Translational Gene Therapy for Rhodopsin Autosomal Dominant Retinitis Pigmentosa

描述（由申请方提供）：拟定了一项多研究者、多中心研究计划，以在动物模型（小鼠和犬）中开发和测试基于基因的视网膜治疗，用于治疗由视紫红质基因（RHO）突变引起的常染色体显性遗传RP患者。RHO突变构成了人类RP最常见的分子鉴定原因之一，其中超过100种占RP的> 12%。（目标#1）开发病毒载体、启动子、敲低构建体和置换cDNA，并在两种小鼠模型中比较RHO cDNA扩增方法与等位基因非依赖性敲低和置换策略的功效;（目标#2）在大型动物模型（狗）中评估这些策略中的哪一种提供了对视杆细胞的最佳拯救，（目标#3）开发用于RHO-ADRP患者中基因治疗的临床试验的结果测量，和（目标4）评价临床前安全性研究中的最佳策略和载体构建（基于目标1和2的结果）。描述了六个协调模块（M），每个模块都有一套特定的目标，以独特但互补的方式对翻译研究做出贡献。M1（载体开发）将提供携带敲低（siRNA、核酶）试剂和抗性（硬化）RHO cDNA的AAV。M2（小动物-小鼠-治疗研究）将在两种小鼠模型中测试2种基因治疗方法。M3（大型动物实验支持）将生产犬，并为这项工作提供基础设施资源。M4（大型动物I-犬-治疗研究）将在天然存在的RHO-ADRP犬模型中测试2种方法。M5（人RHO-ADRP）将识别可用于患者局部视网膜治疗的视网膜区域。M6（动物中的载体安全性研究）将在小型和大型动物中进行基于GLP的临床前毒理学和生物分布研究，以测试最佳（“先导”）治疗载体的安全性，作为FDA考虑IND用于未来I期临床试验的重要第一步。本提案中描述的研究代表了模块科学家之间长期合作的延续，该合作已经将RPE 65-LCA患者的视网膜基因治疗带入I期临床试验。

DESCRIPTION (provided by applicant): A multi-investigator, multi-center research plan is proposed to develop and test gene-based retinal therapy in animal models (mouse and dog) for translation to patients with autosomal dominant RP caused by mutations in the rhodopsin gene (RHO). RHO mutations constitute one of the most common molecularly-identified causes of human RP, and more than 100 of them account for > 12 % of RP. The proposal has been divided into 4 aims that will: (Aim#1) develop viral vectors, promoters, knockdown constructs and replacement cDNAs, and compare the efficacy of a RHO cDNA augmentation approach, to that of an allele-independent knockdown and replacement strategy in two mouse models; (Aim #2) evaluate in a large animal model (dog) which of these strategies provides optimal rescue of rods, (Aim #3) develop outcome measures for clinical trials of gene therapy in RHO-ADRP patients, and (Aim #4) evaluate the optimal strategy and vector construct (based on results of Aims #1 and 2) in pre-clinical safety studies. Six coordinated modules (M) are described, each with a specific set of aims that contributes in a unique but complementary way to the translational studies. M1 (Vector Development) will provide AAVs carrying knockdown (siRNA, ribozymes) reagents, and resistant (hardened) RHO cDNAs. M2 (Small Animal-mouse- Therapy Studies) will test the 2 gene therapy approaches in two mouse models. M3 (Large Animal Experiemntal Support) will produce the dogs, and provide infrastructure resources for this work). M4 (Large anima I- dog - Therapy Studies) will test the 2 approaches in a naturally -occurring canine model of RHO-ADRP. M5 (Human RHO-ADRP) will identify retinal regions that can be targeted for focal retinal therapy in patients. M6 (Vector safety studies in Animals) will conduct GLP-based preclinical toxicology and biodistribution studies in small and large animals to test the safety of the optimal ("lead") therapeutic vector as the essential first step ro FDA consideration of an IND for a future Phase I Clinical Trial. The research studies described in this proposal represent a continuation of a longstanding collaboration between the module scientists that already has brought retinal gene therapy for RPE65-LCA patients to a Phase I clinical trial.