Use Of Cdna Microarrays In Gene Expression Of Uveitis Pa

CDNA 微阵列在葡萄膜炎基因表达中的应用

• 批准号: 6826917
• 负责人: ROBERT B. NUSSENBLATT
• 金额: --
• 依托单位: NATIONAL EYE INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6826917
• 关键词: T lymphocyte; complementary DNA; cytokine; family genetics; gene expression; gene mutation; genetic markers; genetic susceptibility; genome; human genetic material tag; human subject; human tissue; inflammation; leukocyte activation /transformation; linkage mapping; microarray technology; myopia; nucleic acid sequence; polymerase chain reaction; restriction fragment length polymorphism; single strand conformation polymorphism; tumor necrosis factor alpha; uveitis

Ocular inflammatory diseases, including uveitis, cause significant visual loss. Previous non-human investigations have identified several cell types, receptor systems and metabolic intermediates that have led to treatment approaches for human patients. However,information on the human genetic expression of these steps in defined inflammatory disease states is lacking. This non-intervention study proposes to obtain peripheral blood and tissue specimens from patients enrolled in other intramural trials for ocular inflammatory diseases and to apply contemporary cDNA microarray technologies for the analysis of differential gene expression. Test results will not be reported to participants or used for diagnostic or therapeutic purposes.The study?s primary objective is to identify unique gene expression profiles as well as disease relevant genes for patients with ocular inflammatory disease at defined clinical stages using cDNA microarray analysis. This will help provide further insight to understand the pathological mechanisms and potential targets for treatment. Some 3,000-5,000 genes will be examined starting with a selected set associated with interleukin (IL) proteins and their receptors, and with tumor necrosis factors (TNF). Purified peripheral blood mononuclear cells (or whole blood lysates using RNA isolation procedures) will be used to isolate total RNA from these samples. Samples will be taken during periods of active or recurring inflammatory disease and again during periods of quiescence after treatment. The microarray tools and methods for genetic analysis are now available at the NEI.In conjunction with these studies are functional assays and cell surface markers of negative regulatory cells, so called ?suppressor cells?. These studies center for the moment on the CD25+ T cell fraction and certain markers of T cell activation such as GITR. A correlation is being made between T cell activation and these negative regulatory signals and clinical activity.

包括葡萄膜炎在内的眼部炎症性疾病会导致严重的视力丧失。以前的非人类研究已经确定了几种细胞类型，受体系统和代谢中间体，这些细胞类型，受体系统和代谢中间体导致了人类患者的治疗方法。然而，在确定的炎症性疾病状态下，这些步骤的人类基因表达的信息是缺乏的。这项非干预性研究建议从其他眼炎性疾病的壁内试验中招募的患者中获得外周血和组织标本，并应用当代cDNA微阵列技术分析差异基因表达。测试结果将不会报告给参与者或用于诊断或治疗目的。的主要目的是确定独特的基因表达谱，以及疾病相关基因的眼部炎症性疾病患者在确定的临床阶段，使用cDNA微阵列分析。这将有助于进一步了解病理机制和潜在的治疗靶点。大约3000 - 5000个基因将被检查，从与白细胞介素（IL）蛋白及其受体以及与肿瘤坏死因子（TNF）相关的选定集合开始。纯化的外周血单核细胞（或使用RNA分离程序的全血裂解物）将用于从这些样本中分离总RNA。将在活动性或复发性炎性疾病期间采集样本，并在治疗后静止期再次采集样本。微阵列工具和遗传分析方法现在可以在NEI获得。与这些研究相结合的是负调节细胞的功能测定和细胞表面标记，所谓的？抑制细胞？这些研究目前集中在CD 25 + T细胞部分和T细胞活化的某些标志物如GITR上。T细胞活化与这些负调控信号和临床活性之间存在相关性。