GENE EXPRESSION CHANGES IN GLAUCOMA

青光眼的基因表达变化

• 批准号: 6616732
• 负责人: RONALD H FARKAS
• 金额: $ 18.04万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-08-01 至 2005-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6616732
• 关键词: cell death; flow cytometry; gene expression; glaucoma; immunocytochemistry; in situ hybridization; laboratory mouse; molecular cloning; molecular pathology; northern blottings; nucleic acid sequence; retinal ganglion; serial analysis of gene expression; western blottings

Glaucoma is the second leading cause of blindness worldwide, and is especially prevalent among African Americans. There are many different types of glaucoma, but all share the common defining feature of retinal ganglion cell death. The overall goal of this proposal is to use recently developed techniques to understand better the molecular events underlying the death of retinal ganglion cells in glaucoma. This will likely lead to new treatments for glaucoma and other diseases of the optic nerve. My working hypothesis is that intraocular pressure, as well as other as yet unknown insults, leads to changes in gene expression that characterize the ganglion cell response to injury, from initial insult to induction of cell death mechanisms. The long- term goal is to understand the function of these genes so that they can be therapeutically modulated in glaucoma to prevent or delay retinal ganglion cell death. The murine glaucoma model is proposed as a system for analysis, for reasons that are described in the application. The first specific aim is to identify genes that are differentially expressed in the retina of mice with "glaucoma." To accomplish this, the technique of 'serial analysis of gene expression' (SAGE) will be applied to whole retinal tissue. Genes that demonstrate significant differences in expression level will be selected for detailed study. A key advantage of this approach is that it can identify, free of preconceived bias, previously unsuspected genes that may be important in glaucoma, as opposed to a 'candidate gene' approach that might focus, for example, solely on genes related to apoptosis or growth factors. The second specific aim is to isolate pure retinal ganglion cells for use in similar gene expression studies. The use of purified ganglion cells will allow for high-resolution investigation of gene expression changes specific to ganglion cells. To accomplish these aims, technologies for gene expression analysis will be refined and developed. A combination of research and didactic activities will contribute to the training value of the proposal by focusing both on currently achievable scientific ends and on the skills needed by the primary investigator for a future independent research career. The Wilmer Ophthalmological Institute at Johns Hopkins University, and the sponsor's laboratory in particular, provide an ideal environment for the candidate to pursue these goals.

青光眼是全球失明的第二大主要原因，在非裔美国人中尤其普遍。 青光眼有许多不同类型的类型，但所有人都具有视网膜神经节细胞死亡的共同定义特征。 该提案的总体目标是使用最近开发的技术来更好地了解视网膜神经节细胞在青光眼中死亡的分子事件。 这可能会导致针对视神经的青光眼和其他疾病的新疗法。我的工作假设是，眼内压以及其他未知的侮辱会导致基因表达的变化，从初始侮辱到诱导细胞死亡机制，这表征了神经节细胞对损伤的反应。 长期目标是了解这些基因的功能，以便可以在青光眼中调节它们，以预防或延迟视网膜神经节细胞死亡。 出于应用程序中描述的原因，提出了鼠青光眼模型作为分析系统。 第一个具体目的是鉴定出在小鼠的视网膜中差异表达的基因。 为此，“基因表达的序列分析”（SAGE）的技术将应用于整个视网膜组织。将选择表现出表达水平显着差异的基因进行详细研究。 这种方法的一个关键优点是，它可以识别出可能在青光眼中可能很重要的先前未经疑问的偏见，而不是与“候选基因”方法相反，而“候选基因”方法可能仅仅集中在与凋亡或生长因子有关的基因上。 第二个特定目的是分离纯净视网膜神经节细胞，以用于相似的基因表达研究。 纯化的神经节细胞的使用将允许对神经节细胞的基因表达变化进行高分辨率研究。 为了实现这些目标，将完善和开发用于基因表达分析的技术。 研究和教学活动的结合将促进该提案的培训价值，通过将重点放在目前可实现的科学目的以及主要研究人员为未来独立研究职业所需的技能上。 约翰·霍普金斯大学（Johns Hopkins University）的威尔默眼科研究所，特别是赞助商的实验室，为候选人提供了追求这些目标的理想环境。