HCMV receptors, signaling and entry

HCMV 受体、信号传导和进入

• 批准号: 10669505
• 负责人: TIMOTHY F KOWALIK
• 金额: $ 53.17万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-06 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10669505
• 关键词: Address; Adolescent; Amino Acids; Binding; CD46 Antigen; CRISPR screen; CRISPR/Cas technology; Cell Nucleus; Cell physiology; Cells; Clinical; Clinical Pathology; Complex; Congenital Abnormality; Cryoelectron Microscopy; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Cytomegalovirus; Data; Disease; Endothelial Cells; Epidermal Growth Factor Receptor; Epithelial Cells; Family; G-Protein-Coupled Receptors; GTP-Binding Protein alpha Subunits, Gs; GTP-Binding Proteins; Genes; Glycoproteins; Goals; Hematopoietic; Herpesviridae; Human; Immune system; Immunocompromised Host; Individual; Infant; Infection; Integration Host Factors; Integrins; Knowledge; Life Cycle Stages; Link; Longitudinal Studies; Malignant Glioma; Malignant Neoplasms; Mediating; Membrane Proteins; Microscopy; Molecular; Molecular Genetics; Molecular Target; Morbidity - disease rate; N-terminal; Nature; Neuropilin-2; Orphan; Pathogenesis; Pathway interactions; Pattern; Peptides; Persons; Play; Population; Pregnancy Complications; Process; Protein Tyrosine Kinase; Proteins; Receptor Cell; Receptor Signaling; Reporting; Research; Role; Sensorineural Hearing Loss; Signal Pathway; Signal Transduction; Signaling Molecule; Source; Stream; Structure; Testing; Tropism; Viral; Virus; Virus Diseases; Virus Replication; Work; base; base editing; cell type; clinically relevant; cofactor; cytomegalovirus receptor; druggable target; extracellular; immunosuppressed; member; monocyte; mortality; new therapeutic target; novel; olfactory receptor; peptidomimetics; receptor; screening; trafficking; young adult

PROJECT SUMMARY/ABSTRACT Human cytomegalovirus (HCMV) is the leading infectious cause of birth defects in infants. HCMV can also cause a variety of severe diseases in immunocompromised individuals. Mounting evidence have linked HCMV infections to some cancers, most notably malignant glioma. Major progress has been made during the last decade in understanding the molecular mechanism of viral entry and replication. A variety of host receptors and signaling molecules have been shown to regulate HCMV infection. Recently, we used a CRISPR/Cas9 gene editing based screening strategy to search for cellular proteins that are required for HCMV infection of epithelial cells. We found that silencing the expression of an orphan olfactory receptor, OR14I1, drastically reduced HCMV infection in these cells. Further work showed that OR14I1 interacts with the HCMV Pentamer complex (PC). Together with additional evidence, we confirmed that OR14I1 is an HCMV receptor and may play critical role in defining PC-dependent tropism. Despite the progress of identification of several host receptors for HCMV, there are several critical gaps in our current knowledge of the molecular mechanism of viral entry and subsequent viral life cycle. Specifically, it is unclear how HCMV uses multiple host receptors for entry and how virus subvert host cellular signaling pathways facilitate viral replication and spreading. Our long-term goals are to identify how HCMV uses normal host cellular processes to facilitate viral infection, to elucidate the entry mechanism related to viral tropism, define the entry signaling pathways, and trafficking of HCMV to the nucleus by clarifying the roles of OR14I1 and other receptors associated with its broad tropism and pathogenesis. Building on the past work of our group and others, we propose to conduct research on three Specific Aims to address these goal: (1) To elucidate the signaling pathways activated by HCMV-OR14I1 binding and their roles in virus infection and uncover the role OR14I1 plays in cell-cell spread and trafficking; (2) To determine the contribution of different HCMV PC receptors to infection and tropism; and (3) determine the structure of HCMV PC-OR14I1 and determine relevant molecular interactions. Collectively, our proposed research will broadly impact the field by characterizing the essential roles that host receptors, in particular, OR14I1, play in promoting viral entry, replication, and spreading. These studies will have the potential to uncover novel molecular mechanisms underlying HCMV infection, and molecular targets for HCMV therapy.

项目摘要/摘要 人类巨细胞病毒（HCMV）是婴儿出生缺陷的主要感染原因。 HCMV也可能导致 免疫功能低下的个体中有多种严重疾病。越来越多的证据与HCMV联系在一起 对某些癌症的感染，最著名的是恶性神经胶质瘤。最后的取得了重大进展 了解病毒进入和复制的分子机制的十年。各种宿主受体和 信号分子已显示出调节HCMV感染。最近，我们使用了CRISPR/CAS9基因 基于编辑的筛选策略以搜索上皮感染所需的细胞蛋白 细胞。我们发现沉默的孤儿嗅觉受体OR14I1的表达大大降低了HCMV 这些细胞中的感染。进一步的工作表明，OR14I1与HCMV Pentamer复合物（PC）相互作用。 与其他证据一起，我们确认OR14I1是HCMV受体，可能在 定义依赖于PC的偏向主义。尽管鉴定了几种HCMV宿主受体的鉴定，但 在我们目前对病毒进入分子机制和随后的病毒的了解中，这是几个关键的差距 生命周期。具体而言，目前尚不清楚HCMV如何使用多个宿主受体入口以及病毒颠覆宿主 细胞信号通路有助于病毒复制和扩散。我们的长期目标是确定如何 HCMV使用正常的宿主细胞过程来促进病毒感染，以阐明与入口机制相关 为了病毒朝向主义，通过澄清hCMV来定义进入信号通路以及将HCMV运输到细胞核上 OR14I1的作用以及其他与其广泛的巨型和发病机理相关的受体。过去建立 我们小组和其他人的工作，我们建议对三个特定目的进行研究以解决这些目标：（1） 阐明由HCMV-OR14I1结合激活的信号通路及其在病毒感染中的作用 揭示OR14I1在细胞传播和贩运中的作用； （2）确定不同的贡献 HCMV PC受体受感染和巨型主义的受体； （3）确定HCMV PC-OR14I1和 确定相关的分子相互作用。总的来说，我们拟议的研究将通过 表征宿主受体特别是OR14I1在促进病毒进入中发挥的基本作用， 复制和传播。这些研究将有可能发现新颖的分子机制 基础HCMV感染和HCMV治疗的分子靶标。