Improving dystrophin gene therapy through understanding dystrobrevin function

通过了解肌营养不良蛋白功能改善肌营养不良蛋白基因治疗

• 批准号: 8692012
• 负责人: DeWayne Townsend
• 金额: $ 36.57万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2018-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8692012
• 关键词: Address; Binding; Biochemical; Buffers; C-terminal; Cardiac; Cardiomyopathies; Cause of Death; Cell Therapy; Clinical; Complex; Cytoskeleton; Data; Development; Disease; Duchenne muscular dystrophy; Dystrophin; Dystrophin-Associated Proteins; Effectiveness; Family; Functional disorder; Gene Transduction Agent; Gene Transfer; Genes; Goals; Heart; Heart Diseases; Knockout Mice; Knowledge; Mechanics; Mediating; Membrane; Molecular; Mutation; Myocardium; N-terminal; Patients; Physiological; Play; Psychological reinforcement; Role; Sarcoglycans; Signal Transduction; Site; Skeletal Muscle; Skeletal muscle injury; Therapeutic; Translating; United States; Viral; Viral Vector; Work; adapter protein; base; boys; design; dystrobrevin; effective therapy; exon skipping; gene therapy; improved; in vivo; innovation; insight; mdx mouse; public health relevance; syntrophin

DESCRIPTION (provided by applicant):Heart disease is a leading cause of death in patients with Duchenne muscular dystrophy (DMD). DMD is a uniformly fatal disease characterized by severe skeletal muscle injury and cardiomyopathy, resulting from mutations within the dystrophin gene. The clinical importance of the heart requires that dystrophin's function in the myocardium be a central focus in the development of effective therapies for DMD. Dystrophin connects the internal cytoskeleton to the membrane and functions as a mechanical buffer, protecting the membrane from the forces of contraction. The stability of dystrophin's interaction with the membrane is essential to this mechanical function. Importantly, recent studies have demonstrated that the dystrophin associated protein, dystrobrevin (DB) significantly strengthens the dystrophin-membrane interaction in both cardiac and skeletal muscle. This new finding has significant implications for the design of truncated dystrophin constructs intended for delivery by gene therapy vectors. DB binds to both dystrophin and the membrane bound sarcoglycan complex suggesting a potential mechanism for dystrobrevin's reinforcement of the dystrophin-membrane interaction. DB also plays an important role in maintaining the dystrophin signaling complex, primarily through interactions with the signaling adapter protein syntrophin. DB's important role in modulating the dual mechanical and signaling functions of dystrophin, make a full understanding of DB's interactions essential for the design of mu-dystrophin constructs suitable for genetic therapy. Little is known about the physiological mechanisms by which DB potentiates dystrophin's function in vivo. This limited understanding of factors regulating dystrophin's central functions represents an important gap in biomedical knowledge, inhibiting the ability to design fully functional mu- dystrophin constructs. The long-term goal is to develop a fully functional mu-dystrophin constructs that will form the basis of a viral mediated gene therapy for DMD. The overall objectives of this application are to 1) understand the role of DB in stabilizing the dystrophin signaling complex and reinforcing dystrophin's mechanical interaction with the membrane and 2) translate this knowledge into physiologically relevant improvements in the design of mu-dystrophin constructs. The central hypothesis is that DB is essential to forming the full dystrophin signaling complex and significantly strengthens the dystrophin-membrane interaction and that inclusion of a DB binding domain in mu-dystrophin constructs will result in significant improvements of mu-dystrophin functionality within the heart.

描述（由申请人提供）：心脏病是杜氏肌营养不良症（DMD）患者死亡的主要原因。DMD是一种一致致命的疾病，其特征是严重的骨骼肌损伤和心肌病，由肌营养不良蛋白基因内的突变引起。心脏的临床重要性要求肌营养不良蛋白在心肌中的功能成为DMD有效疗法开发的中心焦点。肌营养不良蛋白将内部细胞骨架连接到膜上，并作为机械缓冲剂，保护膜免受收缩力的影响。抗肌萎缩蛋白与膜相互作用的稳定性对于这种机械功能是必不可少的。重要的是，最近的研究表明，肌营养不良蛋白相关蛋白，dystrobrevin（DB）显着加强心肌和骨骼肌中的肌营养不良蛋白-膜相互作用。这一新发现对设计截短的抗肌萎缩蛋白构建体具有重要意义，该构建体旨在通过 基因治疗载体DB结合到肌营养不良蛋白和膜结合的肌聚糖复合物，这表明肌营养不良蛋白-膜相互作用的肌营养不良蛋白的强化的潜在机制。DB还在维持肌营养不良蛋白信号传导复合物中起重要作用，主要通过与信号传导衔接蛋白突触营养蛋白的相互作用。DB在调节抗肌萎缩蛋白的双重机械和信号传导功能中的重要作用，使得充分理解DB的相互作用对于设计适合于遗传治疗的μ-抗肌萎缩蛋白构建体至关重要。关于DB增强抗肌萎缩蛋白在体内的功能的生理机制知之甚少。对调节肌营养不良蛋白的中心功能的因子的这种有限的理解代表了生物医学知识中的重要空白，抑制了设计全功能的抗肌营养不良蛋白构建体的能力。长期目标是开发一种全功能的mu-肌营养不良蛋白构建体，其将形成DMD病毒介导的基因治疗的基础。本申请的总体目标是1）理解DB在稳定抗肌萎缩蛋白信号传导复合物和增强抗肌萎缩蛋白与膜的机械相互作用中的作用，和2）将该知识转化为μ-抗肌萎缩蛋白构建体的设计中的生理学相关的改进。中心假设是DB对于形成完整的肌营养不良蛋白信号传导复合物是必不可少的，并且显著增强肌营养不良蛋白-膜相互作用，并且在μ-肌营养不良蛋白构建体中包含DB结合结构域将导致心脏内μ-肌营养不良蛋白功能的显著改善。