AUTOSOMAL DOMINANT EYE DISEASE: CATALYTIC RNA TREATMENT

描述（由申请人提供）：本项目的目标是开发一个 常染色体显性视网膜色素变性（ADRP）的遗传治疗。视网膜炎 色素沉着症是一种遗传性视网膜变性， 视杆细胞每3500人中约有1人受影响， 视力逐渐丧失，最终失明，通常在一段时间内， 几十年超过30个基因的突变导致视网膜色素变性，但缺陷 在视紫红质基因中，视杆细胞的主要捕光色素， 是ADRP的主要原因。这些显性突变导致 故障，错误分类或折叠不良的分子，最终杀死 我们的基本假设是， 这些突变形式的视紫红质的表达可以拯救视杆细胞光感受器 保护视力我们将为此目的使用的遗传工具是 催化RNA分子或核酶。小的核酶可以被改造成切断 几乎所有的RNA都能以序列特异性的方式表达。我们用来 将核酶引入视网膜细胞是重组腺相关病毒， 病毒或AAV。在目前的资助期间，我们设计了核酶， 特异于ADRP转基因大鼠模型中存在的突变型视紫红质mRNA。 当这些锤头鲨被运送到携带P23 H视紫红质突变的动物身上时， 发夹状核酶在结构和功能上保护光感受器 长达8个月。在本提案中，我们描述了扩展这些计划的计划， 通过改进核酶和AAV载体， 通过在ADRP的大型动物模型（转基因猪）中测试治疗; 通过开发不依赖等位基因的核酶来治疗各种视紫红质， 突变;通过采用新的RNA催化剂，以增加潜在的 视紫红质mRNA的靶位点;并通过评估远系小鼠的基因治疗 携带P23 H突变这项工作将得到非侵入性分析的帮助。 技术，使我们能够监测视网膜变性和疗效， 在活体动物中进行治疗。我们希望， 该项目将使我们接近采用AAV载体核酶作为治疗 常染色体显性视网膜色素变性的研究

DESCRIPTION (provided by applicant): The goal of this project is to develop a genetic therapy for autosomal dominant retinitis pigmentosa (ADRP). Retinitis pigmentosa is a type of inherited retinal degeneration caused by the death of rod photoreceptor cells. It affects about 1 person in 3500 and leads to a progressive loss of vision and ultimately blindness, usually over a period of decades. Mutations in over 30 genes lead to retinitis pigmentosa, but defects in the gene for rhodopsin, the major light-harvesting pigment of the rod cell, are the predominant cause of ADRP. These dominant mutations lead to a malfunctioning, mis-sorted, or poorly folded molecule that eventually kills the rod cell that makes it. Our underlying hypothesis is that reducing the expression of these mutated forms of rhodopsin can rescue rod photoreceptors and preserve vision. The genetic tool we will employ for this purpose is a catalytic RNA molecule or ribozyme. Small ribozymes can be engineered to sever almost any RNA in a sequence-specific manner. The instrument we use to introduce the ribozymes into retinal cells is recombinant Adeno-Associated Virus or AAV. During the current funding period, we have designed ribozymes specific for mutant rhodopsin mRNA present in transgenic rat models of ADRP. When delivered to animals bearing the P23H rhodopsin mutation, these hammerhead and hairpin ribozymes protected photoreceptors structurally and functionally for up to 8 months. In this proposal, we describe plans to extend these promising results by improving the ribozymes and the AAV vectors that deliver them; by testing the therapy in a large animal model of ADRP (transgenic pigs); by developing allele-independent ribozymes to treat a variety of rhodopsin mutations; by employing novel RNA catalysts to increase the number of potential target sites in rhodopsin mRNA; and by evaluating gene therapy in outbred mice carrying the P23H mutation. This work will be aided by non-invasive analytical techniques that permit us to monitor retinal degeneration and the efficacy of therapy in living animals. We hope that the successful completion of this project will bring us close to employing AAV-vectored ribozymes as therapy for autosomal dominant retinitis pigmentosa in human patients.