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显性遗传形式的视网膜疾病产生基因疗法。该项目的目的是开发一种治疗RDS/peripherin基因突变的方法，这些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，我们将采用含有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光感受器细胞的变性相关，因此我们将治疗基因的表达靶向于视锥细胞而非视杆细胞。我们有四个具体目标：（1）开发RDS/外周蛋白的RNA置换技术，其中使用核酶或siRNA降低内源性mRNA的水平，并同时提供核酶或siRNA抗性形式的mRNA。(2)在表达导致ADRP的RDS/外周蛋白突变体的P216 L小鼠系中检测基因置换方法。(3)目的：建立一种腺相关病毒介导的视锥细胞基因打靶方法，用于治疗视网膜色素变性和黄斑变性中外周蛋白/rd突变的视锥细胞。(4)在仅产生视锥光感受器的NRL敲除小鼠的背景下表达P216 L突变，并在该模型中测试RNA置换。我们实验室和其他研究人员的初步结果表明，这些实验将证明是富有成效的，并将导致对RDS突变引起的视网膜疾病的治疗。 本项目与老年性黄斑变性和视网膜色素变性有关。视网膜相关性黄斑变性（AMD）是影响发达国家老年人的主要致盲性疾病。某种形式的AMD影响多达三分之一的70岁以上的人。它会导致富含视锥细胞的中央视网膜的瘢痕形成和退化，而这是包括阅读和面部识别在内的敏锐视觉所必需的。视网膜色素变性（RP）影响的人较少，在美国有60，000 - 100，000人，但它在早期剥夺了他们的有用视力。由于已知许多RP的突变基因，因此基因治疗可能用于许多形式的疾病。

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.