Molecular and Cellular Therapies for Muscular Dystrophy

Duchenne肌营养不良症是由肌营养不良蛋白编码基因突变引起的，是人类最常见和最具破坏性的遗传病之一。这项应用的目标是加强和改进治疗策略，并为新的创新方法发展基础生物学。Jeff Chamberlain将以他在DMD基因治疗方面的专业知识为基础，开发能够向肌肉细胞或通过造血细胞和间充质干细胞传递微型肌营养不良蛋白和/或成肌调节基因的新型载体。一个相关的目标将是探索将躯体干细胞高效转化为骨骼肌的方法。斯坦·弗罗纳将 研究α-dystrobrevin，一种与dystrophin相关的蛋白在退化过程中的作用。缺乏α-dystrobrevin的小鼠通过一种涉及细胞信号改变的机制发展成肌肉营养不良。将确定α-dystrobrevin的关键功能结构域以及与这些结构域相互作用的蛋白质。针对膜靶向的α-营养不良蛋白，将被测试其缓解MDX肌肉营养不良病理的能力。最后，我们将分析在缺乏α-dystrobrevin的情况下，肌肉细胞本身基因表达的变化。Steve Hausehka将测试调节基因盒的体内组织特异性和长期表达，这些基因盒旨在将不同的治疗性cDNA包装到AAV、慢病毒和腺病毒载体中。然后，这些载体将被用来向mdx4cv小鼠营养不良的肌肉运送微型、微型和全长的dystrophin和各种补偿蛋白。行政核心将提供一般支持，并组织一年两次的西雅图肌肉营养不良会议。老鼠生物学核心将为每个项目提供老鼠模型的功能和病理分析方面的协助。这一综合计划项目的成果将为DMD和其他肌肉退行性疾病带来新的治疗方法。

Duchenne muscular dystrophy, caused by mutations in the gene encoding dystrophin, is one of the most prevalent and devastating human genetic diseases. The goal of this application is to enhance and improve therapeutic strategies and to develop the basic biology for new innovative approaches. Jeff Chamberlain will build on his expertise in DMD gene therapy by developing novel vectors able to deliver mini-dystrophins and/or myogenic regulatory genes either to muscle cells or via hematopoietic and mesenchymal stem cells. A related goal will be exploring methods for the efficient conversion of somatic stem cells into skeletal muscle. Stan Froehner will study the role of alpha-dystrobrevin, a dystrophin-associated protein, in the degeneration process. Mice lacking alpha-dystrobrevin develop muscular dystrophy through a mechanism that involves alteration in cellular signaling. The key functional domains of alpha-dystrobrevin and proteins that interact with these domains will be identified. alpha-dystrobrevin, modified to target to the membrane, will be tested for its ability to alleviate the dystrophic pathology in mdx muscle. Finally, changes in gene expression induced in muscle cells per se by the absence of alpha-dystrobrevin will be analyzed. Steve Hausehka will test the in vivo tissue specificity and long-term expression of regulatory gene cassettes designed to be optimal for packaging different therapeutic cDNAs into AAV, Lentiviral, and Adenoviral vectors. These vectors will then be used to deliver micro-, mini-, and full-length dystrophin and a variety of compensatory proteins to the dystrophic muscles of mdx4cv mice. The Administrative Core will provide general support and organize the biannual Seattle Muscular Dystrophy Conference. The Mouse Biology Core will provide assistance with functional and pathological analyses of mouse models to each project. The results derived from this integrated program project will lead to new therapeutic approaches for DMD and other diseases of muscle degeneration.