Molecular Mechanisms and In Vivo Impact of Tethered Mucin 1-Influenza Virus Interactions

系留粘蛋白 1-流感病毒相互作用的分子机制和体内影响

• 批准号: 10543836
• 负责人: Margaret Adele Scull
• 金额: $ 56.83万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-15 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10543836
• 关键词: Address; Affect; Age; Amino Acids; Apical; Atlases; Bacterial Infections; Binding; Biological Assay; Biological Models; Biology; Bone Marrow; Cell Compartmentation; Cell Culture Techniques; Cell surface; Cells; Cellular Structures; Clinical; Consensus; Cytoplasmic Tail; Data; Disease; Epidemic; Epithelial Cells; Flow Cytometry; Gel; Glycoproteins; Goals; Hematopoietic; Human; Immune response; Immunohistochemistry; In Vitro; Individual; Infection; Inflammation; Influenza A virus; Interferons; Investigation; Kinetics; Knockout Mice; Laboratories; Lung; Macrophage; Malignant Neoplasms; Mass Spectrum Analysis; Mediation; Modeling; Molecular; Morbidity - disease rate; Mucin 1 protein; Mucins; Mucous body substance; Mus; Nature; Outcome; Pathogenesis; Pathogenicity; Penetration; Persons; Phosphorylation; Physiological; Population; Proteins; Public Health; Reporting; Research; Respiratory System; Role; Sialic Acids; Signal Transduction; Signaling Molecule; Stable Isotope Labeling; Surface; System; Technology; Testing; Time; Tracheal Epithelium; Tumor Promotion; Up-Regulation; Viral; Viral Pathogenesis; Virus Diseases; Virus Replication; acute infection; airway epithelium; cell type; defined contribution; glycosylation; in vivo; in vivo Model; influenzavirus; innovation; insight; mortality; mouse model; novel; pandemic disease; pathogen; pathogenic virus; posttranscriptional; preference; respiratory pathogen; therapeutic target; tumor progression; uptake

ABSTRACT To infect airway epithelial cells, influenza A virus (IAV) must penetrate the secreted mucus and underlying periciliary layer barriers – both comprised, in large part, of mucin glycoproteins that can be secreted or tethered to the apical surface of cells. The importance of tethered mucins in various disease states and acute infection is increasingly being realized. Specifically, tethered mucin 1 (MUC1) has been implicated in various aspects of both bacterial and viral infections. However, these reports do not yield consensus on MUC1 function during pathogenic insult and while it seems logical to assume that MUC1 expression would benefit the host, our preliminary data indicate that MUC1 facilitates IAV infection. As MUC1 is expressed by multiple cell types that influence IAV pathogenesis, including airway epithelial cells and macrophages, and has been proposed to act as both a structural barrier and signaling molecule, we hypothesize that MUC1 facilitates IAV infection through multiple mechanisms, involving both physical and functional attributes of MUC1, and multiple cell types. We will test this hypothesis by pursing two specific aims: 1) Define the contribution of MUC1 expression in specific cell compartments to IAV pathogenesis and 2) Define MUC1 molecular interactions and intracellular dynamics in human airway epithelium during IAV infection. Aim 1 will generate a MUC1-expression atlas in the lung during infection, determine the impact of MUC1 expression on IAV infection of both human and mouse primary airway epithelial cells, and define the contribution of MUC1 expression in specific cell compartments to the outcome of infection in vivo. Aim 2 will interrogate the physical interaction between IAV and MUC1 in physiologically relevant models of airway epithelium and define the impact of IAV and interferon on MUC1 expression, activation, and interacting partners through mechanistic studies and comprehensive mass spectrometry-based technology. This research is innovative in its use of novel in vitro and in vivo models with altered MUC1 expression to dissect the interplay between IAV and MUC1 at both the molecular and organismal level. Our results will be significant as defining MUC1 function in the context of IAV infection will advance our understanding of tethered mucin biology and reveal mechanisms of how IAV negotiates specific mucin molecules to efficiently infect the respiratory tract.

抽象的 为了感染气道上皮细胞，甲型流感病毒 (IAV) 必须穿透分泌的粘液和底层 纤毛周围层屏障——两者大部分都由可以分泌或束缚的粘蛋白糖蛋白组成 至细胞顶面。束缚粘蛋白在各种疾病状态和急性感染中的重要性 正在被越来越多地认识到。具体而言，束缚粘蛋白 1 (MUC1) 与 细菌和病毒感染。然而，这些报告并未就 MUC1 的功能达成共识。 致病性损伤，虽然假设 MUC1 表达有利于宿主似乎是合乎逻辑的，但我们的 初步数据表明MUC1促进IAV感染。由于 MUC1 由多种细胞类型表达， 影响 IAV 发病机制，包括气道上皮细胞和巨噬细胞，并已被提议发挥作用 作为结构屏障和信号分子，我们假设 MUC1 通过以下方式促进 IAV 感染： 多种机制，涉及 MUC1 的物理和功能属性以及多种细胞类型。我们 将通过追求两个具体目标来检验这一假设：1) 定义 MUC1 表达在特定情况下的贡献 细胞区室对 IAV 发病机制的影响以及 2) 定义 MUC1 分子相互作用和细胞内动力学 IAV 感染期间人类气道上皮细胞中。目标 1 将在肺部生成 MUC1 表达图谱 在感染过程中，确定 MUC1 表达对人和小鼠原代 IAV 感染的影响 气道上皮细胞，并定义特定细胞区室中 MUC1 表达对气道上皮细胞的贡献 体内感染的结果。目标 2 将询问 IAV 和 MUC1 之间的物理相互作用 气道上皮的生理相关模型并定义 IAV 和干扰素对 MUC1 的影响 通过机制研究和综合质量来表达、激活和相互作用伙伴 基于光谱测量的技术。这项研究的创新之处在于使用了新颖的体外和体内模型 改变 MUC1 表达以剖析 IAV 和 MUC1 之间的分子和分子相互作用 有机体水平。我们的结果将具有重要意义，因为在 IAV 感染的背景下定义 MUC1 功能将 增进我们对系留粘蛋白生物学的理解，并揭示 IAV 如何协商特定的机制 粘蛋白分子有效感染呼吸道。