DETERMINING THE INFLUENCES OF HCMV INFECTION IN VASCULAR SCLEROSIS

确定 HCMV 感染对血管硬化的影响

• 批准号: 7721404
• 负责人: JAY A NELSON
• 金额: $ 2.4万
• 依托单位: BATTELLE PACIFIC NORTHWEST LABORATORIES
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-08 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7721404
• 关键词: Acceleration; Accounting; Allografting; Angiogenic Factor; Antiviral Agents; Antiviral Therapy; Atherosclerosis; Blood Vessels; Cell Proliferation; Cell secretion; Cells; Chronic; Complex; Computer Retrieval of Information on Scientific Projects Database; Cytomegalovirus; Cytomegalovirus Infections; DNA Microarray Chip; DNA Microarray format; Development; Disease; Endothelial Cells; Event; Fibroblasts; Foscarnet; Funding; Ganciclovir; Genes; Goals; Grant; Growth Factor; Heart; Heart Transplantation; Human; Immune; Inflammation; Injury; Institution; Laboratories; Mass Spectrum Analysis; Mechanics; Mediating; Microarray Analysis; Modeling; Process; Production; Proteins; Rattus; Research; Research Personnel; Resources; Role; Sclerosis; Smooth Muscle Myocytes; Source; Transplant Recipients; Transplantation; United States National Institutes of Health; Vascular Diseases; Viral; Virus; Wound Healing; angiogenesis; base; cytokine; heart allograft; in vitro Assay; intracellular protein transport; macrophage; migration; monocyte; mutant; protein transport; restenosis; tool; trafficking

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The long-term goal of this project is to determine the role of human cytomegalovirus (HCMV) in the acceleration of vascular diseases such as atherosclerosis, restenosis, and transplant vascular sclerosis (TVS). All of these diseases are the result of either mechanical or immune-mediated injury followed by inflammation and subsequent smooth muscle cell (SMC) proliferation and migration from the vessel media to the intima that culminates in vessel narrowing. Besides SMC, other key cells in the development of vascular disease are monocyte-derived macrophages (MDM) and endothelial cells (EC) that also serve as cellular reservoirs of HCMV. Studies by our group have demonstrated that rat CMV (RCMV) significantly accelerates TVS and chronic rejection (CR) in heart allografts in the rat transplant model. In addition, treatment of recipient rats with antiviral drug ganciclovir results in prolonged survival of the allograft indication the CMV replication is a requirement for acceleration of disease. Similar observations have been made in HCMV infected human heart transplant patients receiving antiviral therapy. However, although virus persists in the allograft throughout the course of disease, the amount of allograft cells directly infected with RCMV does not account for the global effects that the virus has on the acceleration of TVS and CR. Recently, an analysis of up- and down-regulated cellular genes by DNA microarray analysis in infected allografts in comparison to native heart has demonstrated that RCMV up-regulates genes involved in wound healing (WH) and angiogenesis (AG). Consistent with this result, supernatants from HCMV infected cells (HCMV secretome) were shown in in vitro assays to induce WH and AG. Recent results from our laboratory indicate that HCMV induction of the WH/AG factors requires viral replication since neither UV inactivated virus nor the antiviral drug foscarnet treated infected cells are capable of production of these secreted factors. These latter observations correlate with antiviral treatment of CMV infected transplant recipients. In addition, MS analysis of these HCMV supernatants identified over 1000 virus-induced cellular proteins, some of which are involved in angiogenesis and wound healing events. We hypothesize that one mechanism for HCMV acceleration of TVS is mediated through viral induction of cellular secretion of cytokines and growth factors from virus-infected EC and macrophages that promote WH and AG in the allograft, resulting in the acceleration of TVS and CR. The cytokines and growth factors in cellular secretomes involved in WH and AG is complex. Using wild-type (WT) and mutant viruses and experimental approaches that utilize sensitive mass spectrometry-based analyses as tools to dissect these events will allow us to analyze the cellular secretomes of mutant HCMV-infected cells that do not promote WH and AG; to characterize the synthesis, trafficking, and localization of the secretome proteins; and to identify combinations of cytokines and growth factors that are important in these processes. Therefore, to prove this hypothesis and identify the HCMV genes that induce the secretion of these cytokines and growth factors we propose the following: Specific Aim 1: What is the spectrum of cytokines and growth factors in the HCMV secretome that induce WH and AG? Specific Aim 2: Are there cell or HCMV strain specific differences in the secretomes that indue WH and AG? Specific Aim 3: Are there differences in the secretomes of HCMV infected fibroblasts, macrophages, EC and SMC? How does HCMV infection modify protein trafficking in host cells?

