Comparative Genomics of Glaucoma

青光眼的比较基因组学

• 批准号: 6918142
• 负责人: Robert R. H Anholt
• 金额: $ 43.47万
• 依托单位: NORTH CAROLINA STATE UNIVERSITY RALEIGH
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6918142
• 关键词: African; Drosophilidae; RNA interference; arthropod genetics; comparative genomic hybridization; gene environment interaction; gene expression; gene targeting; genetic polymorphism; genetic screening; genetic susceptibility; genetically modified animals; glaucoma; human population genetics; human subject; microarray technology; molecular genetics; northern blottings; phenotype; polymerase chain reaction; protein protein interaction; receptor expression; transposon /insertion element; vision tests

DESCRIPTION (provided by applicant): Our long-term goal is to identify ensembles of genes that undergo altered transcriptional regulation during the early stages of the development of some forms of glaucoma, and to identify in the human population at large polymorphisms in such genes that may predispose individuals who harbor them to the development of the disease. Glaucoma is the leading treatable cause of irreversible blindness worldwide and affects more than 60 million people. The genetic factors that predispose to glaucoma in the population at large remain, however, largely unknown. Previously, we established Drosophila melanogaster as a model system for studies of the effects of the human glaucoma-associated "trabecular meshwork inducible glucocorticoid response" protein (also known as myocilin; TIGR/MYOC) and identified transcripts that are up- or down-regulated when TIGR/MYOC is overexpressed in the Drosophila eye. Drosophila remains unsurpassed as a versatile genetic model, which can be used as a rapid gene discovery system, in which transcriptional profiling can identify genes that undergo altered regulation when candidate glaucoma genes are expressed in the Drosophila eye. After identifying human orthologues of such genes we can ask which of them show alterations in their expression in perfused post mortem human eyes when TIGR/MYOC (or other candidate glaucoma genes) are introduced via an adenovirus. We will then investigate whether polymorphisms in candidate genes implicated via this two-step screen are linked to the incidence of glaucoma in a West-African human population. The specific aims of this proposal are to: (l)'Assess the ocular phenotypes of flies with targeted overexpression of TIGR/MYOC mutants and targeted RNAi mediated downregulation of TIGR/MYOC-modulated genes and perform analyses and comparisons of transcriptional alterations among transgenic strains; (2) Assess whether human homologues of Drosophila genes, discovered previously and in Specific Aim 1, are up- or down-regulated in perfused post mortem human eyes subjected to overexpression of TIGR/MYOC, or of other new candidate glaucoma genes; and, (3) Evaluate whether polymorphisms in new candidate glaucoma genes are associated with ocular hypertension and the incidence of glaucoma in a West-African population with high incidence of the disease. These experiments represent a novel strategy for candidate disease susceptibility gene discovery and will lay the foundation for future large scale genomic SNP studies to identify the full complement of genes that may convey susceptibility to glaucoma.

描述（由申请人提供）：我们的长期目标是鉴定在某些形式的青光眼发展的早期阶段经历改变的转录调控的基因群，并在人群中鉴定这些基因的大量多态性，这些多态性可能使携带这些基因的个体容易患上这种疾病。青光眼是全球导致不可逆失明的主要可治疗原因，影响超过 6000 万人。然而，在广大人群中易患青光眼的遗传因素仍然很大程度上未知。此前，我们建立了果蝇作为模型系统，用于研究人类青光眼相关的“小梁网诱导糖皮质激素反应”蛋白（也称为肌纤蛋白；TIGR/MYOC）的影响，并鉴定了当 TIGR/MYOC 在果蝇眼中过表达时上调或下调的转录本。果蝇作为一种多功能遗传模型仍然是无与伦比的，它可以用作快速基因发现系统，其中转录分析可以识别当候选青光眼基因在果蝇眼睛中表达时经历改变调节的基因。在鉴定出这些基因的人类直向同源物后，我们可以询问当通过腺病毒引入 TIGR/MYOC（或其他候选青光眼基因）时，其中哪些基因在灌注的死后人眼中表现出表达变化。然后，我们将研究通过两步筛选涉及的候选基因的多态性是否与西非人群青光眼的发病率相关。该提案的具体目标是： (l) 通过靶向过度表达 TIGR/MYOC 突变体和靶向 RNAi 介导的 TIGR/MYOC 调节基因下调来评估果蝇的眼部表型，并对转基因品系之间的转录改变进行分析和比较； (2) 评估之前在具体目标 1 中发现的果蝇基因的人类同源物在经过 TIGR/MYOC 或其他新候选青光眼基因过表达的死后人眼灌注中是否上调或下调； (3) 评估新候选青光眼基因的多态性是否与高眼压症和青光眼高发西非人群的发病率相关。这些实验代表了一种发现候选疾病易感基因的新策略，并将为未来大规模基因组 SNP 研究奠定基础，以确定可能传达青光眼易感性的全部基因。