Genetic and biophysical mechanisms that control influenza virus cellular multiplicity of infection

控制流感病毒细胞感染多重性的遗传和生物物理机制

• 批准号: 10659426
• 负责人: Michael D Vahey
• 金额: $ 38.96万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-16 至 2028-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10659426
• 关键词: Address; Affect; Antiviral Response; Area; Binding Proteins; Biology; Biophysical Process; Biophysics; Cells; Cellular Tropism; Cilia; Data; Defect; Disease Progression; Disparate; Environment; Enzymes; Equilibrium; Frequencies; Genes; Genetic; Genome; Goals; Growth; Human; Image; Infection; Influenza; Integration Host Factors; Interferons; Intrinsic factor; Knowledge; Link; Mediating; Medical; Membrane Proteins; Methodology; Methods; Molecular; Movement; Mucins; Mucociliary Clearance; Mucous Membrane; Mutation; National Institute of Allergy and Infectious Disease; Neuraminidase; Outcome; Pattern; Phenotype; Play; Process; Proteins; Research; Resolution; Role; Shapes; Sialic Acids; Site; Strategic Planning; Structural Protein; Structure; Surface; Testing; Variant; Viral; Viral Load result; Viral Proteins; Virion; Virus; Virus Diseases; Virus Receptors; Virus Replication; Work; biophysical properties; co-infection; experimental study; flu transmission; high resolution imaging; improved; influenza infection; influenzavirus; insight; outcome disparities; particle; prevent; receptor; receptor binding; response; secondary infection; tool; trait; universal influenza vaccine; virus morphology

Single cells infected by influenza can produce hundreds to thousands of infectious new virions. These virions spread non-uniformly, producing wide variations in the viral load per cell that are concentrated around the initial site of infection. Differences in the amount of virus that infects a particular cell can influence whether or not that cell produces new virions of its own, or if it mounts an anti-viral response. Understanding how influenza virions spread is therefore critical to understanding how infection progresses and how the host responds. The central goal of this project is to understand how genetic and biophysical features of both virus and host contribute to the spatial structure of influenza virus cellular spread, and how differences in cellular spread shape the progression of infection and the resulting cellular responses. Our prior data demonstrate that genetic and biophysical features of influenza control the way that the virus spreads at the cellular level. These features are strongly linked to three viral proteins in particular: HA, NA, and M1. The receptor-binding protein HA mediates virus attachment to naïve cells, while the receptor-destroying protein NA facilitates virus release and dissemination. The matrix protein M1 controls the shape of the virus particle and the distribution of HA and NA on the virion surface. Collectively, these proteins control the biophysical characteristics of virus particles and shape the way that virions spread throughout the host. We hypothesize that genetic mechanisms acting through these proteins, together with host factors involved in mucociliary clearance, determine the spatial pattern of viral spread and the frequency of cellular co-infection, thereby shaping the progression of disease. We will test this hypothesis through two specific aims. In Aim 1, we will use high- resolution imaging to track the spread of virions and viral infection, and we will determine how this depends on natural variations in HA, NA, and M1. Through these experiments, we will identify how these proteins collectively influence the degree of cellular co-infection that occurs during multi-cycle virus replication. In Aim 2, we will investigate how host factors involved in mucociliary clearance contribute to cellular spread of IAV, and we will determine the collective impact of viral and host factors that alter the frequency of co-infection on key infection outcomes in differentiated human airway cells. The expected outcome of this project is an improved understanding of how influenza virus surface and structural proteins contribute to intracellular aspects of viral replication by tuning the degree of co-infection that occurs during multi-cycle growth. Insights from this work will inform basic understanding of how influenza viruses navigate the host environment and will identify host and viral factors that contribute to the disparate outcomes of infection that are sometimes observed. This proposal will also introduce new tools and methodologies for investigating the spatial organization and dynamics of influenza virus infection.

受影响力感染的单细胞可以产生数百至数千种感染的新病毒。这些病毒不均匀地扩散，在每个细胞的病毒载荷中产生广泛的变化，这些变化集中在初始感染部位周围。感染特定细胞的病毒量的差异会影响该细胞是否会产生其自身的新病毒，或者是否会安装抗病毒反应。因此，了解影响病毒传播的影响对于了解感染的进展以及宿主如何反应至关重要。该项目的核心目的是了解病毒和宿主的遗传和生物物理特征如何有助于流感病毒病毒细胞扩散的空间结构，以及细胞扩散的差异如何形成感染的进展和所得的细胞反应。我们先前的数据表明，影响力的遗传和生物物理特征控制病毒在细胞水平上传播的方式。这些特征与三种病毒蛋白特别相关：HA，NA和M1。受体结合蛋白HA介导病毒附着在幼稚的细胞上，而被受体的蛋白质NA促进了病毒释放和传播。基质蛋白M1控制病毒颗粒的形状以及HA和Na在病毒体表面上的分布。这些蛋白质共同控制病毒颗粒的生物物理特性，并塑造病毒在整个宿主中传播的方式。我们假设通过这些蛋白质作用的遗传机制，以及涉及粘膜清除率的宿主因素，确定病毒扩散的空间模式和细胞共同感染的频率，从而塑造疾病的发展。我们将通过两个具体目标检验这一假设。在AIM 1中，我们将使用高分辨率成像来跟踪病毒体和病毒感染的扩散，我们将确定这如何取决于HA，NA和M1的自然变化。通过这些实验，我们将确定这些蛋白质如何共同影响多周期病毒复制期间发生的细胞共感染程度。在AIM 2中，我们将研究涉及粘膜缩减清除率的宿主因素如何有助于IAV的细胞扩散，我们将确定病毒和宿主因子的集体影响，这些因素会改变共同感染对分化人类气道细胞中关键感染结果的频率。该项目的预期结果是对影响纳的影响力表面和结构蛋白如何通过调整多周期生长过程中发生的共感染程度来促进病毒复制的细胞内方面的预期了解。这项工作的见解将为您如何导航宿主环境导航，并确定宿主和病毒因素有时​​有时会被观察到的宿主和病毒因素。该建议还将引入新的工具和方法，以调查影响力病毒感染的空间组织和动态。