Role of KLK10 in Endothelial Biology and Atherosclerosis

KLK10 在内皮生物学和动脉粥样硬化中的作用

• 批准号: 10094078
• 负责人: Darian Williams
• 金额: $ 4.35万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-02-01 至 2022-01-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10094078
• 关键词: ARNT gene; Anti-Inflammatory Agents; Antiatherogenic; Antiinflammatory Effect; Area; Arterial Fatty Streak; Atherosclerosis; Biological Assay; Biology; Blood Vessels; Blood flow; Cause of Death; Cellular biology; Cholesterol; Coronary artery; Data; Development; Disease; Doctor of Philosophy; Endothelial Cells; Endothelium; Exercise; F2R gene; Functional disorder; G-Protein Signaling Pathway; GTP-Binding Proteins; Gene Proteins; Genes; Goals; Grant; Immunoprecipitation; In Vitro; Individual; Inflammation; Ischemic Stroke; Kininogenase; Laboratory Study; Lead; Lesion; Ligation; Light; Low Density Lipoprotein Receptor; MAPK1 gene; MAPK3 gene; Mechanics; Mediating; Mediator of activation protein; Medical; Mentors; Modeling; Molecular; Mus; Myocardial Infarction; PAR-1 Receptor; PI3K/AKT; Pathogenesis; Pathway interactions; Patients; Peptide Hydrolases; Peripheral arterial disease; Permeability; Pharmaceutical Preparations; Pharmacology; Prevention; Prevention therapy; Proteins; Publishing; Recombinants; Reporting; Role; Scientist; Serine Protease; Signal Pathway; Signal Transduction; Small Interfering RNA; Stents; Testing; Therapeutic; Thrombin; Training; Western Blotting; base; beta-arrestin; chronic inflammatory disease; endothelial dysfunction; hypercholesterolemia; in vivo; inhibitor/antagonist; insight; knock-down; mouse model; new therapeutic target; novel; novel therapeutics; overexpression; prevent; receptor; response; shear stress; success; targeted treatment; therapeutic protein

TITLE: Role of KLK10 in Endothelial Biology and Atherosclerosis PROJECT SUMMARY/ABSTRACT: Atherosclerosis is a chronic inflammatory disease of the arterial blood vessels that underlies the occurrence of heart attack, peripheral artery disease, and ischemic stroke; the leading causes of death worldwide. Currently cholesterol lowering statin drugs and stents are the most commonly used therapies for the prevention and treatment of atherosclerosis, however, despite their success, atherosclerosis is still the leading killer. Therefore, new therapeutics are needed to treat atherosclerosis. It is well known that atherosclerosis preferentially occurs in areas of disturbed blood flow while areas of stable flow are protected from developing atherosclerosis. Therefore, it is my goal to develop novel therapies to prevent and revert atherosclerotic diseases by studying the role of flow-dependent genes during my PhD training. In this grant, I will look to characterize one of the most flow-sensitive proteins, Kallikrein Related Peptidase 10 (KLK10), and its effects on atherosclerosis. KLK10 is a serine protease that has increased expression in endothelial cells under stable blood flow, while being dramatically reduced under disturbed blood flow conditions both in vivo and in vitro. KLK10 may also be important for regulating the development of atherosclerosis. We have preliminary data that KLK10 is able to lower endothelial cell inflammation and permeability, however the mechanism is unclear, and it is not known whether these individual effects lead to an overall anti-atherogenic effect. Based on preliminary data, I hypothesize that KLK10 inhibits endothelial inflammation, permeability, and atherosclerosis by mechanisms dependent on the Protease Activated Receptor 1 and 2 (PAR1 and PAR2) signaling pathway. In Aim 1, I will investigate the mechanisms by which KLK10 inhibits endothelial cell inflammation and permeability in a PAR1- and PAR2- dependent manner. This will involve in vivo and in vitro signaling assays using recombinant KLK10 (rKLK10) and pharmacological inhibitors or siRNAs of KLK10 and PAR1/2, under static and flow conditions. In Aim 2, I will test the ability of KLK10 to decrease atherosclerosis in vivo in a PAR1 and PAR2-dependent manner in vivo. In order to carry out these studies, I will use the Partial Carotid Ligation (PCL) mouse model developed in our lab, which allows us to induce disturbed blood flow and atherosclerosis in vivo. By injecting rKLK10 or KLK10 AAV into PCL mice, I will test whether KLK10 inhibits atherosclerosis progression, in both mice WT mice and mice without PAR1 or PAR2. These aims together will test whether KLK10 has a potential role as a therapeutic protein for the treatment of atherosclerosis. Furthermore, we will be able to delineate one of the key mechanisms underlying the flow-dependent development of atherosclerosis. Through this proposal, I will the obtain the necessary PhD training in mechanical biology and vascular pharmacology that will allow me to achieve my goals of developing and discovering novel therapies as a medical scientist.

标题：KLK10在内皮生物学和动脉粥样硬化中的作用 项目摘要/摘要： 动脉粥样硬化是动脉血管的慢性炎症性疾病 心脏病发作，周围动脉疾病和缺血性中风的发生；死亡的主要原因 全世界。目前，降低胆固醇的他汀类药物和支架是最常用的疗法 然而，预防和治疗动脉粥样硬化，尽管它们成功了，但动脉粥样硬化仍然是领先的 杀手。因此，需要新的治疗剂来治疗动脉粥样硬化。众所周知，动脉粥样硬化 优先发生在血流干扰的区域，而稳定流动的区域免受发展 动脉粥样硬化。因此，我的目标是开发新的疗法以预防和恢复动脉粥样硬化疾病 通过研究我的博士学位培训中流量依赖性基因的作用。在这笔赠款中，我将要描述一个 在流动敏感的蛋白中，Kallikrein相关的肽酶10（KLK10）及其对动脉粥样硬化的影响。 KLK10是一种丝氨酸蛋白酶，在稳定的血流下的内皮细胞中表达增加，而 在体内和体外的血流条件下，在干扰的血流条件下大大降低。 KLK10也可能是 对于调节动脉粥样硬化的发展至关重要。我们有KLK10能够的初步数据 下层内皮细胞​​炎症和渗透性，但是该机制尚不清楚，尚不清楚 这些个体效应是否导致总体抗动脉粥样硬化作用。基于初步数据，我 假设KLK10通过机制抑制内皮炎症，渗透性和动脉粥样硬化 取决于蛋白酶激活的受体1和2（PAR1和PAR2）信号通路。在AIM 1中，我会 研究KLK10抑制par1-中内皮细胞炎症和渗透性的机制 和par2依赖的方式。这将涉及使用重组KLK10的体内和体外信号传导测定 （RKLK10）在静态和流动条件下，KLK10和PAR1/2的药理抑制剂或siRNA。在 AIM 2，我将测试KLK10以PAR1和PAR2依赖性方式减少体内动脉粥样硬化的能力 体内。为了进行这些研究，我将使用开发的部分颈动脉连接（PCL）模型 在我们的实验室中，这使我们能够在体内诱导血液流动和动脉粥样硬化。通过注射RKLK10或 KLK10 AAV进入PCL小鼠，我将测试KLK10是否抑制了两只小鼠WT小鼠的动脉粥样硬化进展 和没有PAR1或PAR2的小鼠。这些目标共同测试KLK10是否具有潜在的作用 治疗动脉粥样硬化的治疗蛋白。此外，我们将能够描述其中之一 动脉粥样硬化流动发展的基础机制。通过这个建议，我将 获得机械生物学和血管药理学中必要的博士学位培训，这将使我能够实现 我作为医学科学家开发和发现新颖疗法的目标。