Derivation of pancreatic beta cells from human induced pluripotent stem cells

项目总结(见说明)： 胰岛移植为1型糖尿病提供了一种很有前途的治疗方法，在1型糖尿病中，大部分产生胰岛素的胰岛β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如人类胚胎干细胞和人类诱导多能干细胞(HiPSCs)作为替代的、可再生的来源来产生功能性B细胞的兴趣。我们的长期目标是开发一种基于细胞的自体疗法，以补充产生胰岛素的B细胞，以通过诱导hPSCs分化来治疗T1 DM。HPSCs显着的发育和分化潜力使其成为基于细胞治疗的有吸引力的候选细胞。然而，在充分发挥HiPSCs的潜力之前，有必要了解控制其分化的复杂信号和遗传机制。这项提议将利用HiPSC作为一个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。通过使用一种新的TALEN技术来制造敲入报告hPSC系，这项拟议的研究将开发分离人类B细胞前体细胞并追踪其分化后代的命运的方法。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，并确定这些祖细胞群体的特定细胞类型的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)开发细胞类型特异的报告分子HiPSCs，用于对人类B细胞分化过程中的事件进行排序。2)确定Wnt、Notch等关键信号通路在人类B细胞分化中的作用。3)建立HiPSC来源的B细胞祖细胞亚群的谱系关系和分化潜能。利用遗传学、细胞生物学和体内方法，计划中的研究将提高我们对人类B细胞分化、成熟和功能背后的分子和细胞程序的理解。

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.