Large animal therapy studies

提出了一项多研究人员、多中心的研究计划，以开发和测试基于基因的视网膜疗法 突变引起的常染色体显性遗传性RP患者的动物模型(小鼠和狗) 在视紫红质基因(Rho)中。Rho突变构成了最常见的分子鉴定之一 人的致病因素，超过100种，占致病因素的12%。这项提议存在分歧。 4个目标将：(目标1)开发病毒载体、启动子、击倒结构和替代 CDNA，并比较Rho cDNA扩增方法和非等位基因非依赖方法的有效性 在两个小鼠模型中的击倒和替换策略；(目标2)在大型动物模型(狗)中进行评估 这些策略中哪一种提供了抢救RODS的最佳方案？(目标3)为临床制定结果衡量标准 Rho-ADRP患者的基因治疗试验，以及(目标4)评估最优策略和载体 在临床前安全性研究中构建(基于AIMS#1和2的结果)。六个协调模块(M)是 描述，每个都有一组特定的目标，这些目标以独特但互补的方式对 翻译研究。M1(载体开发)将提供携带敲除基因(siRNA、核酶)的AAVs 试剂，以及抗性(硬化)的Rho cDNA。M2(小动物-老鼠疗法研究)将测试2 两种小鼠模型的基因治疗方法。M3(大型动物实验支持)将产生 并为这项工作提供基础设施资源)。M4(大型动物i-狗疗法研究)将测试 两种方法在自然发生的犬Rho-ADRP模型中。M5(人类Rho-adrp)将 确定可作为患者视网膜局部治疗目标的视网膜区域。M6(传病媒介安全研究 动物)将在小动物和大动物中进行基于GLP的临床前毒理学和生物分布研究 测试最佳(“铅”)治疗载体的安全性，作为FDA考虑的基本第一步 为未来的I期临床试验提供IND。本提案中描述的研究研究代表了 延续了模块科学家之间的长期合作，已经带来了视网膜 RPE65-LCA患者的基因治疗进入I期临床试验。

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 fro 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.