Molecular and Cellular Therapies for Muscular Dystrophy

由编码肌营养不良蛋白的基因突变引起的杜氏肌营养不良症是最普遍和最具破坏性的人类遗传性疾病之一。该应用程序的目标是增强和改进治疗策略，并为新的创新方法开发基础生物学。Jeff Chamberlain将利用他在DMD基因治疗方面的专业知识，开发能够将迷你肌营养不良蛋白和/或肌源性调节基因递送到肌肉细胞或通过造血和间充质干细胞的新型载体。一个相关的目标将是探索将体干细胞有效转化为骨骼肌的方法。Stan Froehner将 研究α-肌营养不良蛋白，一种肌营养不良蛋白相关蛋白，在变性过程中的作用。缺乏α-短肌营养不良蛋白的小鼠通过涉及细胞信号传导改变的机制发展肌营养不良症。将确定α-肌营养不良蛋白的关键功能结构域和与这些结构域相互作用的蛋白质。将测试经修饰以靶向膜的α-短肌营养不良蛋白减轻mdx肌肉中营养不良病理的能力。最后，将分析由于α-肌养短蛋白的缺乏而在肌细胞本身中诱导的基因表达的变化。Steve Hausehka将测试调节基因盒的体内组织特异性和长期表达，该基因盒设计用于将不同的治疗性cDNA包装到AAV、慢病毒和腺病毒载体中。然后，这些载体将用于将微小、迷你和全长肌营养不良蛋白和各种补偿蛋白递送到mdx 4cv小鼠的营养不良肌肉。 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。小鼠生物学核心将为每个项目提供小鼠模型的功能和病理分析。这项综合项目的结果将为DMD和其他肌肉变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.