Insulin Producing Cells from Amniotic Stem Cells for Diabetes Therapy

羊膜干细胞产生胰岛素的细胞用于糖尿病治疗

• 批准号: 7676078
• 负责人: ANTHONY ATALA
• 金额: $ 102.51万
• 依托单位: WAKE FOREST UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-30 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7676078
• 关键词: Address; Affect; Allogenic; Amniocentesis; Amniotic Fluid; Autologous; Autologous Transplantation; Beta Cell; C-Peptide; Cell Differentiation process; Cell Line; Cell Lineage; Cell Therapy; Cell Transplantation; Cells; Clinical; Clonal Expansion; Development; Diabetes Mellitus; Disease; Embryo; Face; Germ Layers; Goals; Growth Factor; Health Care Costs; Hormones; Human; Immunodeficient Mouse; Immunosuppression; Immunosuppressive Agents; Implant; In Vitro; Insulin; Insulin-Dependent Diabetes Mellitus; Islet Cell; Islets of Langerhans; Islets of Langerhans Transplantation; Kidney Failure; Laboratories; Medical; Modeling; Mothers; Mus; Neuropathy; Non-Insulin-Dependent Diabetes Mellitus; Organ Donor; Pancreas; Patients; Pharmaceutical Preparations; Phase; Phenotype; Plasmid Cloning Vector; Pluripotent Stem Cells; Population; Production; Program Development; Public Health; Regulator Genes; Research; Retinal Diseases; Source; Stem cells; Streptozocin; Structure of beta Cell of islet; System; Testing; Transplantation; Treatment Protocols; Vascular Diseases; amniotic fluid derived stem cell; base; blood glucose regulation; cell type; clinically relevant; diabetes mellitus therapy; diabetic; falls; glucose metabolism; human embryonic stem cell; improved; in vivo; insulin secretion; islet; meetings; nonhuman primate; novel; offspring; patient population; pre-clinical; pregnant; prevent; progenitor; programs; quantum; research study; stem; stem cell differentiation; transcription factor; vector

DESCRIPTION (provided by applicant): Diabetes mellitus is a growing problem worldwide. In the US it affects over 18 million people and results in annual health care costs exceeding $130 billion. Insulin therapy of Type 1 diabetes, and in advanced cases of Type 2 diabetes, does not prevent serious long-term complications including neuropathy, vascular disease, retinopathy and renal failure. Transplantation of pancreatic islets to restore insulin production offers significant promise. However, the supply of donor pancreata falls far short of meeting the medical need. New sources of insulin producing cells will be required to realize the full potential of cell therapy for diabetes. We propose to generate pancreatic beta lineage cells by in vitro differentiation of stem cells isolated from amniotic fluid. These "AFS cells" are capable of both extensive expansion and differentiation into derivatives of all three embryonic germ layers. Our Preliminary Studies showed that mouse AFS cells can yield insulin producing cells and islet-like cell clusters ("neo-islets"), promoted by expression of the pancreatic transcription factor PDX-1. We now propose to produce neo-islets from human and non-human primate (NHP) AFS cells. To efficiently generate insulin producing cells, we will optimize delivery of a plasmid vector to express PDX-1, and will systematically test growth factors and substrates shown previously to promote pancreatic beta cell differentiation. The resulting neo-islets will be compared with authentic human and NHP pancreatic islets using tests developed for clinical transplantation. To assess their ability to restore control of glucose metabolism and production of insulin and C-peptide, neo-islets will be implanted in immunodeficient mice made diabetic with streptozotocin (STZ). Transplantation in STZ-treated NHP will assess the function of neo-islets in a model physiologically more similar to humans. NHP AFS cell lines will be derived after amniocentesis of pregnant mothers. These stem cells will be used to generate neo-islets for autologous transplantation into the corresponding offspring. The same donor cells will be compared in allogeneic recipients using clinically relevant immunosuppression regimens. Successful development of an abundant source of transplantable insulin producing cells potentially would have a profound impact on the treatment of a major public health problem.

描述（由申请人提供）：糖尿病是一个日益严重的问题，在世界范围内。在美国，它影响超过1800万人，每年的医疗保健费用超过1300亿美元。1型糖尿病和2型糖尿病晚期病例的胰岛素治疗不能预防严重的长期并发症，包括神经病变、血管疾病、视网膜病变和肾衰竭。移植胰岛以恢复胰岛素的产生提供了重大的希望。然而，供体胰腺的供应福尔斯远远不能满足医疗需求。需要新的胰岛素产生细胞来源来实现糖尿病细胞治疗的全部潜力。我们建议通过从羊水中分离的干细胞的体外分化来产生胰腺β谱系细胞。这些“AFS细胞”能够广泛扩增和分化为所有三个胚胎胚层的衍生物。我们的初步研究表明，小鼠AFS细胞可以产生胰岛素产生细胞和胰岛样细胞簇（“新胰岛”），胰腺转录因子PDX-1的表达促进。我们现在提出从人类和非人类灵长类动物（NHP）AFS细胞产生新胰岛。为了有效地产生胰岛素产生细胞，我们将优化质粒载体的递送以表达PDX-1，并将系统地测试先前显示的促进胰腺β细胞分化的生长因子和底物。将使用为临床移植开发的测试将所得新胰岛与真实的人和NHP胰岛进行比较。为了评估它们恢复葡萄糖代谢控制和胰岛素和C肽产生的能力，将新胰岛植入用链脲佐菌素（STZ）制成糖尿病的免疫缺陷小鼠中。在STZ处理的NHP中的移植将在生理上更类似于人类的模型中评估新胰岛的功能。NHP AFS细胞系将在妊娠母亲的子宫内膜穿刺术后获得。这些干细胞将用于产生新胰岛，用于自体移植到相应的后代中。将使用临床相关的免疫抑制方案在同种异体受体中比较相同的供体细胞。成功开发可移植胰岛素产生细胞的丰富来源可能会对重大公共卫生问题的治疗产生深远影响。