Functions of TBK1 in neutrophils during bacterial pneumonia

细菌性肺炎期间中性粒细胞中TBK1的功能

• 批准号: 10316200
• 负责人: Robert Stewart Hagan
• 金额: $ 7.78万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-15 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10316200
• 关键词: Anti-Bacterial Agents; Antibiotics; Autophagocytosis; Award; Bacteria; Bacterial Infections; Bacterial Pneumonia; Biology; Cell Lineage; Cells; Code; Complex; Data; Defect; Development; Equilibrium; Family; Flow Cytometry; Gene Expression; Gene Expression Profiling; Generations; Genes; Genetic Transcription; Glycolysis; Goals; Host Defense; Image; Immune; Immune response; Immune system; Impairment; Infection; Infection Control; Inflammatory; Injury; Interferons; Knockout Mice; Leukocytes; Longevity; Lung; Lung infections; Mediator of activation protein; Messenger RNA; Metabolic Pathway; Metabolism; Methods; Microscopy; Mitochondria; Molecular; Morbidity - disease rate; Mus; Myelogenous; Myeloid Cells; NADP; NADPH Oxidase; Natural Immunity; Organ; Output; Oxidases; Pathway Analysis; Pathway interactions; Patient-Focused Outcomes; Peptide Hydrolases; Persons; Phenotype; Phosphotransferases; Pneumonia; Production; Proteins; Publications; Pulmonary Inflammation; Quantitative Reverse Transcriptase PCR; Reactive Oxygen Species; Research; Resolution; Signal Transduction; Site; Speed; Streptococcus pneumoniae; TANK-binding kinase 1; Technology; Testing; Therapeutic; Tissues; Transcriptional Activation; Viral; Virus Diseases; cell motility; chemokine; cytokine; glucose uptake; healing; improved; in vivo; influenza infection; innate immune function; lung injury; macrophage; monocyte; mortality; neutrophil; novel; pathogen; pathogenic bacteria; pathogenic virus; prevent; programs; receptor; response; targeted treatment; transcription factor; transcriptome sequencing; transcriptomics

Project Summary Bacterial pneumonia causes significant morbidity and mortality worldwide despite the availability of antibiotics. The immune system must balance the competing requirements of clearing the pathogen while avoiding local organ damage. There remains an unmet need for host-directed therapeutics that limit injury or promote resolution without impairing pathogen control. Neutrophils constitute the initial response to bacterial pneumonia and control infection through direct killing of bacteria and production of mediators including chemokines, cytokines, proteases and reactive oxygen species (ROS). This R03 application aims to define novel functions for the innate immune kinase TANK-binding kinase 1 (TBK1) in neutrophils during pneumonia. TBK1 promotes the expression of type 1 interferons (IFN) by phosphorylating transcription factors from the interferon regulatory family (IRF) during viral infection, but its functions in bacterial pneumonia and in neutrophils remain undefined. My K08 Award investigates how TBK1 functions in monocytes and macrophages during influenza infection. Data from that project and from our previous publication show that during S. pneumoniae pneumonia, lung neutrophils upregulate genes coding for TBK1 and its accessory proteins, but fail to express the common downstream product, type 1 IFN, suggesting that neutrophils use the TBK1 signaling module in a unique way. We find that in S. pneumoniae pneumonia, TBK1 is required for multiple aspects of neutrophil function, including elaboration of multiple cytokines, production of reactive oxygen species (ROS), and bacterial clearance. We hypothesize that neutrophil TBK1 regulates both IRF-dependent and IRF-independent transcriptional programs as well as other cellular programs such as metabolism. The proposed aims focus on identifying cellular and molecular details of TBK1 signaling in neutrophils in response to infection by S. pneumoniae. In Aim 1, we will determine the transcriptional programs driven by TBK1 in lung neutrophils, characterizing in particular 1) the activation of transcription factors by assessing the expression of their target genes and 2) the expression of genes that regulate metabolic processes. In Aim 2, we will determine how TBK1 promotes neutrophil ROS production, with a focus on NADPH oxidase assembly and neutrophil metabolism. We will assess glucose uptake, autophagy, and mitochondrial function in lung neutrophils from WT and TBK1 KO mice in the setting of pneumonia. The proposed studies will explore new functions for the kinase TBK1 with the ultimate goal of improving patient outcomes in this common form of lung injury. These studies diverge from my K08 Award through their focus on neutrophils and bacterial pneumonia and their need for technologies unfamiliar to me. They will increase the breadth and depth of my research program to engage both myeloid cell lineages and both viral and bacterial pathogens.

项目摘要 尽管抗生素的可用性，细菌性肺炎仍在全球范围内引起显著的发病率和死亡率。 免疫系统必须平衡清除病原体的竞争需求，同时避免局部感染。 器官损伤对于限制损伤或促进免疫应答的宿主定向治疗剂仍然存在未满足的需求。 解决方案，而不损害病原体控制。中性粒细胞是细菌性肺炎的初始反应 并通过直接杀死细菌和产生包括趋化因子的介质来控制感染， 细胞因子、蛋白酶和活性氧（ROS）。此R03应用程序旨在定义新功能 对于肺炎期间中性粒细胞中的先天免疫激酶TANK结合激酶1（TBK1）。TBK1促进 1型干扰素（IFN）的表达通过磷酸化转录因子从干扰素调节蛋白， 在病毒感染过程中，它与IRF家族（IRF）的功能密切相关，但在细菌性肺炎和中性粒细胞中的功能尚不清楚。 我的K08奖研究了流感感染期间TBK1如何在单核细胞和巨噬细胞中发挥作用。 从该项目和我们以前的出版物的数据表明，在S。肺炎，肺 中性粒细胞上调编码TBK1及其辅助蛋白的基因，但不能表达常见的 下游产物，1型IFN，表明中性粒细胞以独特的方式使用TBK1信号传导模块。 我们发现，在S。肺炎，TBK1是中性粒细胞功能的多个方面所必需的， 包括多种细胞因子的产生、活性氧（ROS）的产生和细菌的产生。 间隙我们假设中性粒细胞TBK1调节IRF依赖性和IRF非依赖性 转录程序以及其他细胞程序如代谢。拟议目标 重点是通过以下方法确定中性粒细胞对感染的反应中TBK1信号传导的细胞和分子细节 S.肺炎。在目标1中，我们将确定肺中性粒细胞中TBK1驱动的转录程序， 具体地，表征1）通过评估转录因子的靶标的表达来活化转录因子， 基因和2）调节代谢过程的基因的表达。在目标2中，我们将确定如何 TBK1促进中性粒细胞ROS产生，重点是NADPH氧化酶组装和中性粒细胞 新陈代谢.我们将评估肺中性粒细胞的葡萄糖摄取、自噬和线粒体功能， 肺炎背景下的WT和TBK1 KO小鼠。拟议的研究将探讨 激酶TBK1，最终目标是改善这种常见形式的肺损伤的患者结局。这些 研究与我的K08奖不同，因为他们关注中性粒细胞和细菌性肺炎， 对于我不熟悉的技术。他们将增加我的研究计划的广度和深度， 骨髓细胞谱系以及病毒和细菌病原体。