Molecular and Cellular Therapies for Muscular Dystrophy

描述（由申请人提供）：肌营养不良症，许多是由编码肌营养不良蛋白复合物的蛋白质的基因突变引起的，是最普遍和最具破坏性的人类疾病之一。目前还没有治愈方法，目前减缓肌肉退化的治疗方法基本上无效。这个应用程序的目标是应用几种肌营养不良症的基本知识来开发治疗方法。在项目1中，Jeffrey Chamberlain将分离替代类型的肌源性干细胞，通过基因转移纠正这些细胞中的原发性遗传病变，并探索将这些细胞移植到同基因的营养不良小鼠中。他将从营养不良的肌肉成纤维细胞中产生肌源性干细胞，并探索它们在体外和体内产生新肌肉组织的能力，并探索从营养不良的肌肉中分离的周细胞的治疗用途。在项目2中，Stephen Tapscott将通过检查肌营养不良犬模型中的肌细胞移植来扩展细胞治疗方法。将通过调节信号通路和细胞外基质成分以及基因操作来探索移植供体细胞的迁移和植入的增强。最后，将比较特定肌肉来源的细胞群在体内重建犬骨骼肌的能力。在项目3中，Stephen Hauschka将修改肌肉特异性调控盒，以在人类肌肉培养物中提供高表达。然后在免疫缺陷小鼠中，在AAV和慢病毒递送至人肌肉异种移植物后，将在体内测试经修饰的盒的治疗性蛋白质的表达。克隆卫星细胞测定和异种移植损伤后人肌纤维再生的分析将确定卫星细胞库是否已稳定转导。在项目4中，Stanley Froehner将研究一种新的补偿基因NPC1，该基因可显著降低mdx小鼠肌肉营养不良表型的严重程度。NPC1表型改善的机制及其对LGMD的适用性将被研究。两个核心设施将为参与实验室提供服务。

DESCRIPTION (provided by applicant): The muscular dystrophies, many caused by mutations in genes encoding proteins of the dystrophin complex, are among the most prevalent and devastating human diseases. No cures exist and current treatments that slow muscle degeneration are largely ineffective. The goal of this application is to apply basic knowledge of several muscular dystrophies to developing therapeutic approaches. In project 1, Jeffrey Chamberlain will isolate alternative types of myogenic stem cells, correct the primary genetic lesion in these cells by gene transfer, and explore the use of such cells for transplantation into syngeneic, dystrophic mice. He will generate myogenic stem cells from dystrophic muscle fibroblasts and explore their ability to generate new muscle tissue in vitro and in vivo and explore the therapeutic use of pericytes isolated from dystrophic muscle. In project 2, Stephen Tapscott will expand the cell therapy approach by examining muscle cell transplantation in the canine model of muscular dystrophy. Enhancement of migration and engraftment of transplanted donor cells will be explored by modulating signaling pathways and extracellular matrix components and genetic manipulations. Finally, specific muscle derived cell populations will be compared for their ability to reconstitute canine skeletal muscle in vivo. In project 3, Stephen Hauschka will modify muscle-specific regulatory cassettes to provide high expression in human muscle cultures. Modified cassettes will then be tested in vivo for expression of therapeutic proteins after AAV and Lentiviral delivery to human muscle xenografts in immunodeficient mice. Clonal satellite cell assays and analysis of human muscle fiber regeneration following xenograft injury will determine whether the satellite cell pool has been stably transduced. In project 4, Stanley Froehner will study a new compensatory gene, NPC1, which markedly reduces the severity of the dystrophic phenotype in mdx mouse muscle. The mechanism of NPC1 phenotype amelioration and its applicability to LGMDs will be studied. Two core facilities will serve the participating laboratories.