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.

KLK10在内皮生物学和动脉粥样硬化中的作用 项目摘要/摘要： 动脉粥样硬化是一种慢性炎症性动脉血管疾病，它是 心脏病发作、外周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，降胆固醇他汀类药物和支架是最常用的治疗方法 动脉粥样硬化的防治，尽管取得了成功，但动脉粥样硬化仍然是主要的 杀手。因此，需要新的治疗方法来治疗动脉粥样硬化。众所周知，动脉粥样硬化 优先发生在血流紊乱的区域，而血流稳定的区域则受到保护，不会发展 动脉硬化。因此，开发新的疗法来预防和逆转动脉粥样硬化性疾病是我的目标。 通过研究流动依赖基因在我的博士培训中的作用。在这笔赠款中，我将着眼于描述一个 最敏感的流动蛋白，激肽释放酶相关的多肽酶10(KLK10)及其在动脉粥样硬化中的作用。 KLK10是一种丝氨酸蛋白酶，在血流稳定的情况下，内皮细胞表达增加，而 在体内和体外血液流动受扰的情况下显著降低。KLK10也可能是 对于调节动脉粥样硬化的发展具有重要意义。我们有初步数据表明KLK10能够 降低内皮细胞炎症和通透性，但其机制尚不清楚，也不清楚。 这些单独的影响是否会导致整体的抗动脉粥样硬化效应。根据初步数据，我 假设KLK10通过机制抑制内皮细胞炎症、通透性和动脉粥样硬化 依赖于蛋白水解酶激活受体1和2(PAR1和PAR2)信号通路。在《目标1》中，我将 探讨KLK10抑制血管内皮细胞炎症和通透性的机制 和PAR2依赖的方式。这将涉及使用重组KLK10进行体内和体外信号分析 (RKLK10)以及KLK10和PAR1/2的药理抑制剂或siRNA，在静态和流动条件下。在……里面 目的2，以PAR1和PAR2依赖的方式检测KLK10在体内减轻动脉粥样硬化的能力 在活体内。为了进行这些研究，我将使用开发的部分颈动脉结扎(PCL)小鼠模型 在我们的实验室里，这使得我们可以在体内诱导血液流动障碍和动脉粥样硬化。通过注射rKLK10或 将KLK10 AAV导入PCL小鼠，我将检测KLK10是否抑制动脉粥样硬化的进展，在两只小鼠中均为WT小鼠 和没有PAR1或PAR2的小鼠。这些目标加在一起将考验KLK10是否具有潜在的 治疗动脉粥样硬化的治疗蛋白。此外，我们将能够描绘出其中一个关键 动脉粥样硬化血流依赖性发展的潜在机制。通过这项提议，我将 获得必要的机械生物学和血管药理学博士培训，这将使我能够实现 作为一名医学科学家，我的目标是开发和发现新的疗法。