The role of CMV secretome in the acceleration of transplant vascular sclerosis

CMV分泌组在加速移植血管硬化中的作用

• 批准号: 8213656
• 负责人: JAY A NELSON
• 金额: $ 63.63万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-01 至 2013-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8213656
• 关键词: Acceleration; Accounting; Affect; Agonist; Allografting; Angiogenic Factor; Antiviral Agents; Antiviral Therapy; Atherosclerosis; Biological Assay; Blood Vessels; Cell Proliferation; Cell secretion; Cells; Chronic; Complex; Cytomegalovirus; DNA Microarray Chip; Development; Disease; Endothelial Cells; Event; Fibroblasts; Foscarnet; Ganciclovir; Gel; Gene Expression Microarray Analysis; Genes; Goals; Growth Factor; Heart; Heart Transplantation; Human; Immune; In Vitro; Infection; Inflammation; Injury; Knock-out; Laboratories; Mass Spectrum Analysis; Mechanics; Mediating; Microarray Analysis; Modeling; Mutation; Organ; Organ Transplantation; Process; Production; Proteins; Proteomics; Rattus; Role; Sclerosis; Serum; Smooth Muscle Myocytes; Solid; Testing; Tissues; Transplant Recipients; Transplantation; Vascular Diseases; Viral; Viral Genes; Virus; Wound Healing; angiogenesis; cytokine; heart allograft; in vitro Assay; macrophage; migration; monocyte; mutant; paracrine; restenosis; tool; virus genetics

DESCRIPTION (provided by applicant): 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). HCMV is associated with TVS and CR in solid organ transplant patients and we have observed that RCMV significantly accelerates the development of TVS and CR in heart allografts in the rat transplant model. Although we can detect the presence of virus in the transplanted heart throughout the disease course, the number of infected cells in the allograft does not account for the global effects of RCMV infection of the organ in the development of TVS. This lack of correlation suggests that infected cells are influencing the microenvironment via paracrine mechanisms. Analysis of allografts with RCMV-accelerated TVS for host cell gene expression by microarray analysis revealed a significant number of genes involved in tissue remodeling (WH) and angiogenesis (AG) that were highly up-regulated in the organ transplant. In addition our preliminary results indicate that virus-free and serum-free cellular supernatants obtained from HCMV- but not mock- infected cells induce both WH & AG in in vitro assays. Mass Spec analysis of the HCMV secretome identified multiple angiogenic and WH agonists and antagonists that most likely contribute to the tissue remodeling processes. We hypothesize that the spectrum of cytokines and growth factors secreted by CMV-infected cells significantly contribute to the acceleration of TVS and CR in heart allografts through the stimulation of WH and AG. Therefore, in this project we propose the use of viral genetics to unravel the WH and AG mechanisms involved in CMV acceleration of TVS and CR. In the first specific aim of this project we will use HCMV WT and mutant viruses as tools in combination with gel-free LC Mass Spec proteomics and in vitro WH and AG assays to identify specific viral genes and cellular proteins as well as the mechanisms through which viral genes induce tissue-remodeling events. In the second aim of this project we will knock-out the HCMV correlate gene(s) in RCMV and test this virus(es) in WH and AG assays, as well as analyze the viral secretome(s) by Mass Spec analysis. Lastly, we will examine the effect of these mutations on the ability of the virus to accelerate TVS in the RCMV rat heart transplant model. The anticipation is that multiple HCMV genes are responsible for the induction of WH and AG and that we will be able to separate some of these processes to examine the effect of mutation of correlate RCMV genes in the acceleration of TVS.

描述（由申请人提供）：该项目的长期目标是确定人类巨细胞病毒（HCMV）在加速血管疾病（例如动脉粥样硬化，再狭窄和移植血管硬化症（TVS））中的作用。 HCMV与固体器官移植患者的TVS和CR相关，我们观察到RCMV在大鼠移植模型中显着加速了TVS和CR的发育。尽管我们可以在整个疾病过程中检测到移植心脏中病毒的存在，但同种异体移植中感染细胞的数量并不能解释器官RCMV感染在电视开发中的全球影响。缺乏相关性表明，受感染的细胞正在通过旁分泌机制影响微环境。通过微阵列分析对使用RCMV加速电视进行宿主细胞基因表达的同种异体移植物的分析表明，在器官移植中高度上调了组织重塑（WH）和血管生成（WH）和血管生成（AG）的大量基因。此外，我们的初步结果表明，从HCMV-但未模拟感染的细胞获得的无病毒和无血清细胞上清液会在体外测定中诱导WH＆AG。 HCMV分泌组的质量分析确定了多种血管生成和WH激动剂以及拮抗剂，这些血管生成很可能有助于组织重塑过程。我们假设CMV感染细胞分泌的细胞因子和生长因子的光谱通过刺激WH和AG显着促进了心脏同种异体移植物中TV和CR的加速。因此，在这个项目中，我们建议使用病毒遗传学来揭示与电视和CR的CMV加速有关的WH和Ag机制。在该项目的第一个具体目的中，我们将使用HCMV WT和突变病毒作为工具与无凝胶LC质量质量蛋白质组学以及体外WH和AG分析相结合，以鉴定特定的病毒基因和细胞蛋白，以及病毒基因诱导组织造成组织造成的机制。在该项目的第二个目标中，我们将在RCMV中淘汰HCMV相关基因，并在WH和AG分析中测试该病毒（ES），并通过质量规格分析分析病毒秘密组。最后，我们将研究这些突变对病毒在RCMV大鼠心脏移植模型中加速电视的能力的影响。人们期望的是，多个HCMV基因是WH和AG诱导的原因，并且我们将能够分离其中的某些过程，以检查RCMV基因突变在TVS加速中的突变。

项目成果

HCMV pUS28 initiates pro-migratory signaling via activation of Pyk2 kinase.

• DOI: 10.1186/2042-4280-1-2
• 发表时间: 2010-12-07
• 期刊: Herpesviridae
• 作者: Vomaske J;Varnum S;Melnychuk R;Smith P;Pasa-Tolic L;Shutthanandan JI;Streblow DN
• 通讯作者: Streblow DN