Reprogramming and Directed Differentiation of Skeletal Muscle Cells from hPSCs

摘要 人类多能干细胞(HPSCs)为理解人类肌肉发生和发展提供了巨大潜力 在盘子里模拟疾病。在上一个资助期，我们已经确定了 来源于hPSC的骨骼肌祖细胞(SMPC)在胚胎至胎儿7-12周左右 在人类发展中。我们发现SmPC不能有效地移植到小鼠的壁龛中，而成体 肌肉干细胞(SCs)具有这种强大的潜力。随着MDX小鼠模型中疾病严重性的增加 在Duchenne肌营养不良症中，我们发现微环境中的异常细胞增加，改变强劲 嫁接。这项提案中的下一阶段工作现在集中在提高我们的调节能力 SmPC和SC状态，并改善植入到小鼠的利基，作为真正的再生干细胞。通过使用 作为人类发展的典范，我们的工作已经为我们理解 人类肌肉发生和生肌细胞的产生。我们将利用这些工具、测序数据集和 在上一个供资期间获得的知识，以调整SMPC和SC状态并产生更好的 这次更新中的治疗候选者。我们将定义并从功能上验证关键监管网络 对控制胚胎、胎儿Smpc或成年干细胞很重要，并提高我们对如何 祖细胞和干细胞在人类肌肉发生和hPSCs发生过程中随时间变化(目标1)。更进一步，我们 将评估从hPSC SMPC调节生态位占用的重要参数(目标2)。这部作品 将提高我们产生健壮干细胞的能力，以改进疾病建模和用于再生 医药。

ABSTRACT Human pluripotent stem cells (hPSCs) offer enormous potential for understanding human myogenesis and modeling diseases in a dish. In the previous funding period, we have identified the developmental trajectory of skeletal muscle progenitor cells (SMPCs) derived from hPSCs to be around embryonic to fetal week 7-12 stage in human development. We showed that SMPCs do not engraft into the mouse niche efficiently whereas adult muscle stem cells (SCs) have this robust potential. Further as disease severity increases in mdx mouse models of Duchenne Muscular Dystrophy we found an increase in aberrant cells in the microenvironment altering robust engraftment. The next phase of this work in this proposal is now focused on improving our ability to modulate SMPC and SC states and improve engraftment into the mouse niche as true repopulating stem cells. By using human development as a model, our work has already yielded significant insights into our understanding of human myogenesis and the generation of myogenic cells. We will utilize the tools, sequencing datasets and knowledge gained in the previous funding period to modulate SMPC and SC states and generate a better therapeutic candidate in this renewal. We will define and functionally validate the key regulatory networks important for controlling embryonic, fetal SMPCs, or adult SCs and improve our basic knowledge of how progenitors and stem cells change over time during human myogenesis and from hPSCs (Aim 1). Further we will evaluate the parameters important for modulating niche occupancy from hPSC SMPCs (Aim 2). This work will improve our ability to generate a robust stem cell for improved disease modeling and use in regenerative medicine.