Biologic Role of Cytomegalovirus in Endothelial Cell Inflammation and Atheroscler

描述（由申请人提供）：该提案的重要性在于它重点关注心血管疾病，心血管疾病是工业化国家死亡的主要原因。动脉粥样硬化优先发生在具有分支和弯曲的动脉树区域，其中血流受到干扰并且剪切应力低且不均匀。越来越多的证据表明，高剪切应力的层流血流可调节内皮细胞 (EC) 的基因表达，以防止动脉粥样硬化、炎症和凝血，并且受干扰的血流会上调促动脉粥样硬化、促炎和促凝血基因。长期以来，人们一直怀疑人类巨细胞病毒（HCMV）感染是动脉粥样硬化和血管成形术后再狭窄等血管疾病的危险因素。关键问题是HCMV在疾病过程中发挥作用的机制是什么？许多研究表明，HCMV 感染会诱导内皮细胞、平滑肌细胞和单核细胞/巨噬细胞中促动脉粥样硬化基因表达，但所有这些研究都是在静态细胞培养中进行的，其中没有流动或剪切应力。 Deborah Spector 实验室是第一个研究暴露于不同流动和剪切应力条件下的主动脉 EC 的 HCMV 感染的实验室。我们假设流动条件影响 HCMV 与 EC 的相互作用，这反过来又调节 EC 的功能以及与白细胞和平滑肌细胞的相互作用，从而导致病变形成。为了解决 EC 炎症中 HCMV 感染的问题，需要详细了解 HCMV 发病机制以及体内动物模型。该提案的新颖之处在于它通过跨学科的方法解决了 HCMV 感染和血流动力学在动脉粥样硬化中的作用。它汇集了各领域广泛的专业知识 Deborah Spector 实验室在 HCMV 和 MCMV 的分子和细胞生物学方面的研究、Stephen Spector 实验室在 HCMV 发病机制和转化医学方面的丰富经验以及 Joseph Witztum 在动脉粥样硬化体内发病机制方面的丰富知识和技术专长，以检验我们的假设并评估 HCMV 在动脉粥样硬化中的潜在作用。提出了三个具体目标。在目标 1 中，我们将确定 HCMV 和 EC 在高剪切应力与低剪切应力（HSS 与 LSS）下的双向相互作用。在目标 2 中，我们将确定在 HSS 和 LSS 条件下，EC 的 HCMV 感染对免疫引发和初始 PBMC 的粘附和跨内皮迁移的影响。在目标 3 中，我们将利用体内研究来确定 MCMV 对 ApoE-/- 小鼠动脉粥样硬化模型的影响。该提案的长期目标是为动脉粥样硬化的发病机制提供新的见解。这一目标的实现将促进旨在预防和治疗动脉粥样硬化疾病的新策略的开发。

DESCRIPTION (provided by applicant): The significance of this proposal is that it focuses on cardiovascular diseases which represent a leading cause of mortality in industrialized nations. Atherosclerosis preferentially develops in regions of the arterial tree with branches and curvatures where blood flow is disturbed and shear stress is low and non- uniform. There is increasing evidence that laminar blood flow with high shear stress modulates gene expression in endothelial cells (ECs) to protect against atherosclerosis, inflammation and coagulation, and that disturbed flow upregulates proatherosclerotic, proinflammatory, and procoagulant genes. It has long been suspected that human cytomegalovirus (HCMV) infection is a risk factor for vascular disease such as atherosclerosis and restenosis following angioplasty. The key question is what is the mechanism underlying HCMV's role in the disease process? Many studies have shown that HCMV infection induces proatherogenic gene expression in ECs, smooth muscle cells and monocytes/macrophages, but all these studies were performed in static cell culture, where there is no flow or shear stress. The Deborah Spector lab is the first t study HCMV infection of aortic ECs exposed to varying conditions of flow and shear stress. We hypothesize that flow conditions affect HCMV interaction with ECs and that this in turn modulates the EC functions and interactions with leukocytes, and smooth muscle cells to lead to lesion formation. Detailed knowledge of HCMV pathogenesis as well as in vivo animal models are required in order to address questions regarding the HCMV infection in EC inflammation. The novelty of this proposal is that it addresses the roles of HCMV infection and flow dyamics in atherosclerosis by an interdisciplinary approach. It brings together the extensive expertise in the Deborah Spector lab on molecular and cellular biology of HCMV and MCMV, the broad experience in the Stephen Spector lab on HCMV pathogenesis and translational medicine, and the vast knowledge and technical expertise of Joseph Witztum on the in vivo pathogenesis of atherosclerosis to test our hypothesis and assess the potential role of HCMV in atherosclerosis. Three Specific Aims are proposed. In Aim 1, we will determine the bi-directional interactions between HCMV and ECs under high vs. low shear stress (HSS vs. LSS). In Aim 2, we will determine the effect of HCMV infection of ECs on adhesion and transendothelial migration of Immunologically primed and na�ve PBMCs under conditions of HSS and LSS. In Aim 3, we will utilize in vivo studies to define the impact of MCMV on the ApoE-/- mouse model of atherosclerosis. The long- term objective of this proposal is to provide novel insights into the pathogenesis of atherosclerosis. Accomplishment of this goal will facilitate the development of new strategies designed to prevent and treat atherosclerotic disease.