Targeting Dystroglycanopathies using Pluripotent-derived Myogenic Progenitors

使用多能源性肌源性祖细胞靶向肌营养不良症

基本信息

批准号：
10561375
负责人：
Rita C. R. Perlingeiro
金额：
$ 51.61万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-02-10 至 2027-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10561375
关键词：
Address Affect Age Allogenic Autologous Biochemical Biological Assay Cell Compartmentation Cell Therapy Cell Transplantation Cells Chronic Clinical Trials Complication Development Disease model Engraftment Environment Extracellular Matrix Proteins Fibrosis Future Generations Genes Genetic Diseases Human Human Characteristics Immunologic Deficiency Syndromes Impairment In Vitro Injury Investigation Limb-Girdle Muscular Dystrophies MDC1C Mesoderm Methods Molecular Molecular Profiling Mononuclear Mus Muscle Muscle Fibers Muscle satellite cell Muscular Atrophy Muscular Dystrophies Mutation Myopathy Natural regeneration Patients Play Pluripotent Stem Cells Population Regenerative Medicine Respiratory Diaphragm Role Skeletal Muscle Somatic Cell Technology Therapeutic Therapeutic Effect Transplantation Walker-Warburg syndrome alpha Dystroglycan cell type congenital muscular dystrophy design dystroglycanopathy early onset fukutin related protein gene correction glycosylation human pluripotent stem cell in vivo induced pluripotent stem cell mouse model muscle regeneration mutant post-transplant progenitor programs regeneration potential repaired respiratory response satellite cell single-cell RNA sequencing skeletal stem cell therapy tissue degeneration transcriptome

项目摘要

Summary The biochemical hallmark of FKRP-associated dystroglycanopathies is the hypoglycosylation of α-dystroglycan (α-DG), which leads to disruption in the interaction of α-DG with extracellular matrix proteins, ultimately leading to muscle wasting. Recessive mutations in FKRP are associated with a heterogeneous spectrum of muscle disorders, ranging from severe early-onset to mild late-onset limb-girdle muscular dystrophy (LGMD2I) to several forms of congenital muscular dystrophy (MDC1C), including severe Walker-Warburg Syndrome. Respiratory impairment due to loss of diaphragm function is a prominent complication of both LGMD2I and MDC1C. No approved therapy currently exists for dystroglycanopathies. There has been tremendous excitement for the therapeutic potential of reprogrammed induced pluripotent stem (iPS) cells in treating genetic diseases. The premise of this project is that stem cell-based therapy consisting of human skeletal myogenic progenitors derived from iPSCs will replenish diseased muscle with normal functional muscle fibers as well as muscle stem cells, which have the potential to provide long-term therapeutic effect in dystroglycanopathies. We have developed and extensively validated a method to generate engraftable skeletal myogenic progenitors from pluripotent stem cells through conditional expression of Pax3 or Pax7. This approach results in highly efficient generation of therapeutic myogenic progenitors, which when transplanted into dystrophic mice locally or systemically produce large quantities of functional skeletal muscle tissue that incorporates normally into the host muscle. Importantly, a fraction of transplanted cells remains mononuclear, and displays key features of skeletal muscle stem cells, including satellite cell localization, response to re-injury, and contribution to muscle regeneration in secondary transplantation assays. Therefore, our technology comprises a cell therapy to rebuild functional skeletal muscle, robust to future damage, in hosts with muscular dystrophy. We have recently shown that mouse and human PSC-derived myogenic progenitors contribute to significant myofiber and satellite cell repopulation in the immunodeficient FKRPP448L-NSG mouse model that we generated. Of therapeutic relevance, we have evidence of successful delivery of these myogenic progenitors directly into the diaphragm of FKRP mice. In addition, we have developed a universal gene correction strategy for FKRP, applied this to patient-specific WWS and LGMD2I iPSCs, and demonstrated in vitro and in vivo rescue of functional α-DG glycosylation. In this application, we propose studies that are critical for the development of successful therapeutic approaches for dystroglycanopathies, including understanding 1) the effect of the environment on the engraftment of transplanted cells and 2) the long-term functionality and molecular characteristics of human gene edited WWS and unaffected iPSC-derived myogenic progenitors, important for both autologous and allogeneic future therapeutic applications, respectively.

概括 FKRP相关的多糖果病情的生化标志是α-Dystroglycan的降糖基化（α-DG），导致α-DG与细胞外基质蛋白的相互作用中断，最终领先于浪费肌肉。 FKRP中隐性突变与肌肉的异质谱有关疾病，从严重的早期发作到轻度晚期肢体肌肉营养不良（LGMD2I）到几个先天性肌营养不良症（MDC1C）的形式，包括严重的Walker-Warburg综合征。呼吸道由于diaphragm功能的丧失而导致的损害是LGMD2I和MDC1C的明显并发症。不目前存在批准的治疗疗法。对重编程的诱导多能干（IPS）细胞在治疗遗传疾病中的治疗潜力。这该项目的前提是，基于人类骨骼肌祖细胞组成的基于干细胞的疗法来自IPSC将用正常的功能性肌肉纤维以及肌肉干细胞补充肌肉，这有可能在营养不良的肿瘤病中提供长期治疗作用。我们已经发展了并广泛验证了一种从多能茎生成植入骨骼肌祖祖细胞的方法细胞通过PAX3或PAX7的条件表达。这种方法导致高效的生成治疗性肌源性祖细胞，当将其移植到局部或全身产生的营养不良小鼠中大量的功能性骨骼肌组织通常融入宿主肌肉。重要的是，一小部分移植细胞仍然是单核的，并且显示骨骼肌干细胞的关键特征，包括卫星细胞定位，对重伤的反应以及对次级肌肉再生的贡献移植测定。因此，我们的技术包括一种细胞疗法来重建功能性骨骼肌，在肌肉营养不良的宿主中，对未来的损害有力。我们最近表明了老鼠和人 PSC衍生的肌源性祖细胞有助于大量的肌纤维和卫星细胞在我们生成的免疫缺陷FKRPP448L-NSG小鼠模型。具有治疗意义，我们有证据这些肌源祖细胞成功递送到FKRP小鼠的隔膜中。另外，我们已经为FKRP制定了通用基因校正策略，将其应用于患者特异性WWS和LGMD2I IPSC，并在体外和体内拯救功能性α-DG糖基化。在此应用程序中，我们提案研究对于开发成功的治疗方法至关重要综合策略疗法，包括了解1）环境对植入的影响移植细胞和2）人类基因的长期功能和分子特征编辑了WWS 以及不受影响的IPSC衍生的肌源祖细胞，对自体和同种异体未来都很重要治疗应用。