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.

项目概要/摘要 人类巨细胞病毒（HCMV）是导致婴儿出生缺陷的主要原因。 HCMV还可引起 免疫功能低下个体的各种严重疾病。越来越多的证据表明 HCMV 与 感染某些癌症，尤其是恶性神经胶质瘤。过去一年取得了重大进展 十年来了解病毒进入和复制的分子机制。多种宿主受体和 信号分子已被证明可以调节 HCMV 感染。最近，我们使用了 CRISPR/Cas9 基因 基于编辑的筛选策略，寻找 HCMV 感染上皮细胞所需的细胞蛋白 细胞。我们发现沉默孤儿嗅觉受体 OR14I1 的表达可显着降低 HCMV 这些细胞的感染。进一步的研究表明 OR14I1 与 HCMV 五聚体复合物 (PC) 相互作用。 结合其他证据，我们证实 OR14I1 是 HCMV 受体，可能在 定义PC依赖性向性。尽管 HCMV 的几种宿主受体的鉴定取得了进展，但 我们目前对病毒进入和随后病毒传播的分子机制的了解存在几个关键差距 生命周期。具体来说，尚不清楚HCMV如何利用多个宿主受体进入以及病毒如何颠覆宿主 细胞信号通路促进病毒复制和传播。我们的长期目标是确定如何 HCMV利用正常的宿主细胞过程来促进病毒感染，阐明相关的进入机制 通过阐明病毒趋向性、定义进入信号通路以及 HCMV 向细胞核的运输 OR14I1 和其他受体的作用与其广泛的趋向性和发病机制相关。立足于过去 根据我们小组和其他人的工作，我们建议对三个具体目标进行研究以实现这些目标：（1） 阐明 HCMV-OR14I1 结合激活的信号通路及其在病毒感染和感染中的作用 揭示 OR14I1 在细胞间传播和运输中的作用； (2) 确定不同贡献度 HCMV PC 感染和趋向性受体； (3)确定HCMV PC-OR14I1的结构和 确定相关的分子相互作用。总的来说，我们提出的研究将通过以下方式广泛影响该领域 表征宿主受体，特别是 OR14I1，在促进病毒进入中发挥的重要作用， 复制、传播。这些研究将有可能揭示新的分子机制 潜在的 HCMV 感染以及 HCMV 治疗的分子靶点。