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种 常染色体显性遗传性视网膜色素变性(ADRP)的基因治疗视网膜炎 色素性视网膜变性是一种遗传性视网膜变性，由 杆状感光细胞。它每3500人中就有1人感染，并导致 渐进性失明和最终失明，通常在一段时间内 几十年。30多个基因突变导致视网膜色素变性，但缺陷 在视紫红质基因中，视紫红质是视杆细胞的主要捕光色素， 是ADRP的主要原因。这些显性突变导致一种 发生故障、分类错误或折叠不良的分子，最终会杀死 成功的杆状细胞。我们的基本假设是，减少 表达这些突变形式的视紫红质可以挽救杆状感光细胞 并保持视力。为此，我们将使用的遗传工具是 催化RNA分子或核酶。小核酶可以被改造成切割 几乎所有的RNA都是以序列特定的方式。我们用来 重组腺相关核酶导入视网膜细胞 病毒或AAV。在目前的资助期间，我们设计了核酶 在ADRP转基因大鼠模型中存在突变的视紫红质mRNA的特异性。 当传递给携带P23H视紫红质突变的动物时，这些锤头 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功能的计划 通过改进核酶和AAV载体来传递有希望的结果 通过在ADRP(转基因猪)的大型动物模型上测试该疗法； 通过开发等位基因无关的核酶治疗多种视紫红质 突变；通过使用新型RNA催化剂来增加潜在的数量 视紫红质mRNA的靶点；以及通过评估杂交小鼠的基因治疗 携带P23H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退化和疗效 在活体动物上进行治疗。我们希望这项工作的圆满完成 该项目将使我们接近使用AAV载体核酶作为治疗 人类患者的常染色体显性遗传性视网膜色素变性。

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.