Innovative Approaches to Treat Duchenne Muscular Dystrophy Using iPSC-Derived Muscle Progenitors

 描述(申请人提供)：Duchenne肌营养不良症(DMD)是一种致命的肌肉萎缩疾病，由于缺乏肌营养不良蛋白，最终导致肌肉细胞凋亡和肌肉收缩能力受损。诱导多能干细胞(IPSC)提供了纠正DMD基因缺陷和创造健康细胞进行移植的可能性，而不会引起免疫激活。然而，这需要工程IPSCs在不促进肿瘤形成的情况下促进归巢和植入到患病的肌肉组织中。我们的长期目标是优化IPSC向肌源性祖细胞(MPC)的分化，这些MPC被定向定向到肌肉组织中，并在不受限制的生长的情况下植入。我们的中心假设是，精确的MPC基因工程，再加上通过预适应或外周小体输送对宿主肌肉利基进行靶向修饰，将促进供体IPSC来源的MPC通过SDF-CXCR4轴的归巢和植入，从而安全有效地治疗DMD。其基本原理是开发方法学，从DMD患者那里生产大量功能性MPC，这些MPC是无肿瘤的，可以有效地定位肌肉损伤部位并促进修复。我们计划测试我们的中心假设，并通过追求以下三个具体目标来实现此应用程序的目标。在目标1中，我们将验证IPSC来源的祖细胞对于营养不良肌肉的再生是有效和安全的假设。在目标2中，我们将验证这样一种假设，即使用基于CRISPR的基因组平台激活DMD小鼠iPSC来源的肌源性前体细胞中的CXCR4可以促进供体细胞在DMD小鼠体内的植入。在目标3中，我们将验证这样一种假设，即通过缺血预适应或ipsc-exosome介导的SDF-1α蛋白传递来优化肌肉再生微环境，提高供体MPC的归巢和/或存活，以促进肌肉修复。这些实验有可能证明，移植IPSC来源的祖细胞，结合优化宿主肌肉微环境的方法，将更有效地改善营养不良病理，提高DMD患者的生活质量。

 DESCRIPTION (provided by applicant): Duchenne Muscular Dystrophy (DMD) is a lethal muscle wasting disease caused by the lack of dystrophin, which eventually leads to apoptosis of muscle cells and impaired muscle contractility. Induced pluripotent stem cells (iPSC) offer the potential to correct the DMD gene defect and create healthy cells for transplantation without causing immune activation. However, this requires engineering iPSCs to facilitate homing and engraftment into diseased muscle tissue without promoting tumorigenesis. Our long-term goal is to optimize the differentiation of iPSC into myogenic progenitor cells (MPC) that are directed to home into muscle tissue and engraft without undergoing unrestrained growth. Our central hypothesis is that precise MPC genetic engineering, coupled with targeted modification of the host muscle niche by preconditioning or exosome delivery, will boost homing and engraftment of donor iPSC-derived MPC via the SDF-CXCR4 axis to safely and effectively treat DMD. The rationale is to develop methodology to produce large quantities of functional MPC from patients with DMD that are tumor free and can effectively home to sites of muscle injury and facilitate repair. We plan to test our central hypothesis and accomplish the objectives of this application by pursuing the following three specific aims. In Aim 1, we will test the hypothesis that iPSC-derived progenitor cells are effective and safe for regeneration of dystrophic muscle. In Aim 2, we will test the hypothesis that activation of CXCR4 in iPSC-derived myogenic precursors from DMD mice using a CRISPR-on based genomic platform improves engraftment of donor cells in DMD mice. In Aim 3, we will test the hypothesis that optimizing the regenerative microenvironment in muscle by ischemic preconditioning or iPSC-exosome-mediated delivery of SDF-1α protein enhances the homing and/or survival of donor MPC to augment muscle repair. These experiments have the potential to demonstrate that transplantation of iPSC-derived progenitors, coupled with methods to optimize the host muscle microenvironment, will more effectively ameliorate dystrophic pathology and improve the quality of life for patients with DMD.