Project 1: Translational and pre-clinical studies of muscular dystrophy gene therapy using AAV

项目1：使用AAV进行肌营养不良症基因治疗的转化和临床前研究

• 批准号: 10248345
• 负责人: JEFFREY S CHAMBERLAIN
• 金额: $ 64.3万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-07 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10248345
• 关键词: Address; Animal Model; Area; Canis familiaris; Cardiac; Cardiac Myocytes; Cells; Clinic; Clinical Research; Clinical Trials; Data; Databases; Development; Dystrophin; Facioscapulohumeral Muscular Dystrophy; Funding; Future; Gene Delivery; Gene Expression; Gene Expression Regulation; Gene Targeting; Gene therapy trial; Goals; Heart; Hereditary Disease; Human; Immune Evasion; Inherited; Mediating; Messenger RNA; Methods; Modeling; Mus; Muscle; Muscle Cells; Muscle function; Muscular Dystrophies; Mutation Spectra; Myocardium; Natural History; Nature; Neurology; Pathogenicity; Pathology; Patients; Pattern; Performance; Physical Medicine; RNA Interference; RNAi vector; Radiology Specialty; Rattus; Regulator Genes; Research; Research Project Grants; Ribonucleotide Reductase; Safety; Skeletal Muscle; Striated Muscles; Structure; System; Technology; Testing; Therapeutic; Tissues; Translational Research; Work; adeno-associated viral vector; base; clinical infrastructure; clinical trial readiness; design; gene therapy; heart function; heart imaging; imaging study; improved; improved functioning; in vivo; induced pluripotent stem cell; mRNA Expression; micro-dystrophin; mini-dystrophin; model development; mouse model; novel; patient population; preclinical study; prevent; promoter; protein expression; rapid testing; therapeutic RNA; tool; translational study; trial readiness; vector

This translational research project is designed to improve current AAV vector mediated dystrophin gene therapy methods, to adapt those methods for dominant muscular dystrophies, and to enhance our clinical infrastructure to facilitate participation in future AAV gene therapy trials. Our previous efforts have involved years of collaborative studies that have significantly advanced methods and approaches for gene therapy of DMD. They have also led to approaches to begin harnessing inhibitory RNAi and gene targeting to abrogate gene expression in dominantly inherited muscular dystrophies. We have also developed approaches for systemic gene delivery using AAV vectors, and have made important advances in the study of muscle gene regulation that enable muscle-restricted gene expression and immune evasion following AAV vector delivery. The AAV/micro-dystrophin approaches are far enough along that they are entering human clinical trials to assess safety, and primarily in skeletal muscles, efficacy. The micro-clones, however, are still not fully functional and show reduced activity in cardiac muscles compared with skeletal muscles. The gene therapy approaches for FSHD have been slowed by a lack of good animal models and difficulties in adapting RNAi to AAV. Nonetheless these methods are far enough along that it is reasonable and imperative to enhance their application for optimal gene therapy of many different types of muscular dystrophy. Consequently, our specific aims focus on several of the key limitations of current approaches to gene therapy for DMD and FSHD. For DMD, we focus on enhancing approaches that will functionally target both skeletal and cardiac muscles. We will develop improved gene regulatory cassettes active in all striated muscles, as well as exclusively in skeletal or cardiac muscle. Novel mini-and micro-dystrophins with enhanced function in striated muscles will be designed and screened in multiple test systems. We will also test a promising dual vector strategy, combining dystrophin replacement (structural-based therapy) with enhanced contractile performance via increased ribonucleotide reductase (contractile augmentation therapy) to improve cardiac performance. Our second area of focus is to build upon previous AAV studies for DMD and on mouse model development for FSHD to adapt AAV methods for gene therapy of FSHD. These studies will include analysis of patterns of Dux4 expression in muscle cells. They will also focus on developing and testing AAV vector mediated tissue-specific expression of RNAi hairpins targeting Dux4 mRNA in the context of low level mRNA expression in our AAV-DUX4 mouse model. Finally, we plan to develop patient databases and natural history data focused on cardiac function with colleagues at the MDA clinics in Seattle. We will gather and organize data from ongoing cardiac imaging studies, functional readouts, natural history data and mutational spectra in the DMD patient populations served by the Seattle clinics. These studies will expand the trial readiness goals of Project 2 to include DMD patients.

本翻译研究项目旨在改进现有的AAV载体介导的抗肌萎缩蛋白基因 治疗方法，使这些方法适用于显性肌营养不良症，并提高我们的临床 基础设施，以促进参与未来的AAV基因治疗试验。我们之前的努力 多年的合作研究，大大先进的方法和方法的基因治疗， DMD他们还导致了开始利用抑制性RNAi和基因靶向来消除 显性遗传性肌营养不良症的基因表达。我们还开发了各种方法， 利用AAV载体的系统基因递送，并在肌肉基因的研究中取得了重要进展 在AAV载体递送后，能够实现肌肉限制性基因表达和免疫逃避的调节。 AAV/微肌营养不良蛋白方法已经足够沿着，它们正在进入人类临床试验， 评估安全性，主要是在骨骼肌中的有效性。然而，微型克隆仍然没有完全 与骨骼肌相比，在心肌中显示出降低的活性。基因治疗 由于缺乏良好的动物模型以及难以适应RNA干扰，FSHD的方法进展缓慢 AAV。尽管如此，这些方法已经足够沿着，因此加强它们的 应用于许多不同类型的肌营养不良症的最佳基因治疗。因此，我们的具体 目的集中在目前DMD和FSHD基因治疗方法的几个关键限制。为 DMD，我们专注于增强功能性靶向骨骼肌和心肌的方法。我们 将开发出在所有横纹肌中活跃的改进的基因调控盒，以及专门在骨骼肌中 或心肌。新的小型和微型肌营养不良蛋白与增强功能的横纹肌将 在多个测试系统中设计和筛选。我们还将测试一个有前途的双矢量策略， 肌营养不良蛋白替代（基于结构的治疗），通过增加 核糖核苷酸还原酶（收缩增强疗法），以改善心脏性能。我们的第二领域 重点是建立在以前的DMD AAV研究和FSHD小鼠模型开发的基础上， 用于FSHD的基因治疗的AAV方法。这些研究将包括分析Dux 4表达模式， 肌肉细胞他们还将专注于开发和测试AAV载体介导的组织特异性表达， 在我们的AAV-DUX 4小鼠中低水平mRNA表达的背景下靶向Dux 4 mRNA的RNAi发夹 模型最后，我们计划开发以心脏功能为重点的患者数据库和自然病史数据， 西雅图的MDA诊所的同事们。我们将从正在进行的心脏成像中收集和整理数据， 研究，功能读数，自然史数据和DMD患者人群的突变谱 由西雅图诊所提供。这些研究将扩大项目2的试验准备目标，以纳入DMD患者。