Genetic Modulation of Glaucoma

青光眼的基因调控

• 批准号: 8269646
• 负责人: MONICA M JABLONSKI
• 金额: $ 37.49万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8269646
• 关键词: Alleles; Atrophic; Bioinformatics; Biological Assay; Blindness; Candidate Disease Gene; Cell Count; Clinical; Data; Data Set; Disease; Disease model; Evaluation; Eye; Eye diseases; Genes; Genetic; Genotype; Glaucoma; Haplotypes; Human; Human Chromosomes; Investigation; Iris; Iris Diseases; Laboratories; Lead; Link; Literature; Maps; Methods; Microarray Analysis; Molecular; Molecular Models; Mus; Mutation; Open-Angle Glaucoma; Optic Nerve; Outcome; Parents; Patients; Penetrance; Phenotype; Physiologic Intraocular Pressure; Pigments; Predisposition; Quantitative Trait Loci; Reading; Recombinant Inbred Strain; Research; Research Personnel; Resistance; Retina; Severities; Severity of illness; Statistical Models; Surveys; Testing; Translations; Variant; Whole-Genome Shotgun Sequencing; Work; age related; authority; cohort; expectation; experience; ganglion cell; molecular modeling; mutant; trait

In this proposal, we use our enlarged set of BXD recombinant inbred strains to identify gene loci that are involved in modulating the severity of glaucoma. Our approach combines a thorough clinical and laboratory examination, and microarray analysis of the entire set of 81 BXD lines generated over the last 10 years with the express purpose of studying the genetics of eye disease and glaucoma. One of the parental strains of BXD, DBA/2J, develop an age-related glaucoma that is preceded by iris atrophy and pigment dispersion. While mutant alleles of two genes, Tryp1 and Gpnmb, cause the iris disease in D2, the literature strongly suggests that these mutations are not sufficient to cause glaucoma. Specifically, introgression of both mutant alleles onto C57BL/6J, the other parent strain of BXD, results in a marked resistance to optic nerve damage. This indicates that genes other than those that cause pigment dispersion influence the glaucomatous phenotype. The current proposal describes a unique opportunity to define the modifying loci. In Aim 1, we test the hypothesis that the combined mutations in Tryp1 and Gpnmb are not sufficient to cause all aspects of the glaucoma phenotype. If our hypothesis is true, we expect to see that the severity of disease is not solely dependent on the two mutant alleles. We have already identified multiple BXD strains in which IOP and genetic diplotype are not correlated. As we systematically examine all BXD strains and establish relations between phenotype and diplotype, we fully expect to find additional strains that defy expectations of a simple two-locus disease model. In Aim 2, we define loci and genes that modulate glaucoma severity. To do so, we will identify and evaluate candidate genes within loci that modulate the severity of the glaucoma phenotype. We will exploit our new whole genome shotgun sequence for D2 (about >50x short read coverage generated at UTHSC in 2009) along with massive whole eye and whole retina expression datasets that we have also generated as a prelude to this work. We expect to efficiently nominate and evaluate candidate glaucoma genes using state-of-the-art bioinformatic methods and conventional molecular assays. In Aim 3, we use bidirectional translation. We test the translational validity of mouse candidates from Aim 2 using cohorts of human glaucoma patients. Dr. J. Wiggs and colleagues will perform focused gene association studies using candidate glaucoma genes nominated in Aim 2. Specifically, we use association analyses of markers encompassing syntenic regions of human chromosomes. In reciprocal reverse translation, we (MMJ and LL) will evaluate known, new, and candidate glaucoma genes from clinical cohorts and determine if and how these variants are associated with glaucoma- associated traits in BXDs. Combining the top priority gene candidates from both mouse and human glaucoma studies, we will generate molecular and statistical models of susceptibility candidate genes, linked phenotypes, and associated mechanisms.

在此提案中，我们使用扩大的BXD重组菌株集来鉴定基因基因座 参与调节青光眼的严重程度。我们的方法结合了一个彻底的临床和实验室 检查和微阵列分析对过去10年中生成的整个81种BXD线的分析 研究眼病和青光眼的遗传学的明确目的。 BXD的父母菌株之一， DBA/2J发展与年龄相关的青光眼，此前是虹膜萎缩和色素分散体。尽管 两个基因的突变等位基因TRYP1和GPNMB引起D2中的虹膜疾病，文献强烈表明 这些突变不足以引起青光眼。具体而言，两个突变等位基因的渗入 C57BL/6J是BXD的另一个父菌株，导致对视神经损伤的明显抗性。这表明 除引起色素色散的基因以外的基因会影响青光眼表型。电流 提案描述了定义修改基因座的独特机会。在AIM 1中，我们检验了以下假设 Tryp1和GPNMB中的合并突变不足以引起青光眼表型的各个方面。如果 我们的假设是正确的，我们希望看到疾病的严重程度不仅取决于两个突变体 等位基因。我们已经确定了多个BXD菌株，其中IOP和遗传外交类型与无关。 当我们系统地检查所有BXD菌株并建立表型与外交类型之间的关系时，我们 完全期望找到其他菌株，反对对简单的两局疾病模型的期望。在AIM 2中，我们 定义调节青光眼严重程度的基因座和基因。为此，我们将确定和评估候选人 基因座内的基因调节青光眼表型的严重程度。我们将利用我们的新整体 D2的基因组shot弹枪序列（2009年在UTHSC上产生的大约50倍的短读覆盖范围）以及 我们也为这项工作的前奏而生成的全眼和全视网膜表达数据集。 我们希望使用最先进的生物学有效提名和评估候选青光眼基因 方法和常规分子测定。在AIM 3中，我们使用双向翻译。我们测试 使用人青光眼患者的同类群体，来自AIM 2的小鼠候选者的翻译有效性。 J. Wiggs博士 同事将使用提名的候选青光眼基因进行重点的基因关联研究 目的2。具体来说，我们使用涵盖人类同义区域的标记的关联分析 染色体。在相互反向翻译中，我们（MMJ和LL）将评估已知，新的和候选人 来自临床队列的青光眼基因，并确定这些变体是否以及如何与青光眼相关 BXD中的相关性状。结合小鼠和人青光眼的最高优先基因候选者 研究，我们将生成连接的敏感性候选基因的分子和统计模型 表型和相关机制。