Derivation of pancreatic beta cells from human induced pluripotent stem cells

从人类诱导多能干细胞衍生胰腺β细胞

• 批准号: 8625785
• 负责人: Weidong Wang
• 金额: $ 34.44万
• 依托单位: OKLAHOMA MEDICAL RESEARCH FOUNDATION
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8625785
• 关键词: Address; Agonist; Autologous; B cell differentiation; B-Cell Development; B-Lymphocytes; Biological; Blood Glucose; C-Peptide; Cell Culture Techniques; Cell Lineage; Cell Therapy; Cell Transplantation; Cell surface; Cells; Code; Complex; Cultured Cells; Cytoplasmic Granules; Derivation procedure; Development; Developmental Biology; Diabetes Mellitus; Diabetic mouse; DsRed; Engineering; Event; Gene Expression; Gene Expression Profile; Gene Targeting; Genes; Genetic; Glucose; Goals; Human; In Vitro; Instruction; Insulin; Insulin-Dependent Diabetes Mellitus; Islets of Langerhans; Islets of Langerhans Transplantation; Knock-in Mouse; Knowledge; Methods; Modeling; Molecular; Molecular Profiling; Mus; Oklahoma; Pancreas; Pathway interactions; Pluripotent Stem Cells; Population; Protocols documentation; Reading; Replacement Therapy; Reporter; Research; Reverse Transcriptase Polymerase Chain Reaction; Role; Signal Pathway; Signal Transduction; Site; Sorting - Cell Movement; Source; Staging; Stem cells; Structure of beta Cell of islet; Technology; Testing; Time; Transcription Coactivator; Transplantation; cell type; expectation; experience; human embryonic stem cell; improved; in vivo; induced pluripotent stem cell; insight; interest; islet; mouse model; notch protein; novel; nuclease; pancreas development; progenitor; programs; promoter; research study; restoration

PROJECT SUMMARY (See instructions): Pancreatic islet transplantation provides a promising therapy for Type 1 Diabetes mellitus (Tl DM), in which the majority of pancreatic insulin-producing p-cells are destroyed. The shortage of transplantable donor islets has stimulated much interest in using human pluripotent stem cells such as human embryonic stem cells and human induced pluripotent stem cells (hiPSCs) as alternative, renewable sources to generate functional B-cells. Our long-term goal is to develop an autologous cell-based therapy to replenish insulin producing B-cells for the purpose of treating T1 DM by inducing the differentiation of hiPSCs. The remarkable developmental and differentiation potential of hiPSCs makes them attractive candidates for cell-based therapies. Before the full potential of hiPSCs can be realized, however, it is necessary to understand the complex signaling and genetic mechanisms that control their differentiation. This proposal will employ hiPSC as an exciting new model to dissect the molecular and cellular programs that regulate human B-cells differentiation. Through use of a novel TALEN technology to make knock-in reporter hiPSC lines, the proposed research will develop methods for isolating human B-cells progenitors and tracing the fates of their differentiated progeny. The proposed studies are aimed at defining and characterizing late stage progenitors of B-cell development, establishing their lineage relationships, and identifying cell type-specific cell surface markers of these progenitor populations and signaling pathways that guide their fate. These will be addressed by pursuing three Specific Aims: 1) Develop cell type-specific reporter hiPSCs for ordering events in human B-cells differentiation. 2) Determine the roles of Wnt, Notch and other critical signaling pathways in human B-cells differentiation. 3) Establish the lineage relationships and differentiation potentials of hiPSC derived B-cell sprogenitor subsets. Using genetic, cell biological and in vivo approaches, the planned research will improve our understanding of the molecular and cellular programs underlying human B-cells differentiation, maturation and function.

项目摘要（参见说明）： 胰岛移植为 1 型糖尿病 (T1 DM) 提供了一种有希望的治疗方法，其中大多数产生胰岛素的胰腺 p 细胞被破坏。可移植供体胰岛的短缺激发了人们对使用人类多能干细胞（例如人类胚胎干细胞和人类诱导多能干细胞（hiPSC））作为替代的可再生来源来产生功能性 B 细胞的浓厚兴趣。我们的长期目标是开发一种基于自体细胞的疗法来补充产生胰岛素的 B 细胞，从而通过诱导 hiPSC 分化来治疗 T1 DM。 hiPSC 卓越的发育和分化潜力使其成为基于细胞的疗法的有吸引力的候选者。然而，在充分发挥 hiPSC 的潜力之前，有必要了解控制其分化的复杂信号传导和遗传机制。该提案将采用 hiPSC 作为令人兴奋的新模型来剖析调节人类 B 细胞分化的分子和细胞程序。通过使用新型 TALEN 技术来制造敲入报告基因 hiPSC 系，拟议的研究将开发分离人类 B 细胞祖细胞并追踪其分化后代命运的方法。拟议的研究旨在定义和表征 B 细胞发育的晚期祖细胞，建立它们的谱系关系，并鉴定这些祖细胞群的细胞类型特异性细胞表面标记和指导其命运的信号通路。这些问题将通过追求三个具体目标来解决：1) 开发细胞类型特异性报告基因 hiPSC，用于对人类 B 细胞分化中的事件进行排序。 2) 确定Wnt、Notch和其他关键信号通路在人类B细胞分化中的作用。 3) 建立 hiPSC 衍生的 B 细胞祖细胞亚群的谱系关系和分化潜力。计划中的研究将利用遗传、细胞生物学和体内方法，增进我们对人类 B 细胞分化、成熟和功能背后的分子和细胞程序的理解。