RDS mutations: Gene therapy for ADRP, macular degeneration and pattern dystrophy

描述（由申请人提供）：本研究的长期目标是为导致失明的主要遗传性视网膜疾病提供基因治疗。该项目的目的是开发一种治疗RDS/外周蛋白基因突变的方法，这种基因突变导致模式营养不良、黄斑变性和常染色体显性视网膜色素变性（ADRP）。在这项工作中，我们将使用含有RDS/外周蛋白基因突变的小鼠模型，我们将使用腺相关病毒来传递潜在的治疗基因。由于RDS/外周蛋白基因的突变经常与视锥细胞的变性有关，我们将把治疗基因的表达定位于视锥细胞和视杆细胞。我们有四个具体目标：(1)开发一种RDS/peripherin的RNA替代技术，其中使用核酶或siRNA降低内源性mRNA的水平，同时提供核酶或siRNA抗性的mRNA版本。(2)在表达RDS/peripherin突变版本导致ADRP的P216L小鼠系上测试基因替代方法。(3)建立受外周蛋白/rd突变影响的斑状营养不良和黄斑变性的视锥细胞的AAV介导基因靶向方法。(4)在NRL基因敲除仅产生锥体光感受器的小鼠中表达P216L突变，并在该模型中检测RNA替代。我们的实验室和其他研究人员的初步结果表明，这些实验将证明是富有成效的，并将导致治疗由RDS突变引起的视网膜疾病。

DESCRIPTION (provided by applicant): The long-term goal of this research is to generate a gene therapy for dominantly inherited forms of retinal disease which lead to blindness. The aims of this project are designed to develop a treatment for mutations in the RDS/peripherin gene which lead to pattern dystrophy, macular degeneration and autosomal dominant retinitis pigmentosa (ADRP). For this work we will employ mouse models that contain mutations in the RDS/peripherin gene, and we will use Adeno-associated virus to deliver potential therapeutic genes. Because mutations in the RDS/peripherin gene are frequently associated with degeneration of cone photoreceptor cells, we will target the expression of therapeutic genes to cones in addition to rod cells. We have four specific aims: (1) To develop an RNA replacement technology for RDS/peripherin in which levels of endogenous mRNA are reduced using a ribozyme or an siRNA, and a ribozyme- or siRNA-resistant version of the mRNA is supplied simultaneously. (2) To test the gene replacement method in the P216L mouse line that expresses a mutant version of RDS/peripherin leading to ADRP. (3) To develop an AAV- mediated gene targeting method for cone photoreceptor cells, the cells affected by peripherin/rd mutations in pattern dystrophy and macular degeneration. (4) To express the P216L mutation in the context of the NRL knockout mouse, which produces only cone photoreceptors, and to test RNA replacement in this model. Preliminary results from our laboratories and from those of other investigators suggest that these experiments will prove fruitful and will lead to a treatment for retinal diseases caused by RDS mutations. This project is relevant to age-related macular degeneration and retinitis pigmentosa. Age-related macular degeneration (AMD) is the major blinding disease affecting the elderly in developed countries. Some form of AMD affects as many as 1 in 3 people over the age of 70. It leads to scarring and degeneration of the cone-rich central retina, which is required for acute vision, including reading and face-recognition. Retinitis Pigmentosa (RP) affects fewer people, 60,000-100,000 in the US, but it robs them of useful vision at an earlier age. Since many of the mutant genes for RP are known, a gene therapy may be possible for many forms of the disease.