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

摘要 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下面的上皮细胞。 纤毛周层屏障-两者都在很大程度上由粘蛋白糖蛋白组成， 到细胞的顶端表面。栓系粘蛋白在各种疾病状态和急性感染中的重要性 越来越多地被认识到。具体地，栓系粘蛋白1（MUC 1）已经涉及免疫调节的各个方面。 包括细菌和病毒感染。然而，这些报道并没有产生关于MUC 1功能的共识， 虽然假设MUC 1表达对宿主有益似乎是合乎逻辑的，但我们的研究表明， 初步数据表明MUC 1促进IAV感染。由于MUC 1由多种细胞类型表达， 影响IAV的发病机制，包括气道上皮细胞和巨噬细胞，并已被提出发挥作用 作为一种结构屏障和信号分子，我们假设MUC 1通过以下途径促进IAV感染： 多种机制，涉及MUC 1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）确定MUC 1表达在特定细胞中的作用， 2）定义MUC 1分子相互作用和细胞内动力学 在人呼吸道上皮中的作用。Aim 1将在肺中生成MUC 1表达图谱 在感染过程中，确定MUC 1表达对IAV感染人和小鼠原发性 气道上皮细胞，并确定MUC 1表达的贡献，在特定的细胞区室， 体内感染的结果。目的2将询问IAV和MUC 1之间的物理相互作用， 呼吸道上皮的生理相关模型，并确定IAV和干扰素对MUC 1的影响 表达，激活和相互作用的合作伙伴，通过机制的研究和全面的质量 基于光谱的技术。这项研究在使用新型体外和体内模型方面具有创新性， 改变MUC 1的表达，以在分子水平和细胞水平上分析IAV和MUC 1之间的相互作用。 有机体水平。我们的结果将是重要的，因为在IAV感染的背景下定义MUC 1功能将 推进我们对系留粘蛋白生物学的理解，并揭示IAV如何谈判特异性 粘蛋白分子有效地感染呼吸道。

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.