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 该项目的长期目标是确定人类巨细胞病毒（HCMV）在诸如动脉粥样硬化，再狭窄和移植血管硬化症（TVS）等血管疾病加速中的作用。 所有这些疾病都是机械或免疫介导的损伤的结果，随后是炎症和随后的平滑肌细胞（SMC）增殖，以及从血管培养基到最终导致血管变窄的内膜的迁移。 除SMC外，血管疾病发展中的其他关键细胞是单核细胞衍生的巨噬细胞（MDM）和内皮细胞（EC），它们也用作HCMV的细胞储层。 我们小组的研究表明，大鼠移植模型中，大鼠CMV（RCMV）在心脏同种异体移植物中显着加速了电视和慢性排斥（CR）。 此外，用抗病毒药物ganciclovir治疗接受者大鼠的治疗导致同种异体移植指示的长期存活率表明CMV复制是疾病加速的必要条件。 在接受抗病毒疗法的HCMV感染的人心脏移植患者中，已经进行了类似的观察。 然而，尽管在整个疾病过程中的同种异体移植中持续存在病毒，但直接感染RCMV的同种异体移植细胞的量并不能说明病毒对TVS和CR加速的全球影响。最近，与天然心脏相比，通过DNA微阵列分析对上调和下调的细胞基因的分析表明，RCMV上调了与伤口愈合（WH）和血管生成（AG）的基因。 与该结果一致，在体外测定中显示了来自HCMV感染细胞（HCMV分泌组）的上清液，以诱导WH和AG。 我们实验室的最新结果表明，HCMV诱导WH/AG因子需要病毒复制，因为紫外线灭活病毒均不灭活病毒，也不是抗病毒药FOSCARNET治疗的受感染细胞能够产生这些分泌的因素。 这些后一种观察结果与CMV感染的移植受者的抗病毒治疗相关。 此外，对这些HCMV上清液的MS分析确定了1000多个病毒诱导的细胞蛋白，其中一些涉及血管生成和伤口愈合事件。 我们假设电视的HCMV加速度的一种机制是通过病毒诱导细胞因子的细胞分泌以及来自病毒感染的EC和巨噬细胞的生长因子的病毒诱导，从而促进同种异体移植物中WH和AG的生长因子，从而导致电视和CR的加速。 WH和Ag涉及的细胞分泌组中的细胞因子和生长因子很复杂。 使用野生型（WT）和突变病毒和实验方法，这些方法利用基于敏感的质谱分析作为解剖这些事件的工具，将使我们能够分析未促进WH和AG的突变HCMV感染的细胞的细胞秘密；表征分泌蛋白的合成，贩运和定位；并确定在这些过程中重要的细胞因子和生长因子的组合。 因此，为了证明这一假设并确定诱导这些细胞因子和生长因子分泌的HCMV基因，我们提出了以下内容： 具体目标1：诱导WH和Ag的HCMV分泌组中细胞因子和生长因子的谱是什么？ 特定目标2：在诱发WH和Ag的秘密组中是否存在细胞或HCMV菌株特异性差异？ 特定目的3：HCMV感染的成纤维细胞，巨噬细胞，EC和SMC的秘密构成是否存在差异？ HCMV感染如何改变宿主细胞中的蛋白质运输？