Biologic Role of Cytomegalovirus in Endothelial Cell Inflammation and Atheroscler

说明(申请人提供)：这项提案的意义在于，它侧重于心血管疾病，这是工业化国家死亡的主要原因。动脉粥样硬化主要发生在动脉树有分支和弯曲的区域，这些区域的血液流动受到干扰，切应力低且不均匀。越来越多的证据表明，具有高切应力的层流血流调节内皮细胞(ECs)的基因表达，以保护血管内皮细胞免受动脉粥样硬化、炎症和凝血的影响，并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，巨细胞病毒在疾病过程中的作用机制是什么？许多研究表明，HCMV感染可诱导内皮细胞、血管内皮细胞和单核/巨噬细胞表达致动脉粥样硬化基因，但这些研究都是在无流动或剪应力的静态细胞培养中进行的。Deborah Spector实验室是第一个研究暴露在不同流动和剪应力条件下的主动脉内皮细胞感染HCMV的实验室。我们假设，血流条件影响HCMV与内皮细胞的相互作用，进而调节内皮细胞的功能以及与白细胞和平滑肌细胞的相互作用，从而导致病变的形成。为了解决有关HCMV感染在EC炎症中的问题，需要详细了解HCMV的发病机制和体内动物模型。这项建议的新奇之处在于，它通过跨学科的方法解决了人巨细胞病毒感染和血流动力学在动脉粥样硬化中的作用。它汇集了 Deborah Spector实验室在HCMV和MCMV的分子和细胞生物学方面拥有丰富的经验，Stephen Spector实验室在HCMV发病机制和转化医学方面拥有丰富的经验，Joseph Witztom在动脉粥样硬化的体内发病机制方面拥有丰富的知识和技术专长，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，我们将确定高剪应力和低剪应力(HSS和LSS)下HCMV和ECs之间的双向相互作用。目的2研究人巨细胞病毒感染内皮细胞后，在高糖和低糖条件下对免疫刺激和NA�vE PBMC的黏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。这项建议的长期目标是为动脉粥样硬化的发病机制提供新的见解。实现这一目标将有助于制定旨在预防和治疗动脉粥样硬化性疾病的新战略。

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.