Effects of human cytomegalovirus on monocyte survival and differentiation

人巨细胞病毒对单核细胞存活和分化的影响

基本信息

批准号：
8894196
负责人：
Gary Ching Tao Chan
金额：
$ 40.25万
依托单位：
UPSTATE MEDICAL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-08-01 至 2017-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8894196
关键词：
Acquired Immunodeficiency Syndrome Acute Antiviral Agents Apoptosis Apoptotic Atherosclerosis BCL2 gene Back Binding Blood Circulation Bypass Cell Death Cell Differentiation process Cells Chronic Co-Immunoprecipitations Cytomegalovirus Cytomegalovirus Infections Development Differentiation Inducer Disease Epidermal Growth Factor Receptor Etiology Family Family member Genes Genetic Transcription Glycoproteins HSPB1 gene Health Heat Shock Protein 27 Hematogenous Hour Immunocompetent Immunocompromised Host Individual Infection Infiltration Inflammation Inflammatory Inflammatory Bowel Diseases Integrins Lead Life Ligands Mass Spectrum Analysis Mediating MicroRNAs Modeling Molecular Molecular Mechanisms of Action Morbidity - disease rate Myelogenous Myeloid Cells Nuclear Translocation Organ Organ failure Output Pathogenesis Patients Peripheral Pharmacologic Substance Phosphatidylinositols Phosphotransferases Play Process Protein Family Proteins Regulation Role Signal Pathway Signal Transduction Site Staging Staining method Stains Testing Therapeutic Time Tissues Transplant Recipients Use of New Techniques Viral Viral Physiology Viral Proteins Virus Virus Replication annexin A5 caspase-3 cell growth regulation cellular targeting design immunosuppressed inhibitor/antagonist inositol-1,4,5-trisphosphate 5-phosphatase leukemia macrophage meetings monocyte mortality mutant neonate neutralizing antibody novel protein expression protein protein interaction receptor research study small molecule

项目摘要

DESCRIPTION (provided by applicant): Human cytomegalovirus (HCMV) infection is generally asymptomatic in immunocompetent individuals, although HCMV is a primary viral candidate in the etiology of several chronic inflammatory diseases including atherosclerosis and inflammatory bowel disease. In immunocompromised individuals, such as neonates, AIDS patients, and transplant recipients, HCMV infection can lead to acute multi-organ inflammation resulting in significant morbidity and mortality. Inflammatory organ diseases associated with a HCMV infection is a direct consequence of the systemic viral spread to and infection of multiple organ sites that occur during either asymptomatic or symptomatic infections. Monocytes are responsible for delivering the virus into tissues and play a central role in the inflammatory state of infected organs. However, because anti-apoptotic viral proteins are not expressed during the early stages of infection, it remains unclear how HCMV simultaneously promotes the survival and pro-inflammatory differentiation of these short-lived cells. We hypothesize that HCMV stimulates a unique cellular anti-apoptotic reprogramming specifically designed to meet the viability needs of differentiating infected monocytes, which will be tested in 3 separate aims. Aim 1 will delineate which viral glycoprotein and cellular receptor interactions occurring during viral entry are responsible for the rapid block of monocyte apoptosis. Neutralizing antibodies and pharmaceutical inhibitors will be used individually or in different combinations to dissect the rol of each viral ligand and cellular receptor interaction in promoting the survival of HCMV-infected monocytes. Aim 2 will determine how HCMV concurrently drives the long-term survival and the pro-inflammatory differentiation of infected monocytes. Mutant viruses and anti-miRs [bind and inhibit microRNAs (miRNAs)] will be used to neutralize the activity of viral and cellular miRNAs during infection of monocytes. Monocyte survival and differentiation will then be examined by Annexin V staining and Affymetrix microarray gene analysis, respectively. Aim 3 will identify the molecular mechanism of action and therapeutic potential of targeting cellular anti-apoptotic Bcl-2 family proteins induced during HCMV infection of monocytes. To define the mode of action of virally induced cellular anti-apoptotic proteins, co-immunoprecipitation experiments will be done along with mass spectrometry to globally identify new and unique protein-protein interactions specific to HCMV-infected monocytes. BH3 profiling, a new technique used for predicting the sensitivity of cells to small-molecule inhibitors against anti-apoptotic Bcl-2 family members, will be used to determine the antiviral potential of targeting cellular Bcl-2 family survival proteins i order to direct infected monocytes towards cell death. These studies will increase our understanding on the pathogenesis and dissemination of HCMV, evaluate new antiviral strategies that target infected cells rather than the virus directly, and provide a proof-of-princile for the use of BH3 profiling to predict the efficacy of Bcl- 2 family small-molecule antagonists at eliminating specific virus and cell infection combinations.

描述（由申请人提供）：尽管HCMV是多种慢性炎症性疾病的病因，包括动脉粥样硬化和炎症性肠病，但在免疫能力的个体中，人类巨细胞病毒（HCMV）感染通常是无症状的。在免疫功能低下的个体（例如新生儿，AIDS患者和移植受者）中，HCMV感染会导致急性多器官炎症，从而导致明显的发病率和死亡率。与HCMV感染相关的炎症器官疾病是全身病毒传播到无症状或有症状感染过程中多个器官部位的全身病毒传播和感染的直接结果。单核细胞负责将病毒输送到组织中并在炎症状态下起核心作用感染器官。但是，由于在感染的早期阶段未表达抗凋亡病毒蛋白，因此尚不清楚HCMV如何同时促进这些短寿命细胞的生存和促炎分化。我们假设HCMV刺激专门设计的独特的细胞抗凋亡重编程，以满足区分受感染的单核细胞的可行性需求，这将在3个单独的目标中进行测试。目的 1将描述在病毒期间发生哪种病毒糖蛋白和细胞受体相互作用进入是单核细胞凋亡的快速块。中和抗体和药物抑制剂将单独或以不同的组合使用，以剖析每种病毒配体的ROL和细胞受体相互作用，以促进HCMV感染的单核细胞的存活。 AIM 2将确定HCMV如何同时驱动感染单核细胞的长期生存和促炎性分化。突变病毒和抗MIRS [结合和抑制microRNA（miRNA）]将用于中和单核细胞感染期间病毒和细胞miRNA的活性。然后，将分别通过膜联蛋白V染色和Affymetrix微阵列基因分析来检查单核细胞的存活和分化。 AIM 3将确定靶向单核细胞HCMV感染期间诱导的细胞抗凋亡Bcl-2家族蛋白的作用和治疗潜力的分子机制。为了定义病毒诱导的细胞抗凋亡蛋白的作用方式，将与质谱法一起进行共免疫沉淀实验，以识别全球范围内针对HCMV感染的单核细胞的新的和独特的蛋白质 - 蛋白质相互作用。 BH3分析是一种用于预测细胞对小分子抑制剂对抗凋亡Bcl-2家族成员的敏感性的新技术，将用于确定靶向细胞Bcl-2家族存活蛋白I下令将感染单核细胞转向细胞死亡的抗病毒潜力。这些研究将提高我们对HCMV的发病机理和传播的理解，评估靶向感染细胞而不是直接靶向感染细胞的新抗病毒药策略，并为使用BH3分析提供了预测BCL-2家族小分子拮抗剂在使用BH3分析的有效性的原则证明。消除特定的病毒和细胞感染组合。