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

• 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
• 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

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（包括动脉粥样硬化和炎症性肠病）病因学中的主要候选病毒，但在免疫功能正常的个体中，人巨细胞病毒 (HCMV) 感染通常是无症状的。在免疫功能低下的个体中，例如新生儿、艾滋病患者和移植受者，HCMV 感染可导致急性多器官炎症，导致显着的发病率和死亡率。与 HCMV 感染相关的炎症器官疾病是无症状或有症状感染期间发生的全身性病毒扩散到多个器官部位并感染的直接后果。单核细胞负责将病毒输送到组织中，并在炎症状态中发挥核心作用 受感染的器官。然而，由于抗凋亡病毒蛋白在感染早期并不表达，因此目前尚不清楚 HCMV 如何同时促进这些短命细胞的存活和促炎分化。我们假设 HCMV 会刺激一种独特的细胞抗凋亡重编程，这种重编程是专为满足区分受感染单核细胞的活力需求而设计的，这将在 3 个不同的目标中进行测试。目的 图1将描述病毒感染过程中发生的病毒糖蛋白和细胞受体相互作用 进入负责快速阻止单核细胞凋亡。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和细胞受体相互作用在促进 HCMV 感染的单核细胞存活中的作用。目标 2 将确定 HCMV 如何同时驱动受感染单核细胞的长期存活和促炎分化。突变病毒和抗 miR [结合并抑制 microRNA (miRNA)] 将用于中和单核细胞感染期间病毒和细胞 miRNA 的活性。然后分别通过膜联蛋白 V 染色和 Affymetrix 微阵列基因分析检查单核细胞存活和分化。目标 3 将确定单核细胞 HCMV 感染期间诱导的靶向细胞抗凋亡 Bcl-2 家族蛋白的分子作用机制和治疗潜力。为了确定病毒诱导的细胞抗凋亡蛋白的作用模式，将与质谱一起进行免疫共沉淀实验，以全面鉴定针对 HCMV 感染的单核细胞的新的、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抗凋亡 Bcl-2 家族成员的小分子抑制剂敏感性的新技术， 可用于确定靶向细胞 Bcl-2 家族存活蛋白的抗病毒潜力，以引导受感染的单核细胞死亡。这些研究将增进我们对 HCMV 发病机制和传播的理解，评估针对受感染细胞而不是直接针对病毒的新抗病毒策略，并为使用 BH3 分析预测 Bcl-2 家族小分子拮抗剂的功效提供原理证明。 消除特定的病毒和细胞感染组合。