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）：人类巨细胞病毒 (HCMV) 感染超过 60% 的成年人口。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病原体。先天性感染非常普遍，可导致永久性残疾。我们实验室的蛋白质组学分析最近表明，成纤维细胞被 HCMV 感染后，许多细胞表面蛋白会发生动态调节，其中包括 27 种细胞表面粘附蛋白。两种上调的粘附蛋白 L1CAM 和 GPR56 已经过进一步测试，结果显示可显着影响 HCMV 后代的产量。这些观察结果构成了该应用的核心假设的基础：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 L1CAM，它在多种人类肿瘤中表达升高，诱导放射抗性并常常促进肿瘤进展或转移。我们的初步研究表明，细胞表面 L1CAM 因感染而升高，并支持成纤维细胞中 HCMV 后代的产生。分泌的 L1CAM 胞外结构域（L1CAM-ECD，促进自分泌和旁分泌信号转导）和核 L1CAM 胞内结构域（L1CAM-ICD 调节细胞基因表达）在感染后也会显着增加。此外，L1CAM 的敲低使受感染的细胞对放射模拟药物敏感。 L1CAM 记录的一系列显着活动为详细研究其在 HCMV 复制和传播中的作用提供了令人信服的理由，即 L1CAM 可能在多个层面上影响 HCMV 复制和传播：对 HCMV 后代产生的直接影响；以及通过受感染细胞迁移对病毒传播的可能影响，通过 L1CAM-ECD 活性对受感染细胞微环境的影响，以及受感染细胞的放射敏感性/DNA 损伤反应。我们还将研究 GPR56，它是 G 蛋白偶联受体粘附家族的成员。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，这些成纤维细胞在体内多个部位被感染，可作为参考，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，以扩大我们对受感染细胞表面蛋白质组的理解，并确定进一步的候选者进行详细研究，例如上面针对 L1CAM 提出的研究。上皮细胞是 HCMV 发病机制的核心，因为它们在进入部位被感染，它们在唾液腺、乳腺和肾脏中长期低水平复制，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，这项研究将首次提供 HCMV 感染期间粘附蛋白的广泛基础的机制表征。这些研究的成功完成应该有助于理解 HCMV 生物学中相对较少研究的领域，即细胞粘附蛋白在病毒复制和传播中的作用。