Role of Cytomegalovirus GPCRs in Pathogenesis in Vivo

巨细胞病毒 GPCR 在体内发病机制中的作用

• 批准号: 8514752
• 负责人: WILLIAM E MILLER
• 金额: $ 30.64万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8514752
• 关键词: Acinar Cell; Acute; Adult; Affect; American; Animal Model; Animals; Antiviral Agents; Apoptosis; Area; Biological Models; Biology; Cell Survival; Cell physiology; Cells; Coupled; Cytomegalovirus; Cytomegalovirus Infections; Data; Development; Disease; Ensure; Epithelial Cells; Epithelium; Exhibits; Fetus; G-Protein Signaling Pathway; G-Protein-Coupled Receptors; Gene Expression; Genes; Genetic; Gland; Glean; Goals; Growth; Homologous Gene; Horizontal Disease Transmission; Immunocompromised Host; In Vitro; Individual; Infection; Intervention; Investigation; Knock-out; Lead; Life; Liquid substance; Liver; Molecular; Movement; Murid herpesvirus 1; Mus; Organ; Pathogenesis; Pathway interactions; Phase; Physiological; Play; Population; Property; Proteins; Public Health; Role; Saliva; Salivary; Salivary Glands; Severities; Signal Pathway; Signal Transduction; Species Specificity; Spleen; Study models; System; Testing; Tissues; Transgenic Animals; Transgenic Mice; Transgenic Organisms; United States; Viral; Viral Proteins; Virus; Virus Replication; base; cell growth; chemokine; design; in vivo; inhibitor/antagonist; innovation; insight; mouse model; mutant; novel; prevent; receptor function; research study; saliva secretion; salivary acinar cell; trafficking; transmission process

DESCRIPTION (provided by applicant): The human cytomegalovirus (HCMV) is a significant public health concern in the United States. Most important are the effects of the virus on developing fetuses and immunocompromised individuals where it causes a variety of pathological conditions ranging in severity from mild to life-threatening. Since HCMV is present in a persistent or latent form in 50-90% of the world's adult population, the identification of virl gene products that contribute to viral trafficking, persistence, and horizontal transmission is an intense and important area of investigation. Interestingly, HCMV encodes 4 genes that are homologous to cellular G- protein coupled receptors (GPCRs). The HCMV GPCRs are not essential for viral replication in vitro, however their homology to cellular GPCRs suggests that they may profoundly affect cellular physiology to ensure replication of the viruses in organs important for pathogenesis. The strict species specificity of HCMV has precluded an analysis of the function of HCMV GPCRs in vivo. However, as the biology of the related murine cytomegalovirus (MCMV) is similar to that of HCMV, the murine virus has served as a useful model for studying aspects of cytomegalovirus replication and trafficking in vivo. Interestingly, the MCMV encoded M33 GPCR is essential for replication within the salivary gland of infected mice, suggesting that M33 activity may have a direct impact on viral persistence and/or horizontal transmission of virus. Based on our preliminary data, we hypothesize that M33 signaling through the cellular Gaq/Ga11 pathway alters salivary acinar cell physiology leading to amplification and spread of the virus within the salivary gland. The proposed studies are highly significant as the salivary gland and salivary secretions play an important role in horizontal transmission of cytomegaloviruses, yet little is known about the viral properties that facilitate viral amplification within the gland and promote movement of virus into the saliva. In aim 1 we will test the hypothesis that M33 null viruses fail to amplify in the salivary gland due to defectie viral spread within the acinar epithelium. In aim 2 we will test whether M33 altered acinar cell viability or secretory activity facilitates amplification and/or spread of virus in the gland. In am 3 we will use viral and murine knockout approaches to test the hypothesis that Gaq/Ga11 is the proximal signaling pathway used by M33 for MCMV trafficking and growth in the salivary gland in vivo. We will utilize M33 knockout and wildtype viruses, knock- out and transgenic animals, and physiological analyses of salivary gland function to explore our hypothesis. The innovative in vivo experiments proposed in this application will lead to novel insights into the function of cytomegalovirus GPCRs and into mechanisms by which cytomegaloviruses persist and gain access to fluids important for horizontal transmission. Defining essential roles for cytomegalovirus GPCRs in promoting salivary gland replication and spread could ultimately lead to the development of unique antivirals designed to prevent cytomegalovirus transmission via saliva.

描述（由申请人提供）：人类巨细胞病毒（HCMV）在美国是一个重要的公共卫生问题。最重要的是该病毒对发育中的胎儿和免疫功能低下的个体的影响，在这种情况下会导致各种病理状况，从而严重程度从轻度到威胁生命。由于HCMV以持续或潜在的形式存在于世界成年人口的50-90％中，因此鉴定有助于病毒贩运，持久性和水平传播的VIRL基因产品是一个强烈而重要的调查领域。有趣的是，HCMV编码与细胞G蛋白偶联受体（GPCR）同源的4个基因。 HCMV GPCR对于体外病毒复制并不是必不可少的，但是它们与细胞GPCR的同源性表明，它们可能会深刻影响细胞生理学，以确保器官中病毒的复制对发病机理重要。 HCMV的严格物种特异性排除了对体内HCMV GPCR功能的分析。但是，由于相关鼠巨细胞病毒（MCMV）的生物学类似于HCMV，因此鼠病毒已成为研究巨细胞病毒复制和体内运输方面的有用模型。有趣的是，MCMV编码的M33 GPCR对于在感染小鼠的唾液腺内复制至关重要，这表明M33活性可能对病毒持久性和/或水平传播具有直接影响。根据我们的初步数据，我们假设通过细胞GAQ/GA11途径的M33信号传导改变了唾液腺泡细胞生理，从而导致病毒在唾液腺内的扩增和扩散。拟议的研究非常重要，因为唾液腺和唾液分泌物在巨细胞病毒的水平传播中起着重要作用，但对于促进腺体内病毒扩增并促进病毒向唾液运动的病毒特性知之甚少。在AIM 1中，我们将检验以下假设：由于腺泡上皮内缺陷病毒扩散，M33 NULL病毒无法在唾液腺中扩增。在AIM 2中，我们将测试M33是否改变了腺泡细胞活力或分泌活性是否有助于病毒在腺体中的扩增和/或传播。在AM 3中，我们将使用病毒和鼠敲除方法来检验以下假设：GAQ/GA11是M33用于MCMV运输和体内MCMV运输和生长的近端信号传导途径。我们将利用M33敲除和野生型病毒，敲除和转基因动物，以及对唾液腺功能的生理分析来探索我们的假设。本应用程序中提出的创新体内实验将导致对巨细胞病毒GPCR的功能的新见解，并使巨细胞病毒持续存在并获得对水平传播重要的液体的机制。定义巨细胞病毒GPCR在促进唾液腺复制和扩散中的基本作用最终可能导致开发旨在防止通过唾液传播的独特抗病毒药物。