microRNA regulation of endothelial functions

microRNA对内皮功能的调节

基本信息

批准号：
8444656
负责人：
William C Sessa
金额：
$ 44.77万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-04-06 至 2014-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8444656
关键词：
3&apos Untranslated Regions Area Base Pairing Blood Vessels Blood flow Cardiovascular Diseases Cardiovascular system Cell Cycle Cellular biology Data Dicer Enzyme Employee Strikes Endothelial Cells Endothelium Enzymes Exhibits Fibroblast Growth Factor 2 Functional disorder Gene Expression Genome Grant Growth Factor Heart Diseases Human In Vitro Ischemia Messenger RNA MicroRNAs Modeling Morphogenesis Mus Pathway interactions Pattern Pharmaceutical Preparations Phenotype Physiological Process Proteins Public Health Quality of life Regulation Regulator Genes Research Research Support Role Signal Transduction Testing Time Tissues Transduction Gene Transgenic Mice Translations Untranslated RNA Vascular Endothelial Growth Factors Work angiogenesis atherogenesis cell growth cell type human DICER1 protein human NOS3 protein implantation improved in vivo insight mimicry novel therapeutic intervention public health relevance research study response stem therapeutic target tumor

项目摘要

DESCRIPTION (provided by applicant): The specific roles of miRNAs in vascular function are just beginning to be explored. Our previous work has shown that reduction of overall miRNA levels via Dicer silencing in human endothelial cells (EC) strongly regulates angiogenic gene expression and impairs aspects of in vitro angiogenesis including EC growth and morphogenesis. These in vitro experiments are supported by new exciting preliminary data in vivo data using mice conditionally lacking the rate limiting enzyme (Dicer) in miRNA synthesis in EC. Mice lacking Dicer in EC (EC specific Dicer KO) are viable but exhibit impaired post-natal angiogenic responses. In order to dissect the relationships between signal transduction, gene expression and miRNAs in EC, we show that treatment of EC with VEGF stimulates time-dependent changes in global miRNA profiles and show that components of the miRNA cluster, miR 17-92 can regulate aspects of VEGF induced cell growth and morphogenesis. In addition, we have identified a specific miRNA (miR-155) that regulates the levels of endothelial nitric oxide synthase (eNOS) in cultured EC. Thus, we hypothesize that Dicer generated endothelial miRNAs regulate the extent of angiogenesis and blood flow control in vivo by regulating mRNA levels and/or the translational stability of important proteins. We will: 1. Identify and characterize angiogenic growth factor regulated miRNAs in EC; 2. Define the roles of miR 17-92 in models of angiogenesis and 3. Define the roles of miR-155 in regulating eNOS function in vitro and in vivo. Collectively, this work will facilitate the understanding of the importance of miRNAs in EC biology and shed insights into their role as potential therapeutics targets.

描述（由申请人提供）：miRNA 在血管功能中的具体作用刚刚开始被探索。我们之前的工作表明，通过人内皮细胞 (EC) 中 Dicer 沉默降低总体 miRNA 水平，会强烈调节血管生成基因表达，并损害体外血管生成的各个方面，包括 EC 生长和形态发生。这些体外实验得到了新的令人兴奋的初步数据的支持，体内数据使用条件性缺乏 EC 中 miRNA 合成限速酶 (Dicer) 的小鼠。 EC 中缺乏 Dicer 的小鼠（EC 特异性 Dicer KO）可以存活，但表现出产后血管生成反应受损。为了剖析 EC 中信号转导、基因表达和 miRNA 之间的关系，我们发现用 VEGF 治疗 EC 会刺激整体 miRNA 谱的时间依赖性变化，并表明 miRNA 簇的组成部分 miR 17-92 可以调节各个方面VEGF 诱导细胞生长和形态发生。此外，我们还发现了一种特定的 miRNA (miR-155)，它可以调节培养的 EC 中内皮一氧化氮合酶 (eNOS) 的水平。因此，我们假设 Dicer 产生的内皮 miRNA 通过调节 mRNA 水平和/或重要蛋白质的翻译稳定性来调节体内血管生成和血流控制的程度。我们将： 1. 鉴定并表征 EC 中血管生成生长因子调节的 miRNA； 2. 定义 miR 17-92 在血管生成模型中的作用，以及 3. 定义 miR-155 在体外和体内调节 eNOS 功能中的作用。总的来说，这项工作将有助于理解 miRNA 在 EC 生物学中的重要性，并深入了解它们作为潜在治疗靶点的作用。