Mechanism of chymase activation in endothelial cells

内皮细胞食糜酶激活机制

• 批准号: 8636206
• 负责人: YUPING WANG
• 金额: $ 18万
• 依托单位: LOUISIANA STATE UNIV HSC SHREVEPORT
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-02-01 至 2016-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8636206
• 关键词: Acetylation; Angiotensin II; Applications Grants; Biological Models; Blood Vessels; Cardiovascular Diseases; Cardiovascular system; Chymase; Diabetes Mellitus; Endothelial Cells; Endothelium; Enzymes; Epigenetic Process; Functional disorder; Gene Transfer; Generations; Genes; Histone Acetylation; Histone Deacetylase; Histone Deacetylase Inhibitor; Histones; Hypertension; Inflammatory; Lead; Link; Mediating; Mediator of activation protein; Metabolic Diseases; Molecular; Muscle Contraction; Pathogenesis; Peptide Hydrolases; Peptidyl-Dipeptidase A; Placenta; Play; Pre-Eclampsia; Pregnancy; Proteins; Regulation; Research Design; Role; Serine Protease; Signal Transduction; Small Interfering RNA; Techniques; Testing; Time; Tissues; Up-Regulation; Vascular Diseases; Vascular Endothelial Cell; Vascular Endothelial Growth Factor Receptor; Vasoconstrictor Agents; Woman; base; chymotrypsin; endothelial dysfunction; gain of function; histone acetyltransferase; histone modification; human HDAC4 protein; in vitro Model; inhibitor/antagonist; insight; loss of function; normotensive; novel; novel therapeutics; pregnant; public health relevance; response; vasoconstriction

DESCRIPTION (provided by applicant): The objective of this study is to explore the mechanism of increased chymase expression in endothelial cells (ECs) and to test our hypothesis that histone modification leads to chymase activation in vascular ECs in preeclampsia (PE). Chymase, a chymotrypsin-like serine protease (CLP), is a significant contributor to cardiovascular diseases, including hypertension and diabetes. Chymase is an ACE-independent angiotensin II (Ang II) converting enzyme. Ang II, a potent vasoconstrictor, plays a key role in increased vasoconstriction in PE. Chymase expression is increased in maternal vessel endothelium in women with PE compared to normotensive controls. CLP/chymase derived from PE placenta is not only responsible for triggering EC chymase activation, but also contributes to placental sFlt-1 release in PE. However, the mechanism that underlies chymase activation remains elusive. To explore the mechanism of EC chymase activation in PE, in our preliminary study that using an in vitro model system that mimics the increased EC chymase expression in PE, we found that specific HDAC inhibition directly contributes to chymase regulation. In this study, we propose for the first time to explore the epigenetic basis of placenta-mediated activation of chymase in the endothelium, specifically, we will test the hypothesis that aberrant epigenetic regulation (histone modification) mediated endothelial chymase activation in PE. This hypothesis will be tested in 2 specific aims. Aim 1 will demonstrate how placental factor-mediated histone modification leads to endothelial chymase expression in PE and Aim 2 will demonstrate how histone acetylation leads to chymase activation in ECs. Using state-of-the-art techniques, chymase gene transfer approach, siRNA and specific inhibitors and blockers of target molecules, we will define how histone modification mediates chymase expression in EC in PE. These studies represent the first attempt to investigate the role of epigenetic regulation, which connects placenta dysfunction, chymase activation, and EC phenotypic changes that occur in PE. Results obtained from this study will lead to novel mechanistic explanation of chymase activation and its pathological effects on endothelial function in PE and will provide rationale for developing a new therapeutic strategy to treat chymase activation associated vascular diseases and metabolic disorders.

描述（由申请人提供）：本研究的目的是探索内皮细胞（EC）中糜酶表达增加的机制，并验证我们的假设，即组蛋白修饰导致先兆子痫（PE）血管EC中糜酶激活。糜蛋白酶是一种糜蛋白酶样丝氨酸蛋白酶（CLP），是心血管疾病（包括高血压和糖尿病）的重要贡献者。糜酶是一种ACE非依赖性血管紧张素II（Ang II）转化酶。血管紧张素II是一种强有力的血管收缩剂，在PE中血管收缩增加中起关键作用。与血压正常的对照组相比，PE妇女的母血管内皮中糜蛋白酶表达增加。来自PE胎盘的CLP/糜酶不仅负责触发EC糜酶激活，而且有助于PE中胎盘sFlt-1的释放。然而，糜酶激活的机制仍然难以捉摸。为了探索PE中EC糜酶激活的机制，在我们的初步研究中，使用模拟PE中EC糜酶表达增加的体外模型系统，我们发现特异性HDAC抑制直接有助于糜酶调节。在这项研究中，我们首次提出探索胎盘介导的内皮细胞糜酶激活的表观遗传基础，具体来说，我们将测试的假设，即异常表观遗传调节（组蛋白修饰）介导的内皮细胞糜酶激活PE。这一假设将在2个具体目标中进行检验。目标1将 Aim 2将证明胎盘因子介导的组蛋白修饰如何导致PE中内皮糜酶表达，Aim 2将证明组蛋白乙酰化如何导致EC中糜酶激活。利用最先进的技术，糜酶基因转移的方法，siRNA和特异性抑制剂和靶分子的阻断剂，我们将确定如何组蛋白修饰介导的糜酶表达在EC在PE。这些研究代表了第一次尝试调查表观遗传调控的作用，它连接胎盘功能障碍，糜酶激活和EC表型变化，发生在PE。从这项研究中获得的结果将导致新的机制解释糜酶激活及其对PE内皮功能的病理影响，并将为开发一种新的治疗策略来治疗糜酶激活相关的血管疾病和代谢紊乱提供理论基础。