
                 dihydrofolate reductase-like 1 
                 down 
                 −24.69 
                 −4.63 
               
               
                 TDRKH 
                 tudor and KH domain containing 
                 down 
                 −25.59 
                 −4.68 
               
               
                 ALG3 
                 ALG3, alpha-1,3- 
                 down 
                 −26.75 
                 −4.74 
               
               
                   
                 mannosyltransferase 
               
               
                 CKS1B 
                 CDC28 protein kinase regulatory 
                 down 
                 −27.57 
                 −4.78 
               
               
                   
                 subunit 1B 
               
               
                 KIAA1841 
                 KIAA1841 
                 down 
                 −27.85 
                 −4.80 
               
               
                 BHLHE41 
                 basic helix-loop-helix family, 
                 down 
                 −28.68 
                 −4.84 
               
               
                   
                 member e41 
               
               
                 DNAJC3 
                 DnaJ (Hsp40) homolog, 
                 down 
                 −29.22 
                 −4.87 
               
               
                   
                 subfamily C, member 3 
               
               
                 C3orf17 
                 chromosome 3 open reading 
                 down 
                 −29.27 
                 −4.87 
               
               
                   
                 frame 17 
               
               
                 GTF2H5 
                 general transcription factor IIH, 
                 down 
                 −29.36 
                 −4.88 
               
               
                   
                 polypeptide 5 
               
               
                 PTGR1 
                 prostaglandin reductase 1 
                 down 
                 −30.46 
                 −4.93 
               
               
                 JUN 
                 jun proto-oncogene 
                 down 
                 −31.81 
                 −4.99 
               
               
                 TTC28-AS1 
                 TTC28 antisense RNA 1 
                 down 
                 −32.23 
                 −5.01 
               
               
                 CHST2 
                 carbohydrate (N- 
                 down 
                 −32.67 
                 −5.03 
               
               
                   
                 acetylglucosamine-6-O) 
               
               
                   
                 sulfotransferase 2 
               
               
                 KIF22 
                 kinesin family member 22 
                 down 
                 −34.17 
                 −5.09 
               
               
                 CRHR1-IT1 
                 CRHR1 intronic transcript 1 
                 down 
                 −34.51 
                 −5.11 
               
               
                   
                 (non-protein coding) 
               
               
                 ZNF746 
                 zinc finger protein 746 
                 down 
                 −34.76 
                 −5.12 
               
               
                 ZNF658 
                 zinc finger protein 658 
                 down 
                 −36.25 
                 −5.18 
               
               
                 ALS2 
                 amyotrophic lateral sclerosis 2 
                 down 
                 −38.73 
                 −5.28 
               
               
                   
                 (juvenile) 
               
               
                 MTHFD1L 
                 methylenetetrahydrofolate 
                 down 
                 −39.85 
                 −5.32 
               
               
                   
                 dehydrogenase (NADP+ 
               
               
                   
                 dependent) 1-like 
               
               
                 AMACR 
                 alpha-methylacyl-CoA racemase 
                 down 
                 −41.76 
                 −5.38 
               
               
                 COMMD7 
                 COMM domain containing 7 
                 down 
                 −45.82 
                 −5.52 
               
               
                 FIP1L1 
                 factor interacting with PAPOLA 
                 down 
                 −47.06 
                 −5.56 
               
               
                   
                 and CPSF1 
               
               
                 PARP9 
                 poly (ADP-ribose) polymerase 
                 down 
                 −50.35 
                 −5.65 
               
               
                   
                 family, member 9 
               
               
                 HIPK2 
                 homeodomain interacting protein 
                 down 
                 −50.40 
                 −5.66 
               
               
                   
                 kinase 2 
               
               
                 AFF4 
                 AF4/FMR2 family, member 4 
                 down 
                 −50.87 
                 −5.67 
               
               
                 EPB41L4A 
                 erythrocyte membrane protein 
                 down 
                 −52.62 
                 −5.72 
               
               
                   
                 band 4.1 like 4A 
               
               
                 ZNF658 
                 zinc finger protein 658 
                 down 
                 −54.60 
                 −5.77 
               
               
                 EDARADD 
                 EDAR-associated death domain 
                 down 
                 −60.04 
                 −5.91 
               
               
                 PHB 
                 prohibitin 
                 down 
                 −62.21 
                 −5.96 
               
               
                 ZNF532 
                 zinc finger protein 532 
                 down 
                 −67.26 
                 −6.07 
               
