Engineering host-pathogen interactions to understand influenza A infection

工程宿主-病原体相互作用以了解甲型流感感染

• 批准号: 2129624
• 负责人: Gregg Duncan
• 金额: $ 53.62万
• 依托单位: University of Maryland, College Park
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-10-01 至 2025-09-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2129624&HistoricalAwards=false
• 关键词: Engineering; host; pathogen; interactions; understand; Engineering; host; pathogen; interactions; understand

Respiratory viruses like influenza have evolved to circumvent the body’s natural defenses to infection. Following inhalation into the lung, viral particles must navigate through mucus that coats and protects that lung. However, it is not yet fully understood what allows these viruses to bypass airway mucus to reach underlying cells. This project seeks to understand how the physical size and shape of influenza virus influences its ability to overcome the mucus barrier. In addition, studies will be designed to determine how the preference of these viruses to bind to certain sugars found in abundance within mucus and on airway epithelial cells impacts their movement within mucus. This work will lay the foundation for future studies on other respiratory pathogens such as rhinovirus, respiratory syncytial virus (RSV), and coronaviruses (e.g., SARS-CoV-2). The project will provide multidisciplinary training opportunities for graduate and undergraduate students who will be actively recruited from diverse backgrounds. The goal of this project is to understand both the viral and host factors that influence the ability of Influenza A virus (IAV) to penetrate the mucosal barrier to infection. Mucus is composed of heavily glycosylated mucin proteins and presents both a biochemical and physical barrier to pathogens within the lung microenvironment. To initiate infection, influenza A virus (IAV) binds to alpha 2,3- and/or 2,6-linked sialic acid on the airway epithelial surface with a strain-dependent receptor preference. The preference of IAV for sialic acid glycoforms may alter virus interactions with mucin-associated sialic acid. Yet, it is unknown how this may influence trapping of IAV by mucus. Furthermore, IAV naturally produces virions with both spherical and filamentous shape. While other virion-associated factors in IAV infection have been explored, the role of IAV morphology remains unclear. On the host side, barrier function of mucus and its impact on IAV infection are not understood due to the challenges in effectively modeling mucosal antiviral defense. To examine these complex host-pathogen interactions in detail, the project will measure diffusion of IAV particles in mucus collected from human bronchial epithelial cultures and bioengineered synthetic mucus using fluorescent video microscopy and multiple particle tracking image analysis. The approach taken will incorporate molecular virology to produce IAV particles with specified receptor preference and shape as well as biochemical engineering methods to modulate glycan expression in synthetic mucus barriers. This comprehensive toolbox will be used to analyze the hypothesis that IAV penetration is a function of virion morphology and glycan-binding preferences as well as the composition and architecture of the mucus barrier. The unique ability to quantitatively examine IAV-mucus interactions makes this approach highly valuable and complementary to standard assays of infectivity. Building on this fundamental work, a novel high-throughput viral infectivity screening system that accounts for the impact of mucus barrier properties will be created. The unique, highly innovative tools developed through this work will enhance our understanding of IAV and offer a new perspective on the factors that influence how viruses pass through mucus prior to infection.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

呼吸道病如影响的呼吸道已经发展为将人体的自然防御能力绕开。吸入肺部后，病毒颗粒必须穿过涂层和保护肺的粘液。但是，尚未完全了解是什么允许这些病毒绕过气道粘液到达潜在的细胞。该项目试图了解病毒的身体大小和影响如何影响其克服粘液屏障的能力。此外，将设计研究以确定这些病毒在粘液中和气道上皮细胞中与某些糖结合的偏好如何影响其在粘液中的运动。这项工作将奠定对其他呼吸道病原体（例如鼻病毒，呼吸道合胞病毒（RSV）和冠状病毒（例如SARS-COV-2））的未来研究的基础。该项目将为研究生和本科生提供多学科的培训机会，他们将积极从潜水员背景中招募。该项目的目的是了解影响流感病毒（IAV）渗透粘膜屏障感染能力的病毒和宿主因素。粘液由大量糖基化的粘蛋白蛋白组成，并为肺微环境内病原体的生化和物理障碍提供了生化和物理障碍。为了引发感染，影响病毒（IAV）与α2,3-和/或2,6连锁的唾液酸在气道上皮表面上具有菌株依赖性受体偏好。 IAV对唾液酸糖型的偏爱可能会改变与肉豆素相关的唾液酸的病毒相互作用。然而，尚不清楚这可能会影响mutus捕获IAV。此外，IAV自然会产生带球形和丝状形状的病毒。虽然已经探索了IAV感染中其他与病毒体相关的因素，但IAV形态的作用尚不清楚。在宿主方面，由于有效建模粘膜抗病毒防御的挑战，粘液的屏障功能及其对IAV感染的影响尚不清楚。为了详细检查这些复杂的宿主 - 病原体相互作用，该项目将使用荧光视频显微镜和多个粒子跟踪图像分析来测量从人支气管上皮培养物中收集的粘液中IAV颗粒的扩散和生物工程的合成mutus。采用的方法将结合分子病毒学，以产生具有指定受体偏好和形状以及生化工程方法的IAV颗粒，以调节合成mutus屏障中的聚糖表达。该全面的工具箱将用于分析以下假设：IAV渗透是病毒体形态和聚糖结合偏好的函数以及MUTUS屏障的组成和结构。定量检查IAV-MUCUS相互作用的独特能力使得这种方法具有高价值和与标准感染的互补性。在这项基本工作的基础上，将创建一种新型的高通量病毒感染筛查系统，该系统将创建粘液屏障特性的影响。通过这项工作开发的独特，高度创新的工具将增强我们对IAV的理解，并为影响病毒在感染前通过粘液通过粘液的因素提供了新的观点。该奖项反映了NSF的法定任务，并通过使用基金会的知识分子和更广泛的影响审查标准来通过评估来诚实地通过评估来诚实地支持。