Novel Genes Expressed in Proliferating Endothelial Cells

眼睛中的新生血管是许多疾病的中心组成部分。导致严重视力障碍的疾病。增殖性糖尿病视网膜病变本身就是导致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在细胞和分子水平上对血管生成的更深入了解。我们研究计划的长期目标是了解当正常静止的眼内皮细胞(EC)去分化为新血管形成所需的增殖、血管生成表型时发生的一系列事件。这项研究集中在我们实验室最近发现的一组新基因，这些基因在增殖的EC中表达。用亲和层析法分离与这些基因相对应的mRNAs(随后克隆)，使用必需的RNA结合蛋白作为诱饵，该蛋白本身在增殖的EC中选择性地表达。我们建议的项目的目标是表征这些新的基因产物，并测试它们是否对血管生成表型至关重要。具体目的#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。在静止和血管生成条件下培养的视网膜内皮细胞将通过免疫化学和杂交技术进行检测，以及体内诱导新生血管的眼组织。在目标#2中，我们将生成全长cdna克隆，以便于结构分析和功能探针(分子和免疫学)的开发。在目标#3中，我们将测试这些分子对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它们抑制增殖和其他表型特有特征的表达的能力来实现。这项研究将增加我们对眼睛增殖性血管内皮细胞基因表达的一般知识，也有可能揭示眼血管生成和其他涉及细胞去分化和增殖的疾病过程中的特定调控点。

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.