Models of X-Linked Retinitis Pigmentosa

X连锁色素性视网膜炎模型

• 批准号: 7277187
• 负责人: GUSTAVO David AGUIRRE
• 金额: $ 38.48万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-07-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7277187
• 关键词: Accounting; Affect; Alleles; Animal Model; Animals; Antibodies; Benign; Biochemical; Birds; C-terminal; Canis familiaris; Catalytic RNA; Cells; Charge; Code; Complementary DNA; Crossbreeding; Custom; Cyclic GMP; Development; Disease; Dominant Genetic Conditions; Early Intervention; Environmental Risk Factor; Evaluation; Exons; Frameshift Mutation; Frequencies; Gel; Gene Expression; Gene Family; Gene Mutation; Gene Proteins; Genes; Genetic Recombination; Genome Scan; Genomics; Goals; Grant; Guanosine Triphosphate Phosphohydrolases; Haplotypes; Heterogeneity; Hot Spot; Human; Introns; Investigational Therapies; Length; Link; Maps; Mediating; Messenger RNA; Metabolic; Modeling; Molecular; Molecular Profiling; Mutation; Nature; Night Blindness; Oligonucleotides; Pathogenesis; Pathology; Patients; Personal Communication; Phenotype; Photoreceptors; Physical Map of the Human Genome; Population; Population Study; Principal Investigator; Process; Protein Isoforms; Proteins; Purines; RNase protection assay; Range; Regulator Genes; Resources; Retina; Retinal; Retinal Degeneration; Retinitis Pigmentosa; Role; Severities; Severity of illness; Staging; Testing; Therapeutic Intervention; Time; Tissues; Transcript; Variant; Visual; Visual Fields; base; cDNA Arrays; developmental disease; disease phenotype; disease-causing mutation; early onset; founder mutation; gain of function; gene replacement; gene therapy; genetic pedigree; insight; loss of function; male; microdeletion; mutant; novel; photoreceptor degeneration; physical mapping; programs; protein aminoacid sequence; purine; retinal rods; vector

DESCRIPTION (provided by applicant): Mutations in the RP GTPase regulator gene (RPGR) cause the RP3 form of X-linked RP, one of the most prevalent and severe forms of retinitis pigmentosa. Over half of these mutations occur in ORF15, a purine rich repetitive domain of unknown function. During the current grant we fine mapped two X-linked canine retinal degenerations (XLPRA1 and 2), and found 2 different causative rnicrodeletions in ORF15. These are the only animal models for ORF15-associated disease. Because the 2 disorders are phenotypically very distinct, the major differences between them reflect the nature of the RPGRORF15 microdeletions. Within XLPRA1 however, although all affected dogs share a common stable mutation originating from a single founder, significant variation in disease severity exists. This clearly results not from environmental factors or heterogeneity at the primary locus, but as a semi-dominant trait segregating independently of the primary disease locus. SNP-based haplotypes will be developed to test genes known to interact with RPGR, as candidate disease modifiers. Informative pedigrees will be examined for cosegregation with disease severity. If no such association is found, a genome scan will then be undertaken to define genomic regions with potential positional candidates. We will exploit differences in expression, at both the mRNA and protein level, to identify disease-associated transcripts and gene products. RNase protection assays will be used to identify ORF15 containing transcripts and evaluate expression in retinal vs. nonretinal tissues. Because both mutations alter the length and charge of the ORF15 proteins, 2D-gels and peptide sequencing will be used to identify disease relevant isoforms. Specific antibodies will be developed for immunochemical studies to evaluate expression and localization of each isoform during development and degeneration. Using our custom canine retinal cDNA microarray we will identify changes in gene expression that are either mutation specific or common to both disorders. This will reveal insights into the molecular processes underlying the disease and degeneration of visual cells. Further, it will help to indicate when irreversible metabolic changes occur that could limit therapeutic intervention, and establish time points for implementation and evaluation of our planned gene therapy using AAV 2/5 pseudotyped vectors. Two hypotheses will be tested experimentally with gene therapy: a) -that disease from the stop mutation (XLPRA1) is caused by loss of function requiring gene replacement; and b) -that disease from the frameshift mutation (XLPRA2) arises from a deleterious gain of function, necessitating early intervention with combined ribozyme knockdown and gene replacement.

描述（由申请人提供）：RP GTPase调控基因（RPGR）突变导致RP3形式的x -连锁RP，这是色素性视网膜炎最常见和最严重的形式之一。超过一半的突变发生在ORF15，一个功能未知的富含嘌呤的重复结构域。在目前的资助中，我们精细地绘制了两个x连锁犬视网膜变性（XLPRA1和2），并在ORF15中发现了2个不同的致病性微缺失。这些是orf15相关疾病的唯一动物模型。由于这两种疾病在表型上非常不同，它们之间的主要差异反映了rpgorf15微缺失的性质。然而，在XLPRA1中，尽管所有受影响的狗都有一个共同的稳定突变，源自单一的创始人，但疾病严重程度存在显着差异。这显然不是由环境因素或原发位点的异质性造成的，而是作为一种独立于原发疾病位点分离的半显性性状。基于snp的单倍型将用于测试已知与RPGR相互作用的基因，作为候选疾病修饰因子。信息性家系将被检查是否与疾病严重程度共分离。如果没有发现这样的关联，那么将进行基因组扫描，以确定具有潜在位置候选的基因组区域。