Role of KLK10 in Endothelial Biology and Atherosclerosis

标题：KLK 10在内皮生物学和动脉粥样硬化中的作用 项目总结/摘要： 动脉粥样硬化是动脉血管的慢性炎症性疾病，其是动脉粥样硬化的基础。 心脏病发作、外周动脉疾病和缺血性卒中的发生;死亡的主要原因 国际吧目前，降低胆固醇的他汀类药物和支架是最常用的治疗方法。 预防和治疗动脉粥样硬化，然而，尽管他们的成功，动脉粥样硬化仍然是主要的 杀手因此，需要新的疗法来治疗动脉粥样硬化。众所周知，动脉粥样硬化 优先发生在血流受干扰的区域，而血流稳定的区域被保护免于发展 动脉粥样硬化因此，我的目标是开发新的治疗方法来预防和逆转动脉粥样硬化疾病 通过在我的博士培训期间研究依赖于血流的基因的作用。在这次资助中，我将描述一个 最流动敏感的蛋白质，激肽释放酶相关肽酶10（KLK 10），及其对动脉粥样硬化的影响。 KLK 10是一种丝氨酸蛋白酶，在稳定血流条件下，其在内皮细胞中的表达增加， 在体内和体外受干扰的血流条件下显著降低。KLK 10也可能是 对调节动脉粥样硬化的发展很重要。我们有初步数据表明，KLK 10能够 降低内皮细胞炎症和通透性，但机制尚不清楚，也不清楚 这些个体效应是否导致整体抗动脉粥样硬化效应。根据初步数据，我 假设KLK 10通过以下机制抑制内皮炎症、渗透性和动脉粥样硬化 依赖于蛋白酶激活受体1和2（PAR 1和PAR 2）信号通路。在目标1中，我将 研究KLK 10抑制内皮细胞炎症和PAR 1 - 1通透性的机制。 和PAR 2依赖性方式。这将涉及使用重组KLK 10的体内和体外信号传导测定 （rKLK 10）和KLK 10和PAR 1/2的药理学抑制剂或siRNA。在 目的2，我将测试KLK 10在体内以PAR 1和PAR 2依赖性方式减少动脉粥样硬化的能力 in vivo.为了进行这些研究，我将使用部分颈动脉结扎（PCL）小鼠模型， 在我们的实验室中，这使我们能够在体内诱导血流紊乱和动脉粥样硬化。通过注射rKLK 10或 将KLK 10 AAV导入PCL小鼠，我将在两种小鼠WT小鼠中测试KLK 10是否抑制动脉粥样硬化进展。 和没有PAR 1或PAR 2的小鼠。这些目标将共同测试KLK 10是否具有潜在的作用， 用于治疗动脉粥样硬化的治疗性蛋白质。此外，我们将能够描绘出一个关键的 动脉粥样硬化的血流依赖性发展的潜在机制。通过这一建议，我将 获得必要的机械生物学和血管药理学博士培训，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。

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.