Vessel Shear Stress & Cytomegalovirus Disease: Molecular Basis of Atherosclerosis

描述（由申请人提供）：在发达国家，心脏病和中风是导致死亡和长期残疾的主要原因。心血管疾病的主要潜在原因是动脉粥样硬化，它被广泛认为是由内皮细胞损伤或功能障碍引发的炎症性疾病。人类巨细胞病毒（HCMV）是一种广泛传播的人类病原体，长期以来被认为是心血管疾病的危险因素。动脉粥样硬化的发病机制涉及多种细胞类型之间的相互作用，通常发生在大到中等动脉的分支或弯曲区域，这些区域暴露于受干扰的血流模式和低流体剪切应力。迄今为止，HCMV感染在动脉粥样硬化中的作用仅在静态条件下进行了探索。鉴于细胞间相互作用的重要性和血流在动脉粥样硬化中的作用，本研究计划的主要目标是利用血流模型系统在生理背景下揭示HCMV诱导动脉粥样硬化的关键机制。我们计划探索HCMV和内皮细胞在均匀和湍流高或低剪切应力条件下的双向相互作用。为了做到这一点，我们将在室内培养内皮细胞，这将允许介质在模拟血液流动的细胞上进行调节流动。均匀流室和阶梯流室的使用将使我们能够模拟不同类型的流动模式和发生在脉管系统不同部分的剪切应力。利用这些系统，我们计划分析暴露于不同流动模式和剪切应力下的内皮细胞中HCMV感染的影响，因为它与病毒进展和蛋白和RNA水平上的促动脉粥样硬化基因表达有关。此外，我们计划确定在血流背景下内皮细胞的HCMV感染最终如何调节外周血白细胞的粘附和跨内皮迁移，这是动脉粥样硬化病变发展的主要初始步骤。总体而言，本研究计划旨在探索HCMV与内皮细胞和白细胞在血流生理背景下的相互作用，以阐明HCMV诱导动脉粥样硬化的机制。考虑到HCMV在人群中的患病率（60-80%的成年人）、与动脉粥样硬化相关的死亡率及其对医疗保健费用的影响，有必要确定HCMV感染在人类动脉发生的生理条件下动脉粥样硬化进展中的作用。该研究计划的长期目标是为动脉粥样硬化的病毒发病机制提供新的见解，从而为预防与HCMV感染相关的内皮细胞损伤提供具有成本效益的新策略。

DESCRIPTION (provided by applicant): Heart disease and stroke are the principal causes of mortality and long-term disability in developed nations. The major underlying cause of cardiovascular diseases is atherosclerosis, which is widely recognized as an inflammatory disease initiated by endothelial cell injury or dysfunction. Human cytomegalovirus (HCMV), a widely disseminated human pathogen, has long been proposed as a risk factor for cardiovascular disease. The pathogenesis of atherosclerosis involves interactions between multiple cell types and typically occurs in large to medium-sized arteries in regions that are branched or curved, which are exposed to disturbed patterns of blood flow and low fluid shear stress. To date, the role of HCMV infection in atherosclerosis has only been explored in static conditions. Given the importance cell-cell interactions and the role of blood flow in atherogenesis, the major goal of this research plan is to uncover key mechanisms in HCMV induced atherosclerosis in a physiological context using a flow model system. We plan to explore the bi-directional interactions of HCMV and endothelial cells under conditions of high or low shear stress in uniform and turbulent flow. To do this we will culture endothelial cells in chambers that will allow for the regulated flow of media over cells simulating blood flow. The use of uniform and step-flow chambers will allow us to simulate different types of flow patterns and shear stresses that occur in different parts of the vasculature. Using these systems we plan to analyze the effects of HCMV infection in endothelial cells exposed to varying flow patterns and shear stress as it relates to viral progression and proatherosclerotic gene expression at the protein and RNA level. In addition, we plan to determine how HCMV infection of endothelial cells in the context of flow ultimately modulates the adhesion and transendothelial migration of peripheral blood leukocytes, which is the major initial step in the development of atherosclerotic lesions. As a whole, this research plan is intended to explore the interactions between HCMV, endothelial cells, and leukocytes in the physiological context of blood flow in order to elucidate the mechanisms of HCMV induced atherosclerosis. Given the prevalence of HCMV in the human population (60-80% of adults), the mortality associated with atherosclerosis, and its impact on healthcare costs, there is a great need to determine the role of HCMV infection in the progression of atherogenesis in the physiological conditions that occur in the human artery. The long-term goal of this research plan is to provide novel insights into viral pathogenesis of atherosclerosis leading to cost effective new strategies for prevention of endothelial cell damage associated with HCMV infection. PUBLIC HEALTH RELEVANCE: Atherosclerosis is the major cause cardiovascular diseases (the leading cause of mortality in developed countries) and its impact on health care costs is enormous. Hemodynamic factors are major regulators of atherosclerosis, and human cytomegalovirus (HCMV) infection has been proposed as an important factor in disease progression. The proposed project employs novel approaches to study the effects of HCMV infection on vascular functions in a dynamic flow environment, and the results will fill the current gap in our understanding of bi-directional interactions between virus and vascular cells in physiological or pathological environments, thus facilitating the discovery of new therapies.