AAV-mediated gene correction in retina

AAV 介导的视网膜基因校正

基本信息

批准号：
7235613
负责人：
JEAN BENNETT
金额：
$ 7.83万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-06-01 至 2008-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7235613
关键词：
ATP phosphohydrolase Academia Adenoviruses Affect Animal Model Animals Bacteria Base Pairing Beta-glucuronidase Binding Biochemical Biological Assay Blindness Cell Death Cell Line Cells Condition DNA DNA annealing Data Dependovirus Disease Electroporation Event Eye Eye diseases Filament Gene Conversion Gene Delivery Gene Mutation Gene Targeting Genes Genetic Genetic Epistasis Genetic Recombination Genetic Transcription Genomics Homologous Gene Immune In Vitro Inherited LacZ Genes Lead Left Mammalian Cell Mediating Methods Microinjections Modification Mus Mutation Numbers Organ Photoreceptors Play Process Production Property Proteins Rate Regulation Reporter Reporting Research Personnel Retina Retinal Retinal Diseases Risk Role Science Single-Stranded DNA Site Structure of retinal pigment epithelium Subfamily lentivirinae Techniques Technology Therapeutic Transfection Transformed Cell Line Transgenic Organisms Update Viral Vector Virus Yeasts adeno-associated viral vector base beta-Galactosidase design gene correction gene therapy homologous recombination in vivo insertion/deletion mutation interest mutant numb protein photoreceptor degeneration promoter repaired research study size therapy development vector viral vector development

项目摘要

DESCRIPTION (provided by applicant): Inherited retinal diseases are a major cause of blindness, and numerous mutations in both retina-specific and nonspecific genes have been identified that cause photoreceptor degeneration. Conventional gene therapy approaches are based on gene addition strategies using viral vectors such as adenovirus (Ad), adeno-associated virus (AAV) and lentivirus. Researchers have demonstrated that virus-mediated gene delivery may be used to successfully delay photoreceptor cell death in animal models of retinal disease; however, the specific endogenous gene mutations are not corrected. Gene targeting allows one to alter genomic sequences at specific chromosomal sites, resulting in permanent gene modifications. Recent reports indicate that AAV can be used to introduce targeted insertions, deletions and substitutions into homologous chromosomal sequences in vitro, with targeting rates that are 4-6 logs higher than those obtained by conventional transfection or electroporation methods. The experiments outlined in this proposal will apply AAV gene-targeting strategies to the retina. The efficacy of virus-mediated repair will be assessed through biochemical, histological and functional assays. Initially studies will be performed in transgenic mutant beta-galactosidase mice to assess if retinal development and viral vector design impact the efficacy of AAV-mediated gene targeting in the retina. Experiments will subsequently be performed in beta- glucuronidase- deficient mice (gusmps/gusmps). These animals will be used to evaluate the therapeutic potential of AAV-mediated gene targeting for RPE disease. The data gathered from these studies may have widespread applications for many inherited pathological conditions and may ultimately lead to the development of therapies that permanently correct fundamental genetic defects responsible for these conditions.

描述（由申请人提供）：遗传性视网膜疾病是失明的主要原因，并且已经鉴定出导致感光受体变性的视网膜特异性和非特异性基因中的许多突变。常规的基因治疗方法基于使用病毒载体（例如腺病毒（AD），腺相关病毒（AAV）和慢病毒的基因添加策略。研究人员表明，病毒介导的基因输送可用于成功延迟视网膜疾病动物模型中的感光细胞死亡。但是，特异性内源基因突变未得到校正。基因靶向允许人们在特定染色体位点改变基因组序列，从而导致永久基因修饰。最近的报告表明，AAV可用于在体外将有针对性的插入，缺失和取代引入同源染色体序列中，其靶向速率比通过常规转染或电孔方法获得的靶向速率高4-6个对数。该提案中概述的实验将对视网膜采用AAV基因靶向策略。病毒介导的修复的功效将通过生化，组织学和功能测定法进行评估。最初将在转基因突变β-半乳糖苷酶小鼠中进行研究，以评估视网膜发育和病毒载体设计是否影响AAV介导的基因靶向视网膜中的功效。实验将随后在β-葡萄糖醛酸酶缺陷的小鼠（GUSMPS/GUSMP）中进行。这些动物将用于评估AAV介导的基因靶向RPE疾病的治疗潜力。从这些研究中收集的数据可能对许多遗传病理状况有广泛的应用，并最终可能导致疗法的发展，这些疗法永久纠正负责这些疾病的基本遗传缺陷。