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: 7500809
• 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 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7500809
• 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; Condition; 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; Numbers; Organ Donor; Pancreas; Patient currently pregnant; 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; Therapeutic immunosuppression; Transplantation; Treatment Protocols; Vascular Diseases; amniotic fluid derived stem cell; base; blood glucose regulation; cell type; clinically relevant; desire; diabetes mellitus therapy; diabetic; falls; glucose metabolism; human embryonic stem cell; improved; in vivo; insulin secretion; islet; nonhuman primate; novel; pre-clinical; prevent; progenitor; programs; quantum; research study; stem; 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）：糖尿病是世界范围内一个日益严重的问题。在美国，它影响着超过 1800 万人，导致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前景。然而，供体胰腺的供应远远不能满足医疗需求。为了充分发挥糖尿病细胞疗法的潜力，需要新的胰岛素产生细胞来源。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”能够广泛扩增并分化成所有三个胚胎胚层的衍生物。我们的初步研究表明，在胰腺转录因子 PDX-1 表达的促进下，小鼠 AFS 细胞可以产生胰岛素生成细胞和胰岛样细胞簇（“新胰岛”）。我们现在建议从人类和非人类灵长类 (NHP) AFS 细胞中生产新胰岛。为了有效地产生胰岛素细胞，我们将优化表达 PDX-1 的质粒载体的递送，并将系统地测试先前显示的生长因子和底物以促进胰腺 β 细胞分化。将使用为临床移植开发的测试将所得的新胰岛与真实的人类和 NHP 胰岛进行比较。为了评估其恢复葡萄糖代谢控制以及胰岛素和 C 肽生成的能力，新胰岛将被植入链脲佐菌素 (STZ) 导致糖尿病的免疫缺陷小鼠体内。移植到经 STZ 处理的 NHP 中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​​在怀孕母亲进行羊膜穿刺术后获得。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。成功开发出丰富的可移植胰岛素产生细胞来源可能会对重大公共卫生问题的治疗产生深远的影响。