               
                 HOXD-AS2 
                 HOXD cluster antisense RNA 2 
                 down 
                 −69.02 
                 −6.11 
               
               
                 DSE 
                 dermatan sulfate epimerase 
                 down 
                 −72.55 
                 −6.18 
               
               
                 NR2C1 
                 nuclear receptor subfamily 2, 
                 down 
                 −75.82 
                 −6.24 
               
               
                   
                 group C, member 1 
               
               
                 ACSF3 
                 acyl-CoA synthetase family 
                 down 
                 −76.12 
                 −6.25 
               
               
                   
                 member 3 
               
               
                 SNORA11C 
                 small nucleolar RNA, H/ACA 
                 down 
                 −76.18 
                 −6.25 
               
               
                   
                 box 11C 
               
               
                 TMEM144 
                 transmembrane protein 144 
                 down 
                 −77.84 
                 −6.28 
               
               
                 CD2AP 
                 CD2-associated protein 
                 down 
                 −83.67 
                 −6.39 
               
               
                 CHML 
                 choroideremia-like (Rab escort 
                 down 
                 −90.20 
                 −6.50 
               
               
                   
                 protein 2) 
               
               
                 RNF2 
                 ring finger protein 2 
                 down 
                 −106.09 
                 −6.73 
               
               
                 HIST1H2BB 
                 histone cluster 1, H2bb 
                 down 
                 −108.05 
                 −6.76 
               
               
                 INTS1 
                 integrator complex subunit 1 
                 down 
                 −112.31 
                 −6.81 
               
               
                 IFT172 
                 intraflagellar transport 172 
                 down 
                 −118.33 
                 −6.89 
               
               
                 PPIA 
                 peptidylprolyl isomerase A 
                 down 
                 −123.27 
                 −6.95 
               
               
                   
                 (cyclophilin A) 
               
               
                 SLC26A6 
                 solute carrier family 26 (anion 
                 down 
                 −125.10 
                 −6.97 
               
               
                   
                 exchanger), member 6 
               
               
                 DOPEY1 
                 dopey family member 1 
                 down 
                 −125.80 
                 −6.98 
               
               
                 SLC22A23 
                 solute carrier family 22, member 
                 down 
                 −131.70 
                 −7.04 
               
               
                   
                 23 
               
               
                 SH3BP4 
                 SH3-domain binding protein 4 
                 down 
                 −132.76 
                 −7.05 
               
               
                 WDR13 
                 WD repeat domain 13 
                 down 
                 −134.84 
                 −7.08 
               
               
                 CSNK1G3 
                 casein kinase 1, gamma 3 
                 down 
                 −141.58 
                 −7.15 
               
               
                 ROM1 
                 retinal outer segment membrane 
                 down 
                 −145.30 
                 −7.18 
               
               
                   
                 protein 1 
               
               
                 MCM4 
                 minichromosome maintenance 
                 down 
                 −148.04 
                 −7.21 
               
               
                   
                 complex component 4 
               
               
                 HHIP-AS1 
                 HHIP antisense RNA 1 
                 down 
                 −158.31 
                 −7.31 
               
               
                 MFAP3L 
                 microfibrillar-associated protein 
                 down 
                 −162.61 
                 −7.35 
               
               
                   
                 3-like 
               
               
                 ALG1 
                 ALG1, 
                 down 
                 −181.56 
                 −7.50 
               
               
                   
                 chitobiosyldiphosphodolichol 
               
               
                   
                 beta-mannosyltransferase 
               
               
                 TAPT1 
                 transmembrane anterior posterior 
                 down 
                 −183.15 
                 −7.52 
               
               
                   
                 transformation 1 
               
               
                 AASDHPPT 
                 aminoadipate-semialdehyde 
                 down 
                 −194.22 
                 −7.60 
               
               
                   
                 dehydrogenase- 
               
               
                   
                 phosphopantetheinyl transferase 
               
               
                 RHOU 
                 ras homolog family member U 
                 down 
                 −207.00 
                 −7.69 
               
               
                 AP1G2 
                 adaptor-related protein complex 
                 down 
                 −208.91 
                 −7.71 
               
               
                   
                 1, gamma 2 subunit 
               
               
                 HSPA4 
                 heat shock 70 kDa protein 4 
                 down 
                 −222.32 
                 −7.80 
               
               
                 HERC2P2 
                 hect domain and RLD 2 
                 down 
                 −257.78 
                 −8.01 
               
