The Role of TRIM6 and Ubiquitin in Influenza Virus-Induced Pathology

TRIM6 和泛素在流感病毒诱发的病理学中的作用

基本信息

批准号：
10596915
负责人：
Ricardo Rajsbaum
金额：
$ 58.24万
依托单位：
RBHS-NEW JERSEY MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-05-13 至 2023-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10596915
关键词：
Affect Anti-Inflammatory Agents Antiviral Response Biological Testing Bone Marrow CXCL1 gene Cell membrane Cells Cessation of life Chemotactic Factors Chimera organism Co-Immunoprecipitations Complex Coronavirus Cytokine Signaling Dangerousness Data Development Disease Epidemic Equilibrium Flow Cytometry Future Goals IL8 gene Immune Impairment Infection Inflammation Inflammatory Influenza A virus Interferon Type I Interferons Intervention Knockout Mice Knowledge Learning Lung Mediating Molecular Morbidity - disease rate Mus Mutation Neutrophil Infiltration Outcome PI3K/AKT Pathogenicity Pathology Pathway interactions Pharmacology Phosphorylation Phosphorylation Site Phosphotransferases Play Polyubiquitin Production Proteins Proto-Oncogene Proteins c-akt Pulmonary Inflammation Regulation Reporting Role Signal Induction Signal Transduction Source System TNF gene TNFRSF1A gene Time Tissues Ubiquitin Ubiquitination Virus Virus Diseases Work antagonist arm base cell type chemokine cytokine improved in vitro Assay in vivo influenzavirus inhibitor innate immune mechanisms monocyte mortality neutrophil next generation sequencing novel novel therapeutic intervention pandemic disease public health relevance receptor recruit response single-cell RNA sequencing targeted treatment therapeutic development tool ubiquitin-protein ligase

项目摘要

ABSTRACT. Influenza A virus (IAV) causes annual epidemics and dangerous pandemics involving millions of cases of illness and deaths worldwide. The main cause of pathology from IAV is excessive inflammation, therefore, the overarching goal of this proposal is to learn how mechanisms of inflammation can be manipulated to promote disease tolerance to virus infection. Cytokine production, a chief contributor of inflammation, is regulated at the post-translational level to balance between efficient antiviral responses and damaging inflammation. A major molecular regulatory mechanism involves ubiquitination of signaling components. The specific goal of this proposal is to identify mechanisms of regulation of inflammation by the ubiquitin (Ub) system during IAV infection in vivo. We recently reported that the E3-Ub ligase, TRIM6, catalyzes the synthesis of unanchored poly-Ub chains, which promote antiviral IFN-I responses. However, the role of TRIM6 in regulating other inflammatory cytokines is not known. We generated TRIM6 knockout mice (Trim6-/-), which provides a unique tool to identify novel immune pathways regulated by TRIM6 and unanchored Ub in vivo. Our preliminary data show that Trim6-/- mice have fewer signs of pathology even though there are increased IAV titers at early time points post-infection. We also found reduced expression levels of CXCL1, a well-known neutrophil chemo-attractant, which correlated with reduced neutrophil infiltration to the lungs of IAV-infected Trim6-/- mice. We found that TRIM6 and unanchored Ub form a complex with PI3K/AKT signaling components, and their phosphorylation is impaired in Trim6-/- cells. Our data also suggest that TNFα produced by infected cells induces pathogenic CXCL1 in bystander cells to recruit neutrophils. Neutrophils are known to be recruited to the lung during IAV infection and can play both protective and detrimental roles. However, what factors drive neutrophils to cause tissue damage during infection are not well-understood. Therefore, there is a gap in knowledge on the mechanisms of regulation of neutrophil recruitment and their roles in the balance between protective responses and pathogenic inflammation. Our hypothesis is that TRIM6 is activated by TNFα signaling and promotes early CXCL1-mediated pathogenic inflammation, thereby inhibiting disease tolerance. In Aim 1, we will determine the cellular source of TRIM6-induced CXCL1, and its role in neutrophil recruitment to the lungs, during IAV infection. We will demonstrate the role of early CXCL1 production in pathology and whether TNFα is involved in inducing TRIM6-mediated CXCL1. In Aim 2, we will determine the mechanism by which TRIM6 and Ub modulate the activation of PI3K-AKT for downstream signaling and how TRIM6 is activated during infection. The outcomes include the identification of the cellular source of pathogenic CXCL1, and the mechanism by which TRIM6 is activated for signaling. This information will guide the development of therapeutic approaches by targeting TRIM6 and CXCL1-producing cells to reduce inflammatory diseases.

抽象的。流感病毒（IAV）导致年度流行病和危险大流行病，涉及数百万全球疾病和死亡的病例。 IAV病理学的主要原因是过度炎症，因此，该提案的总体目标是了解炎症机制如何操纵以促进对病毒感染的疾病耐受性。 Cytokine生产，主要贡献者炎症在翻译后水平受到调节，以平衡有效的抗病毒反应和破坏性炎症。主要的分子调节机制涉及信号传导的泛素化成分。该提案的具体目标是确定通过在体内IAV感染期间，泛素（UB）系统。我们最近报道，E3-UB连接酶TRIM6催化了未经锚定的聚-UB链的合成，促进抗病毒IFN-I反应。但是，TRIM6在控制其他炎症中的作用细胞因子尚不清楚。我们生成了TRIM6敲除小鼠（TRIM6 - / - ），该小鼠为识别由TRIM6调节的新型免疫途径和在体内未经锚定的UB。我们的初步数据显示 Trim6 - / - 小鼠的病理迹象较少，即使早期的IAV滴度增加了感染后。我们还发现CXCL1的表达水平降低，CXCL1是一种众所周知的中性粒细胞化学吸引剂，与降低的嗜中性粒细胞浸润对IAV感染的Trim6 - / - 小鼠的肺部相关。我们发现 TRIM6和未经锚定的UB形成具有PI3K/AKT信号成分的复合物，其磷酸化为在TRIM6 - / - 细胞中受损。我们的数据还表明，受感染细胞产生的TNFα诱导致病性旁观者细胞中的CXCL1募集中性粒细胞。已知中性粒细胞在IAV期间被招募到肺部感染，可以发挥受保护和有害作用。但是，哪些因素驱动中性粒细胞引起感染过程中的组织损伤并不理解。因此，知道中性粒细胞募集的调节机制及其在受保护之间的平衡中的作用反应和致病性炎症。我们的假设是TRIM6被TNFα信号传导激活，并且促进早期CXCL1介导的致病感染，从而抑制疾病的耐受性。在AIM 1中，我们将确定TRIM6诱导的CXCL1的细胞来源，及其在中性粒细胞募集到肺中的作用，在IAV感染期间。我们将证明早期CXCL1产生在病理学中的作用以及TNFα是否参与诱导的TRIM6介导的CXCL1。在AIM 2中，我们将确定修剪的机制 UB调节PI3K-AKT的激活以进行下游信号，以及如何激活TRIM6。感染。结果包括鉴定致病性CXCL1的细胞来源，以及将TRIM6激活以进行信号传导的机制。此信息将指导开发通过靶向TRIM6和CXCL1产生细胞来减少炎症性疾病，采用治疗方法。