Genetic Modulation of Glaucoma

青光眼的基因调控

• 批准号: 8473870
• 负责人: MONICA M JABLONSKI
• 金额: $ 35.63万
• 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/8473870
• 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.

在本提案中，我们使用扩大的 BXD 重组自交系菌株来鉴定以下基因位点： 参与调节青光眼的严重程度。我们的方法结合了彻底的临床和实验室 对过去 10 年中生成的整套 81 个 BXD 品系进行检查和微阵列分析 表达研究眼病和青光眼遗传学的目的。 BXD 亲本菌株之一， DBA/2J，出现与年龄相关的青光眼，随后出现虹膜萎缩和色素分散。尽管 两个基因 Tryp1 和 Gpnmb 的突变等位基因导致 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。具体来说，两个突变等位基因渗入到 BXD 的另一个亲本菌株 C57BL/6J 对视神经损伤具有显着的抵抗力。这表明 除了导致色素分散的基因之外，还有其他基因影响青光眼表型。目前的 提案描述了定义修饰基因座的独特机会。在目标 1 中，我们检验以下假设： Tryp1 和 Gpnmb 的组合突变不足以引起青光眼表型的所有方面。如果 我们的假设是正确的，我们期望看到疾病的严重程度不仅仅取决于这两种突变体 等位基因。我们已经鉴定了多个 BXD 菌株，其中 IOP 和遗传双倍型不相关。 当我们系统地检查所有 BXD 菌株并建立表型和双倍型之间的关系时，我们 完全期望找到其他菌株，这些菌株违背了简单的双基因座疾病模型的预期。在目标 2 中，我们 定义调节青光眼严重程度的基因座和基因。为此，我们将识别并评估候选人 位点内调节青光眼表型严重程度的基因。我们将利用我们的新整体 D2 的基因组鸟枪序列（约 50 倍于 2009 年 UTHSC 生成的短读覆盖率）以及 我们还生成了大量的全眼和全视网膜表达数据集，作为这项工作的前奏。 我们期望使用最先进的生物信息学有效地提名和评估候选青光眼基因 方法和常规分子测定。在目标 3 中，我们使用双向翻译。我们测试了 使用人类青光眼患者队列研究来自 Aim 2 的小鼠候选者的翻译有效性。 J·威格斯博士 和同事将使用提名的候选青光眼基因进行重点基因关联研究 目标 2. 具体来说，我们使用包含人类同线性区域的标记的关联分析 染色体。在相互逆向翻译中，我们（MMJ 和 LL）将评估已知的、新的和候选的 来自临床队列的青光眼基因，并确定这些变异是否以及如何与青光眼相关 BXD 中的相关特征。结合小鼠和人类青光眼的最优先候选基因 研究，我们将生成易感性候选基因的分子和统计模型，将其链接起来 表型和相关机制。