               
                   
                 pseudogene 2 
               
               
                 PABPC1 
                 poly(A) binding protein, 
                 down 
                 −285.37 
                 −8.16 
               
               
                   
                 cytoplasmic 1 
               
               
                 ANKRD17 
                 ankyrin repeat domain 17 
                 down 
                 −292.05 
                 −8.19 
               
               
                 SERP1 
                 stress-associated endoplasmic 
                 down 
                 −302.29 
                 −8.24 
               
               
                   
                 reticulum protein 1 
               
               
                 RALGAPB 
                 Ral GTPase activating protein, 
                 down 
                 −322.52 
                 −8.33 
               
               
                   
                 beta subunit (non-catalytic) 
               
               
                 UBE2Q2P1 
                 ubiquitin-conjugating enzyme 
                 down 
                 −326.90 
                 −8.35 
               
               
                   
                 E2Q family member 2 
               
               
                   
                 pseudogene 1 
               
               
                 PLXNA1 
                 plexin A1 
                 down 
                 −423.72 
                 −8.73 
               
               
                 ATXN2 
                 ataxin 2 
                 down 
                 −427.53 
                 −8.74 
               
               
                 ANKRD40 
                 ankyrin repeat domain 40 
                 down 
                 −644.04 
                 −9.33 
               
               
                 RECK 
                 reversion-inducing-cysteine-rich 
                 down 
                 −655.93 
                 −9.36 
               
               
                   
                 protein with kazal motifs 
               
               
                 EIF4E2 
                 eukaryotic translation initiation 
                 down 
                 −682.42 
                 −9.41 
               
               
                   
                 factor 4E family member 2 
               
               
                 CYC1 
                 cytochrome c-1 
                 down 
                 −994.35 
                 −9.96 
               
               
                   
               
            
           
         
       
     
     The above studies, including in vitro experiments and in vivo mouse models of lymphedema, illustrate the potential of therapeutic use of ADSC-EVs for lymphedema treatment. In vitro studies revealed that ADSC-EVs not only have angiogenesis activity, but also lymphangiogeneisis activity. In vivo local injection of ADSC-EVs in lymphedema legs contributed the reduction of enlarged circumference and the induction of capillary vessels and lymphatic vessels. Also, fibrosis of tissue was decreased in ADSC-EVs treatment. Furthermore, the results show that inducing angiogenesis and lymphangiogenesis simultaneously may lead to formation of vessels which expressed both vascular and lymphatic markers and may have function for draining lymphatic system to vascular system, which may work as drainage routes of accumulated fluids and lead to reduction of swelling in lymphedema animal models. Importantly, the results show that lymphedema may be treated with a local application of ADSC-EVs, which also may reduce inflammatory responses, thus decreasing the formation of fibrosis. Results also show that ADSC-EVs contain heterogeneous miRNAs including miR-199a-3p, miR-145-5p, miR-143-3p, miR-377-3p, miR-100-3p, miR-29a-3p, miR-495-3p, and miR-29c-3p which target MDM2, and miR-658, miR-493-3p, miR-184, and miR-27a-3p which target PHB, related to lymphangiogenesis and angiogenesis. MDM2 is a cellular oncoprotein encoded by a gene located on chromosome 12q13-14. MDM2 can suppress p53, a cancer suppressor gene (Duffy, et al.,  Semin Cancer Biol.,  S1044-579X(20):30160-7 (2020)). The inhibition of MDM2 exerts antiinflammatory effects and may lead to the treatment of autoimmune diseases and cancer (Ebrihim, et al.,  Histol Histopathol.,  30(11):1271-82) (2015). Previous studies have shown that MDM2 negatively regulates the stability of hypoxiainducible factor-1 alpha (HIF1a) at the protein level (Shweta, et al.,  J Biol Chem.,  289(33):22785-97 (2014)). The studies reported herein found that the expression level of HIF1a was elevated in LECs after the addition of ADSC-EVs. HIF1a is a transcription factor that controls the cellular response to hypoxia. HIF1a promotes the transcription of various proteins, such as VEGF, Erythropoietin (EPO) and glucose transporters, and plays a key role in lymphangiogenesis and angiogenesis. Thus, these results are consistent with the conclusions that reduced expression of MDM2 caused by eight lymphangiogenic miRNAs in ADSC-EVs led to increase in HIF1a expression in LECs, which caused angiogenesis and lymphangiogenesis. 
     Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of skill in the art to which the disclosed invention belongs. Publications cited herein and the materials for which they are cited are specifically incorporated by reference. 
     Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims.