AAV Mini-dystrophin Vectors for DMD Gene Therapy

描述(由申请人提供)： 这项建议的目的是设计、测试和生产一种携带有功能的微肌营养不良蛋白基因的重组腺相关病毒(AAV)载体，该载体将在动物模型中进行广泛的体内治疗效果和长期安全性研究。临床级别的载体将在最后一年生产，用于Duchenne肌营养不良症(DMD)的I期基因治疗安全试验。这项建议是高度整合的，基础研究和临床研究的科学家都参与其中，目标是在五年内产生一个优化的AAV载体，用于DMD患者的I期安全性试验。按照FDA的要求，基因载体的设计、测试和生产以及临床前安全性研究将在匹兹堡大学与其他机构合作进行。俄亥俄州立大学的曾傑瑞·R·门德尔博士领导的团队将进行一项平行研究，这项研究对临床试验的成功同样重要，但通过该提案之外的不同资助机制进行。在该研究中，DMD患者队列将在病理、免疫学和分子水平上进行表征，以供未来招募。为了成功地执行这一具有挑战性的翻译研究项目，我们计划实现五个具体目标。前三个目标将致力于优化微型肌营养不良蛋白基因，选择一个小但强大的肌肉特异性启动子，以及研究基因传递方法。后两个目标将集中在临床前载体毒理学研究和GMP条件下临床级别AAV载体的生产。这笔赠款的成功将导致DMD基因治疗方案的IND申请和RAC提交。

DESCRIPTION (provided by applicant): The goal of this proposal is to design, test, and produce a recombinant adeno-associated viral (AAV) vector carrying a functional mini-dystrophin gene which will be extensively studied in vivo for therapeutic efficacy as well as long-term safety in animal models. The clinical grade vector will be produced in the final year for a Phase I gene therapy safety trial for Duchenne muscular dystrophy (DMD). This proposal is highly integrated, involving scientists in both basic research and clinical studies, with the objective of generating an optimized AAV vector in five years for a Phase I safety trial in DMD patients. The design, test and production of the gene vectors and the pre-clinical safety studies, as required by FDA, will be carried out at the University of Pittsburgh in collaboration with other institutions. A parallel study that is equally important for the success of the clinical trial, but through a different funding mechanism outside this proposal, will be conducted by a team led by Dr. Jerry R. Mendell at the Ohio State University, where a DMD patient cohort will be characterized at the pathological, immunological and molecular levels for future recruitment. To successfully execute this challenging translational research project, we plan to accomplish five Specific Aims. The first three Aims will be devoted to optimization of mini-dystrophin genes, selection of a small but strong muscle-specific promoter, and the investigation of gene delivery methods. The last two Aims will concentrate on preclinical vector toxicology study and clinical grade AAV vector production under the GMP conditions. The success of this grant will lead to the IND filing and RAC submission of the DMD gene therapy protocol.