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

描述(申请人提供)：这项研究的长期目标是为导致失明的主要遗传性视网膜疾病产生一种基因疗法。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，该基因突变会导致模式营养不良、黄斑变性和常染色体显性遗传性视网膜色素变性(Adrp)。在这项工作中，我们将使用包含RDS/外周蛋白基因突变的小鼠模型，并使用腺相关病毒来传递潜在的治疗基因。由于RDS/外周蛋白基因突变经常与视锥感光细胞的退化有关，我们将针对视锥细胞和视杆细胞之外的治疗基因的表达。我们有四个具体的目标：(1)开发一种针对RDS/外周蛋白的RNA替代技术，其中使用核酶或siRNA来降低内源mRNA的水平，同时提供核酶或siRNA抗性版本的mRNA。(2)在表达突变型RDS/外周蛋白导致adrp的P216L小鼠系中检测基因置换方法。(3)建立AAV介导的视锥感光细胞基因打靶方法，研究视锥感光细胞受外周蛋白/RD突变影响的模式营养不良和黄斑变性细胞。(4)在仅产生锥状光感受器的NRL基因敲除小鼠中表达P216L突变，并在该模型中检测RNA替换。我们实验室和其他研究人员的初步结果表明，这些实验将被证明是富有成效的，并将导致治疗由RDS突变引起的视网膜疾病。 该项目与老年性黄斑变性和视网膜色素变性有关。老年性黄斑变性(AMD)是发达国家影响老年人的主要致盲疾病。某种形式的AMD影响多达三分之一的70岁以上的人。它会导致富含视锥的中央视网膜的疤痕和退化，这是敏锐视觉所必需的，包括阅读和面部识别。视网膜色素变性(RP)影响的人数较少，在美国为60,000-100,000人，但它在较早的年龄就剥夺了他们有用的视力。由于RP的许多突变基因是已知的，因此基因疗法可能适用于多种形式的疾病。

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.