REGULATION OF VEGF MRNA STABILITY BY HYPOXIA

说明(改编自《调查者摘要》)： 血管内皮生长因子(VEGF)是一种重要的血管内皮细胞生长因子 糖尿病视网膜病变、癌症和冠心病的血管生成。 血管内皮生长因子的表达受细胞内氧的严格控制 紧张。以往的研究表明，血管内皮生长因子mRNA显著地 在低氧条件下稳定。潜在的分子机制 对血管内皮细胞生长因子mRNA稳定性的调控还不是很清楚。在 初步应用结果表明，血管内皮生长因子信使核糖核酸的稳定性 由其3‘非编码区中的顺式作用元件控制。他们还拥有 确定了与该元件结合的反式作用因子(HUR)，并 增加血管内皮生长因子的表达。此外，它们还表明，HUR本身 受低氧的调节。这项提案将重点放在Hur在 低氧对血管内皮细胞生长因子mRNA稳定性的调节建议的研究可能是 很重要，因为它们可能导致产生新的药物， 在转录后水平调控血管内皮生长因子的表达。具体的 该提案的目的如下：(1)确定罚款 HUR/VEGFm RNA复合体的结构。在这两个基因中产生突变 检测mRNA元件和HUR对血管内皮生长因子表达的影响； (2)设计HUR/VEGFm RNA复合体的选择性抑制剂；(3) 鉴定和克隆修饰其活性的Hur结合蛋白； 确定低氧如何调节Hur的表达；以及(5)确定 HUR如何稳定血管内皮生长因子基因的表达。

DESCRIPTION (Adapted from Investigator's Abstract): Vascular endothelial growth factor (VEGF) is an important mediator of angiogenesis in diabetic retinopathy, cancer and coronary heart disease. The expression of VEGF is tightly controlled to the cellular oxygen tension. Previous studies have shown that VEGF mRNA is markedly stabilized under hypoxic conditions. The molecular mechanisms underlying the regulation of VEGF mRNA stability are not well understood. In the preliminary results the applicant showed that the stability of VEGF mRNA is controlled by a cis acting element in its 3'UTR. They have also identified a trans acting factor (HuR) that binds to this element and increases the expression of VEGF. In addition, they show that HuR itself is regulated by hypoxia. This proposal will focus on the role of HuR in hypoxic regulation of VEGF mRNA stability. The proposed studies may be important since they could lead to the generation of new drugs that will regulate VEGF expression at the post transcriptional level. The specific aims of the proposal are the following: (1) To determine the fine structure of the HuR/VEGF mRNA complex. To generate mutations in both the mRNA element and HuR and determine their effect on VEGF expression; (2) To design selective inhibitors of HuR/VEGF mRNA complexes; (3) To identify and clone HuR associate proteins that modify its activity; (4) To determine how hypoxia regulates HuR expression; and (5) To determine how HuR stabilizes VEGF mRNA.