Translational Gene Therapy for Rhodopsin Autosomal Dominant Retinitis Pigmentosa

描述（由申请人提供）：提出一项多研究者、多中心的研究计划，在动物模型（小鼠和狗）中开发和测试基于基因的视网膜疗法，以便转化为由视紫红质基因（RHO）突变引起的常染色体显性RP患者。RHO突变是人类RP最常见的分子鉴定原因之一，其中100多种突变占RP的约12%。该提案分为4个目标：（目标#1）开发病毒载体、启动子、敲低构建体和替代cDNA，并在两种小鼠模型中比较RHO cDNA增强方法与不依赖等位基因的敲低和替代策略的功效；（目标#2）在大型动物模型（狗）中评估这些策略中哪一种提供了最佳的棒救援，（目标#3）为RHO-ADRP患者的基因治疗临床试验制定结果措施，（目标#4）在临床前安全性研究中评估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)，每个模块都有一组特定的目标，以独特但互补的方式为翻译研究做出贡献。M1 （Vector Development）将提供携带敲低（siRNA，核酶）试剂和耐药（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这两种基因治疗方法。M3（大型动物实验支持）将生产狗，并为这项工作提供基础设施资源)。M4（大型动物I-狗-治疗研究）将在自然发生的RHO-ADRP犬模型中测试这两种方法。M5（人类RHO-ADRP）将识别视网膜区域，可以靶向视网膜治疗的患者。M6（动物载体安全性研究）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RPE65-LCA患者的视网膜基因治疗带入了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. PUBLIC HEALTH RELEVANCE: RELEVANCE (See instructions): Past experience in successfully translating a gene therapy approach for LCA to human patients, and the available resources that this consortium of investigators brings, is a unique opportunity to now target RHO- ADRP. This is one of the most common forms of inherited retinal degeneration for which there is currently no specific treatment available.