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: 8990958
• 负责人: THOMAS E SHENK
• 金额: $ 39.88万
• 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/8990958
• 关键词: ADGR1 gene; 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; Health; 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; Salivary Glands; Signal Transduction; Signaling Molecule; Site; Testing; Viral; Viral Pathogenesis; Virus Replication; base; cell behavior; cell motility; cell type; congenital infection; design; disability; extracellular; hearing impairment; immunosuppressed; insight; knock-down; member; 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（由申请人提供）：人巨细胞病毒（HCMV）感染超过60%的成年人群。它是出生缺陷的主要原因，也是免疫抑制人群中危及生命的机会性因子。先天性感染发病率很高，可造成永久残疾。我们实验室的蛋白质组学分析最近揭示了许多细胞表面蛋白在HCMV感染成纤维细胞后被动态调节，包括27种细胞表面粘附蛋白。已经进一步测试了两种上调的粘附蛋白L1 CAM和GPR 56，并显示出显著影响HCMV后代的产量。这些观察结果形成了本申请的核心假设的基础：多种细胞粘附蛋白协同调节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L1 CAM，它在广泛的人类肿瘤中升高，诱导放射抗性，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L1 CAM被感染升高，并支持HCMV后代在成纤维细胞中的生产。分泌的L1 CAM胞外结构域（L1 CAM-ECD，赞助自分泌和旁分泌信号转导）和核L1 CAM胞内结构域（L1 CAM-ICD调节细胞基因表达）也在感染后显著增加。此外，L1 CAM的敲低使受感染的细胞对拟放射性药物敏感。记录的L1 CAM的显著活动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，L1 CAM可能在多个水平上影响HCMV复制和传播：对HCMV子代产生的直接影响;以及通过感染细胞迁移对病毒传播的可能影响，通过L1 CAM-ECD活性对感染细胞微环境的影响，以及感染细胞的放射敏感性/DNA损伤反应。我们还将研究GPR 56，G蛋白偶联受体的粘附家族的成员。我们对HCMV感染细胞蛋白质组的初步研究使用了成纤维细胞，其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，以扩大我们对感染细胞表面蛋白质组的理解，并确定进一步的候选人进行详细的研究，如以上提出的L1 CAM。上皮细胞是HCMV发病机制的核心，因为它们在进入位点被感染，它们在唾液腺、乳腺和肾脏中宿主长期低水平复制以介导HCMV传播到新宿主，并且它们是CMV视网膜炎的靶细胞。总的来说，这项研究将提供他的第一个广泛的，在HCMV感染的粘附蛋白的机制表征。这些研究的成功完成将有助于理解HCMV生物学的一个相对不足的领域，即细胞粘附蛋白在病毒复制和传播中的作用。