hiPS cells derived skeletal muscle progenitors and their extracellular vesicles for treatment of sarcopenia

项目总结 骨骼肌老化会导致骨质疏松症。据认为，石棺减少的部分原因是运力下降。 干细胞，即卫星细胞，用于修复损伤后的骨骼肌。卫星细胞是主要来源 成体肌肉动态平衡和修复的成肌祖细胞。一种潜在的功能障碍替代方案 卫星细胞是一种诱导的多能干细胞，具有分化为骨骼肌的能力。 肌细胞和血管。在这里，我们已经确定了一种高效的小分子，组蛋白吉维诺斯特(givinostat，givi)。 脱乙酰酶抑制剂(HDACi)可将人IPSC转化为肌祖细胞(MPC) 它们高度增殖并产生大量的细胞外小泡(EV)。我们的“药理学” 使用小分子在有限的时间内生成MPC并且不使用 病毒载体是细胞治疗中非常重要的一步。我们建议IPSC 用GIVI从药理上重新编程为MPC将是再生骨肉形成肌肉的最佳有效方法。 在特定的目的1，假说诱导成肌祖细胞(IMPC)从ipSC与新的小鼠 分子对衰老肌肉的再生是有效和安全的，将进行测试；在特定目标2中，假设 在老化的肌肉微环境中加速iMPC的动员和植入刺激肌肉 将测试再生；在特定目标3中，假设来自Givi诱导的MPC的EV恢复活力 将测试老化的肌肉和增强肌肉的再生。如果iMPC的许多再生特性 可以归功于EV，再生医学将发生范式转变，使内源性自我修复成为可能 在石棉减少症中，通过EV在细胞间转移蛋白质、mRNAs和miRNAs(MiRs)。电动汽车来自工程或 修饰后的干细胞高度富含生物活性分子，包括肌源性MIR 激活对肌肉再生很重要的信号通路。这些研究将涉及多学科 将使用最先进的分子生物学、生化、组织化学、 实验动物的免疫组织化学技术和综合生理学 模型和肌肉功能。这项建议在概念上是创新的，因为它解决了结构和 IMPC及其EV的分子特征及其作为iMPC作用的关键生物信使的研究 治疗骨质疏松症。

PROJECT SUMMARY Aging of skeletal muscle results in sarcopenia. It is believed that sarcopenia is in part due to a decreased capacity of stem cells, namely satellite cells, to repair the skeletal muscle after injury. Satellite cells are the major source of myogenic progenitors for adult muscle homeostasis and repair. A potential alternative for dysfunctional satellite cells is induced pluripotent stem cells (iPSC) which have the capacity to differentiate into skeletal muscle myocytes and blood vessels. Here, we have identified a highly efficient small molecule, givinostat (Givi), a histone deacetylase inhibitor (HDACi) which is capable of transforming human iPSC into myogenic progenitor cells (MPC) that are highly proliferative and generate large numbers of extracellular vesicles (EV). Our “pharmacological reprogramming” approach using small molecules to generate MPC in a limited period of time and without use of viral vectors is a very significant step forward in cell-based therapy. We are proposing that iPSC pharmacologically reprogrammed into MPC with Givi will be optimally effective to regenerate sarcopenic muscle. In specific aim 1, the hypothesis that induced myogenic progenitor cells (iMPC) from iPSC with novel small molecules are effective and safe for regeneration of aged muscle will be tested; In specific Aim 2, the hypothesis that accelerated mobilization and engraftment of iMPC in an aged muscle microenvironment stimulate muscle regeneration will be tested; In specific Aim 3, the hypothesis that EV derived from Givi-induced MPC rejuvenate aged muscle and augment muscle regeneration will be tested. If many of the regenerative properties of iMPC can be credited to EV, there will be a paradigm shift in regenerative medicine to enable endogenous self-repair in sarcopenia by cell to cell transfer of proteins, mRNAs, and miRNAs (miRs) by EV. EV from engineered or modified stem cells are highly enriched with bioactive molecules including myogenic miRs responsible for activation of signaling pathways important in muscle regeneration. These studies will involve multidisciplinary approaches which will employ state of the art molecular biology, biochemical, histochemical, immunohistochemical techniques and integrative physiology involving well established experimental animal model and muscle function. This proposal is conceptually innovative because it addresses the structural and molecular characterization of iMPC and their EV and tests their role as key biological messengers of iMPC action in the treatment of sarcopenia.