MECHANISTIC UNDERSTANDING OF INFLUENZA-HOST INTERACTIONS FROM A ZOONOTIC PERSPECTIVE

从人畜共患病的角度理解流感-宿主相互作用的机制

• 批准号: 10217310
• 负责人: Nicholas C. Wu
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-21 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10217310
• 关键词: Affinity; Amino Acid Sequence; Amino Acid Substitution; Antiviral Agents; Avian Influenza A Virus; Binding; Biochemistry; Biological; Biophysical Process; Birds; Cells; Coiled-Coil Domain; Complex; Coupling; Cryoelectron Microscopy; Crystallization; Data; Disease Outbreaks; Engineering; Epidemic; Genetic; Genetic Variation; Genome; Goals; H7N7; Hemagglutinin; Human; Immune; Immune Evasion; Individual; Influenza; Influenza A Virus, H1N1 Subtype; Influenza A Virus, H5N1 Subtype; Influenza A Virus, H7N9 Subtype; Influenza A virus; Innate Immune Response; Integration Host Factors; Interferons; Intervention; Length; Maps; Mass Spectrum Analysis; Measures; Mediating; Membrane Glycoproteins; Molecular; Molecular Biology; Mutagenesis; Mutation; Neuraminidase; Outcome; Pathogenicity; Phase; Play; Population; Property; Proteins; Publishing; RNA Viruses; Recombinant Proteins; Research; Risk; Role; Species Specificity; Specificity; Structure; Systems Biology; TRIM25 gene; Technology; Tropism; Validation; Variant; Viral Proteins; Virus; Yeasts; Zoonoses; aquatic bird; base; biophysical properties; cross-species transmission; fitness; influenzavirus; insight; mortality; mutation screening; next generation sequencing; novel; pandemic disease; particle; preference; prevent; protein protein interaction; receptor; seasonal influenza; socioeconomics; structural biology; transmission process; ubiquitin-protein ligase; virus host interaction

PROJECT SUMMARY Influenza A virus is a negative-strand RNA virus and is classified into subtypes based on the antigenic properties of the two surface glycoproteins, namely hemagglutinin (HA) and neuraminidase (NA). There are 18 known HA subtypes (H1 to H18) and 11 known NA subtypes (N1 to N11). Wild aquatic birds are the main natural reservoir for influenza A viruses. However, spillover from natural reservoirs can cause human pandemics. Three subtypes (H1N1, H2N2, and H3N2) have caused human pandemics, and two of them (H1N1 and H3N2) are presently circulating the in human population, causing seasonal outbreaks. Other subtypes found in natural reservoirs also occasionally transmit to the human population, as exemplified by H5N1, H5N6, H6N1, H7N7, H7N9, H9N2, and H10N8 viruses. These zoonotic subtypes can be highly pathogenic and have a high mortality rate (>50% of hospitalized individuals) when infecting humans. The virus- host protein-protein interaction constitutes the molecular basis of influenza pathogenicity. Many “omics” studies have explored influenza-host interactions. However, the known mechanistic details are far from comprehensive, especially regarding variation in pathogenicity across different virus subtypes and hosts. The objective of the proposed study is to understand how genetic variation in influenza virus and the host together influence pathogenicity. This proposal will focus on influenza NS1 protein, which is a major pathogenicity determinant and plays a role in host adaptation. NS1 is known to interact with many host proteins. Some of these interactions are known to be influenza subtype-dependent. In the proposed study, NS1 from two seasonal subtypes (H1N1 and H3N2) and one zoonotic subtype (H5N1) will be employed. The difference in the functional constraints of these three NS1 in human and avian cells will be interrogated by deep mutational scanning and systematic interactome analysis (K99 phase). Subsequently, biophysical characterization of NS1-TRIM25 interaction will be performed. NS1 interacts with TRIM25, which is an E3 ubiquitin ligase, to suppress the innate immune response. NS1-TRIM25 interaction is virus subtype-dependent and host-dependent. The underlying biophysical mechanism will be probed by affinity maturation and cryo- electron microscopy (K99/R00 phase). The proposed study will facilitate a detailed understanding of influenza pathogenicity and tropism from the molecular biology, genetics, evolutionary, and structural biology perspectives. More importantly, the integrative research framework developed in this study will be applicable to other influenza virus proteins, or even to proteins from other viruses.

项目概要 甲型流感病毒是一种负链RNA病毒，根据抗原分为亚型 两种表面糖蛋白的特性，即血凝素（HA）和神经氨酸酶（NA）共有18种。 已知的HA亚型（H1至H18）和11种已知的NA亚型（N1至N11）是主要的野生水禽。 甲型流感病毒的天然宿主 然而，天然宿主的溢出可能会导致人类感染。 三种亚型（H1N1、H2N2 和 H3N2）引起了人类大流行，其中两种亚型。 （H1N1 和 H3N2）目前在人群中传播，导致季节性爆发。 在天然宿主中发现的亚型有时也会传播给人类，例如 H5N1、H5N6、H6N1、H7N7、H7N9、H9N2 和 H10N8 病毒可能是高度人畜共患病的亚型。 具有致病性，感染人类时死亡率很高（>50% 的住院患者）。 宿主蛋白质-蛋白质相互作用构成了流感致病性的分子基础 许多“组学”研究。 已经探索了流感与宿主的相互作用，但是，已知的机制细节还很遥远。 全面，特别是关于不同病毒亚型和宿主的致病性差异。 拟议研究的目的是了解流感病毒的遗传变异和 该提案将重点关注流感 NS1 蛋白，这是一个主要的蛋白。 NS1 是致病性决定因素并在宿主适应中发挥作用，已知与许多宿主相互作用。 在拟议的研究中，已知其中一些相互作用与流感亚型有关。 将使用来自两种季节性亚型（H1N1 和 H3N2）和一种人畜共患亚型（H5N1）的 NS1。 人类和鸟类细胞中这三种 NS1 的功能限制的差异将通过 深度突变扫描和系统相互作用组分析（K99 阶段）。 将进行 NS1-TRIM25 相互作用的表征，NS1 与 TRIM25 相互作用，TRIM25 是 E3。 泛素连接酶，抑制先天免疫反应的 NS1-TRIM25 相互作用是病毒亚型依赖性的。 潜在的生物物理机制将通过亲和力成熟和冷冻来探索。 电子显微镜（K99/R00 相）。拟议的研究将有助于详细了解流感。 分子生物学、遗传学、进化论和结构生物学的致病性和趋向性 更重要的是，本研究开发的综合研究框架将适用于 其他流感病毒蛋白质，甚至其他病毒的蛋白质。