Reprogramming and Directed Differentiation of Skeletal Muscle Cells from hPSCs.

项目总结/摘要 人类多能干细胞（hPSC）在再生医学方面具有巨大的潜力，因为它们可以 分化成三个胚层的细胞。进一步的hPSC提供了独特的临床前 用于评估患者样品中的疾病机制和疗法的筛选工具。尽管面临巨大 尽管hPSC的潜力很大，但在将干细胞用于基于细胞的治疗之前，需要克服许多障碍。 包括对调节人类骨骼肌的生长因子线索的不完全了解 祖细胞（SMPC）的规格，生长和植入。在这个建议中，我们将研究细胞 在胚胎发育过程中观察到的从hPSC引导骨骼肌命运所需的线索。我们还将 开发第一种筛选方法来识别人类SMPC命运的新调节因子。我们必须 了解如何获得SMPC没有遗传或病毒操纵。作为替代方法，我们有 开发了一种独特的非病毒重编程平台，可实现上级递送和重编程 效率，并将用于指定SMPC没有病毒或遗传操作。我们将比较在 DMD小鼠模型中生长因子定向与重编程SMPC的体内植入潜力。 重编程策略的发展，以指导SMPC的命运和信号调节的鉴定 肌肉祖细胞的特化和维持可以提高我们对人类骨骼肌的基本理解。 以及增强我们从人类产生用于肌肉疾病的可扩展祖细胞的能力 包括DMD。

PROJECT SUMMARY/ABSTRACT Human pluripotent stem cells (hPSCs) have enormous promise for regenerative medicine as they can differentiate into cells from all three embryonic germ layers. Further hPSCs provide a unique pre-clinical screening tool for evaluating disease mechanisms and therapies in patient samples. Despite the enormous potential of hPSC, many obstacles need to be overcome before the use of stem cells in cell-based therapy will be realized, including an incomplete knowledge of the growth factor cues regulating human skeletal muscle progenitor cell (SMPC) specification, growth and engraftment. In this proposal we will investigate the cellular cues required to direct skeletal muscle fate from hPSCs as seen during embryonic development. We will also develop the first screening approach to identify new regulators of human SMPC fate. It is imperative that we understand how to obtain SMPCs without genetic or viral manipulation. As an alternative approach we have developed a unique non viral reprogramming platform, which results in superior delivery and reprogramming efficiency, and will be utilized to specify SMPCs without viral or genetic manipulation. We will compare the in vivo engraftment potential of growth factor directed versus reprogrammed SMPCs in a mouse model of DMD. The development of reprogramming strategies to direct SMPC fate and the identification of signals regulating muscle progenitor cell specification and maintenance could improve our basic understanding of human skeletal myogenesis as well as enhance our ability to generate scalable progenitors from humans for muscle disorders including DMD.