Human cytomegalovirus-induced alterations to cell-surface adhesion proteins

人巨细胞病毒诱导的细胞表面粘附蛋白改变

• 批准号: 8885082
• 负责人: THOMAS E SHENK
• 金额: $ 16.26万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8885082
• 关键词: Address; Adhesions; Adhesives; Adult; Area; Autocrine Communication; Biology; Blindness; Breast; C-terminal; Cell Adhesion; Cell Adhesion Molecules; Cell Communication; Cell Nucleus; Cell Surface Proteins; Cell surface; Cells; Clinical; Congenital Abnormality; Cytomegalovirus; Cytomegalovirus Infections; Cytomegalovirus Retinitis; DNA Damage; Disease; Environment; Epithelial Cells; Extracellular Domain; Family; Family member; Fibroblasts; Foundations; G-Protein-Coupled Receptors; Gene Expression; Human; Immunosuppression; Individual; Infection; Kidney; Laboratories; Lead; Life; Mass Spectrum Analysis; Mediating; Mental Retardation; N-terminal; Neoplasm Metastasis; Nervous system structure; Neural Cell Adhesion Molecule L1; Neurites; Nuclear; Outcome; Paracrine Communication; Pathogenesis; Pharmaceutical Preparations; Play; Population; Prevalence; Production; Proteins; Proteome; Proteomics; Radiation Tolerance; Radioresistance; Role; SHFM1 gene; Salivary Glands; Signal Transduction; Signaling Molecule; Site; Testing; Viral; Viral Pathogenesis; base; cell behavior; cell motility; cell type; congenital infection; design; disability; extracellular; hearing impairment; immunosuppressed; insight; member; public health relevance; response; tumor; tumor progression

 DESCRIPTION (provided by applicant): Human cytomegalovirus (HCMV) infects over 60% of the adult population. It is a major cause of birth defects, and a life-threatening opportunistic agent in immunosuppressed people. Congenital infection has a significant prevalence and can cause permanent disabilities. Proteomic analysis in our lab has recently revealed that numerous cell-surface proteins are dynamically modulated following HCMV infection of fibroblasts, including 27 cell-surface adhesion proteins. Two upregulated adhesion proteins, L1CAM and GPR56, have been tested further and shown to significantly impact the yield of HCMV progeny. These observations form the foundation for the central hypothesis underlying this application: multiple cellular adhesion proteins cooperate to modulate the outcome of HCMV infection. To address this hypothesis, we will first study the IgCAM family member, L1CAM, which is elevated in a wide range of human tumors, inducing radioresistance and often enhancing tumor progression or metastasis. Our preliminary studies show that cell-surface L1CAM is elevated by infection and supports the production of HCMV progeny in fibroblasts. Secreted L1CAM extracellular domain (L1CAM-ECD, sponsors autocrine and paracrine signal transduction) and nuclear L1CAM intracellular domain (L1CAM-ICD modulates cellular gene expression) also are dramatically increased following infection. Further, knockdown of L1CAM sensitized infected cells to radiomimetic drugs. The remarkable range of activities documented for L1CAM provide a compelling rationale for detailed study of its role in HCMV replication and spread, i.e., L1CAM is likely to impact HCMV replication and spread at multiple levels: a direct effect on the production of HCMV progeny; and possible effects on viral spread via infected-cell migration, the infected-cell microenvironment via L1CAM-ECD activity, and the radiosensitivity/DNA-damage response of infected cells. We also will investigate GPR56, a member of the adhesion family of G protein-coupled receptors. Our initial studies of the HCMV-infected cell proteome used fibroblasts, which are infected at numerous sites within the body and serve as a reference, since they are the common laboratory host cell for HCMV studies. We will next characterize epithelial cells to expand our understanding of the infected cell-surface proteome and identify further candidates for detailed studies such as those proposed above for L1CAM. Epithelial cells are central to HCMV pathogenesis, because they are infected at the site of entry, they host long-term low-level replication in the salivary gland, breast and kidney to mediate HCMV spread to new hosts, and they are the target cell in CMV retinitis. Collectively, this study will provide he first broad-based, mechanistic characterization of adhesion proteins during HCMV infection. Successful completion of these studies should contribute importantly to the understanding of a relatively understudied area of HCMV biology, the role of cellular adhesion proteins in viral replication and dissemination.

 描述（由适用提供）：超过60％的成人人群的人类巨细胞病毒（HCMV）感染。这是造成先天缺陷的主要原因，也是免疫抑制人士中危及生命的无人选择的代理。先天性感染的患病率很高，可能导致永久性残疾。我们实验室中的蛋白质组学分析最近显示，在HCMV感染成纤维细胞（包括27个细胞表面粘附蛋白）之后，许多细胞表面蛋白是动态调节的。已经对两种上调的粘合蛋白L1CAM和GPR56进行了进一步测试，并显示出显着影响HCMV后代的产量。这些观察结果构成了此应用的基础：多个细胞粘合蛋白配合以调节HCMV感染的结果。为了解决这一假设，我们将首先研究IGCAM家族成员L1CAM，该成员在广泛的人类肿瘤中升高，诱导辐射耐药性并经常增强肿瘤进展或转移。我们的初步研究表明，细胞表面L1CAM通过感染升高，并支持成纤维细胞中HCMV进展的产生。在感染后，分泌的L1CAM细胞外结构域（L1CAM-ECD，赞助商自分泌和旁分泌信号转导）和核L1CAM细胞内结构域（L1CAM-ICD调节细胞基因表达）也会大大增加。此外，敲低L1CAM敏感的感染细胞对放射性药物。记录在L1CAM的活动范围的范围为详细研究其在HCMV复制和传播中的作用提供了令人信服的理由，即L1CAM可能会影响HCMV复制并以多个层面的范围影响：直接影响HCMV后代的产生；以及通过感染细胞迁移，通过L1CAM-ECD活性感染细胞微环境以及感染细胞的放射敏感/DNA破坏反应的可能影响。我们还将研究GPR56，这是G蛋白偶联受体的粘附家族的成员。我们对HCMV感染的细胞蛋白质组的初步研究使用了成纤维细胞，这些成纤维细胞在体内的许多部位被感染并作为参考，因为它们是HCMV研究的常见实验室宿主细胞。接下来，我们将表征上皮细胞，以扩展我们对感染的细胞表面蛋白质组的理解，并确定更多详细研究的候选者，例如上述L1CAM。上皮细胞是HCMV发病机理的核心，因为它们在入口部位被感染，因此它们在唾液腺，乳腺癌和肾脏中占据长期的低水平复制，以介导HCMV扩散到新宿主，并且它们是CMV视网膜炎的靶细胞。总的来说，这项研究将提供HCMV感染期间粘合剂蛋白的首个基于广泛的机械表征。这些研究的成功完成应重要地有助于理解相对了解的HCMV生物学领域，即细胞粘合剂蛋白在病毒复制和传播中的作用。