Novel Genes Expressed in Proliferating Endothelial Cells

增殖内皮细胞中表达的新基因

• 批准号: 6895082
• 负责人: MARK Collin ALLIEGRO
• 金额: $ 21.3万
• 依托单位: LSU HEALTH SCIENCES CENTER
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-05-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6895082
• 关键词: angiogenesis; cell cycle; cell differentiation; cell migration; cell proliferation; complementary DNA; confocal scanning microscopy; electron microscopy; eye; gene expression; gene targeting; immunocytochemistry; immunoelectron microscopy; in situ hybridization; laboratory mouse; laboratory rabbit; messenger RNA; molecular cloning; monoclonal antibody; phenotype; vascular endothelium

Neovascularization in the eye is a central component of many. disorders leading to severe vision impairment. Proliferative diabetic retinopathy alone is the leading cause of blindness in Americans aged 20-74. Although significant progress has been made, our ability to prevent and treat neovascular disorders would clearly benefit from a more thorough knowledge of angiogenesis at the cellular and molecular levels. The long term goal of our research program has been to understand the sequence of events that occur when normally quiescent ocular endothelial cells (EC) dedifferentiate into the proliferative, angiogenic phenotype required for new blood vessel formation. This study focuses on a set of novel genes recently identified in our laboratory that are expressed in proliferating EC. mRNAs corresponding to these genes were isolated by affinity chromatography (and subsequently cloned) using an essential RNA binding protein as bait that is itself expressed selectively in proliferating EC. Our aims for the proposed project are to characterize these novel gene products and test whether they are essential to the angiogenic phenotype. Specific Aim #1 is designed to tell us which are upregulated in proliferating EC, and therefore more likely to function in promoting or maintaining the angiogenic phenotype. Cultured retinal EC under quiescent and angiogenic conditions will be examined by immunochemical and hybridization techniques, as will be ocular tissues induced to undergo neovascularization in vivo. In Aim #2, we will generate full length cDNA clones to facilitate structural analysis and development of functional probes (molecular and immunological). In Aim #3 we will test the hypothesis that these molecules are essential for maintaining the angiogenic phenotype. This will be accomplished by introducing antagonists into cells and assessing their ability to inhibit proliferation and expression of other phenotype-specific traits. This study will add to our general knowledge of gene expression in proliferating vascular EC of the eye, but also has the potential for revealing specific regulatory points in ocular angiogenesis and other disease processes involving cell dedifferentiation and proliferation.

眼睛的新生血管形成是许多疾病的核心组成部分。导致严重视力障碍的疾病。仅增殖性糖尿病视网膜病变就是 20-74 岁美国人失明的主要原因。尽管已经取得了重大进展，但我们预防和治疗新生血管疾病的能力显然将受益于对细胞和分子水平上血管生成的更全面了解。我们研究计划的长期目标是了解当通常静止的眼内皮细胞 (EC) 去分化为新血管形成所需的增殖性血管生成表型时发生的事件顺序。本研究重点关注我们实验室最近发现的一组在增殖的 EC 中表达的新基因。 使用必需的RNA结合蛋白作为诱饵，通过亲和层析分离（并随后克隆）对应于这些基因的mRNA，该诱饵本身在增殖的EC中选择性表达。我们拟议项目的目标是表征这些新基因产物并测试它们是否对血管生成表型至关重要。具体目标#1 旨在告诉我们哪些在增殖的 EC 中上调，因此更有可能在促进或维持血管生成表型方面发挥作用。将通过免疫化学和杂交技术检查在静止和血管生成条件下培养的视网膜EC，以及诱导眼组织在体内经历新血管形成。在目标 #2 中，我们将生成全长 cDNA 克隆，以促进结构分析和功能探针（分子和免疫学）的开发。在目标#3 中，我们将检验这些分子对于维持血管生成表型至关重要的假设。这将通过将拮抗剂引入细胞并评估其抑制增殖和其他表型特异性特征表达的能力来实现。这项研究将增加我们对眼部血管内皮细胞增殖中基因表达的一般了解，而且还有可能揭示眼部血管生成和涉及细胞去分化和增殖的其他疾病过程中的特定调节点。