Large animal therapy studies

提出了一项多研究者、多中心的研究计划，以开发和测试基于基因的视网膜治疗， 动物模型（小鼠和犬），用于转化为突变引起的常染色体显性遗传RP患者 视紫红质基因（Rhodopsin Gene，RHO）RHO突变构成了最常见的分子鉴定 引起人类RP的原因有100多种，占RP的12%以上。该提案已被分为 分为4个目标，将：（目标#1）开发病毒载体，启动子，敲除构建体和替换 cDNA，并比较RHO cDNA扩增方法与等位基因非依赖性扩增方法的功效。 在两种小鼠模型中敲除和替换策略;（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棒的最佳挽救，（目标#3）制定临床结局指标 在RHO-ADRP患者中进行基因治疗的试验，并（目的#4）评估最佳策略和载体 临床前安全性研究中的结构（基于目标1和2的结果）。六个协调模块（M）是 每一个都有一套特定的目标，以一种独特但互补的方式促进 翻译研究M1（载体开发）将提供携带敲减（siRNA、核酶）的AAV 试剂和抗性（硬化）RHO cDNA。M2（小动物-小鼠-治疗研究）将测试2种 在两个小鼠模型中的基因治疗方法。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-犬-治疗研究）将进行试验 在自然发生的RHO-ADRP犬模型中的2种方法。M5（人RHO-ADRP）将 识别可以作为患者中的局部视网膜治疗的目标的视网膜区域。M6（在非洲开展的媒介安全性研究） 动物）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 测试最佳（“先导”）治疗载体的安全性，作为FDA考虑的重要第一步， 用于未来I期临床试验的IND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， RPE 65-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.