Making Beta Cells from Hepatocytes

从肝细胞制备β细胞

• 批准号: 8218113
• 负责人: Jonathan M. Slack
• 金额: $ 5.42万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8218113
• 关键词: Address; Adult; Adverse effects; Allografting; Animals; Autologous; Back; Beta Cell; Biopsy; Cell Differentiation process; Cell Therapy; Cells; Chromatin; Clinic; Clinical; Clinical Trials; DNA; Derivation procedure; Development; Developmental Biology; Diabetes Mellitus; Exocrine pancreas; Experimental Diabetes Mellitus; Gene Expression; Genes; Goals; Graft Rejection; Head; Hepatocyte; Histocompatibility Testing; Human; Immunity; Immunosuppression; In Vitro; Individual; Institutes; Insulin; Insulin-Dependent Diabetes Mellitus; Islets of Langerhans Transplantation; Lead; Liver; Location; Logic; Maturation-Promoting Factor; Methods; Methylation; Minnesota; Organ Donor; Organ Transplantation; Pancreas; Patients; Phenotype; Portal vein structure; Process; Production; Protocols documentation; Research Project Grants; Rest; Rodent; Site; Source; Stem cells; Structure of beta Cell of islet; Technology; Therapeutic; Time; Tissues; Translating; Translational Research; Transplantation; Universities; Work; base; blood glucose regulation; cell type; design; embryonic stem cell; experience; feeding; histone modification; induced pluripotent stem cell; innovation; islet; meetings; overexpression; programs; promoter; public health relevance; success; transcription factor

DESCRIPTION (provided by applicant): New sources of pancreatic beta cells are urgently needed to enable more islet transplants to be carried out for serious cases of type 1 diabetes. A source of autologous beta cells would solve the tissue supply problem and also reduce the need for immunosuppression. This proposal describes a translational research project designed to optimize a protocol for making beta cells, or beta-like cells, from hepatocytes. The recent advent of induced pluripotential cell (iPS) technology has demonstrated the potential for using specific transcription factors to reprogram one cell type to another. The long term goal is to develop a reliable supply of beta cells for clinical transplantation. The objective of this application is to optimize methods for production of beta cells, or cells with a phenotype approaching that of the beta cell, starting from normal hepatocytes. The rationale for this study depends on the close developmental similarity of liver and pancreas, and on previous work showing that overexpression of Pdx1 in liver can provoke expression of beta cell genes. A three step process will be developed based on application of iPS technology, used here not to make embryonic stem cells but to transform one differentiated cell type into another. First, induction of de- differentiation. Second, introduction of specific transcription factors. Third, treatment with agents promoting beta cell differentiation. Preliminary results have already identified the best combination of transcription factors for inducing beta cell-specific gene expression, have shown the critical importance of prior morphological de-differentiation of the hepatocytes, and have identified some maturation-promoting factors. The overall goal will be met through achieving each of the following three specific aims, the results on chromatin changes informing the development of the main protocol: 1: To optimize the methods for producing beta-like cells from primary hepatocytes in vitro. 2: To determine the changes that occur at the chromatin level during hepatocyte de-differentiation. 3: To translate the findings to human hepatocytes. Beta-like cells produced by the optimized protocol will be characterized to establish their phenotype and their capacity to cure experimental diabetes. The approach is innovative because it is the first time that iPS technology and the three step design have been applied to this problem. Success in generating a working protocol will lead directly to studies on larger animals and eventually to clinical trials on human patients. PUBLIC HEALTH RELEVANCE: This project is designed to find a new way to make pancreatic beta cells, by reprogramming of hepatocytes. Islet (beta cell) transplantation is a successful method of cell therapy for treatment of severe type 1 diabetes, but the number of patients who can be treated is severely limited by cell supply. A new source of beta cells would enable many more transplants to be carried out.

描述（申请人提供）：迫切需要新的胰腺β细胞来源，以便能够对严重的1型糖尿病病例进行更多的胰岛移植。自体β细胞来源将解决组织供应问题，并减少对免疫抑制的需求。该提案描述了一个转化研究项目，旨在优化从肝细胞制造β细胞或β样细胞的方案。最近出现的诱导多能细胞（iPS）技术已经证明了使用特定转录因子将一种细胞类型重新编程为另一种细胞类型的潜力。长期目标是开发用于临床移植的可靠的β细胞供应。本申请的目的是优化从正常肝细胞开始生产β细胞或表型接近β细胞的细胞的方法。这项研究的基本原理取决于肝脏和胰腺发育的密切相似性，以及先前的研究表明肝脏中 Pdx1 的过度表达可以激发 β 细胞基因的表达。基于 iPS 技术的应用，将开发一个三步工艺，此处使用的不是制造胚胎干细胞，而是将一种分化细胞类型转化为另一种细胞类型。首先，诱导去分化。其次，引入特定的转录因子。第三，用促进β细胞分化的药物进行治疗。初步结果已经确定了诱导β细胞特异性基因表达的最佳转录因子组合，显示了肝细胞先前形态去分化的至关重要性，并确定了一些成熟促进因子。总体目标将通过实现以下三个具体目标中的每一个来实现，染色质变化的结果为主要方案的开发提供信息： 1：优化在体外从原代肝细胞产生β样细胞的方法。 2：确定肝细胞去分化过程中染色质水平发生的变化。 3：将研究结果转化为人类肝细胞。通过优化方案产生的β样细胞将被表征以确定其表型及其治愈实验性糖尿病的能力。该方法具有创新性，因为这是首次将 iPS 技术和三步设计应用于该问题。成功制定工作方案将直接导致对大型动物的研究，并最终对人类患者进行临床试验。 公共健康相关性：该项目旨在寻找一种通过肝细胞重新编程来制造胰腺β细胞的新方法。胰岛（β细胞）移植是治疗严重1型糖尿病的一种成功的细胞疗法，但可治疗的患者数量受到细胞供应的严重限制。 β细胞的新来源将使更多的移植得以进行。