The immunometabolite itaconate drives S. aureus lung infection

免疫代谢物衣康酸驱动金黄色葡萄球菌肺部感染

基本信息

批准号：
10383988
负责人：
Kira Leigh Tomlinson
金额：
$ 4.96万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-01-01 至 2026-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10383988
关键词：
Address Anti-Inflammatory Agents Antibiotic susceptibility Antibiotics Antiinflammatory Effect Automobile Driving Bacteria Biological Assay Bronchoalveolar Lavage Fluid Carbon Cells Chronic Clinical Communities Data Elements Energy Metabolism Environment Enzymes Equilibrium Exhibits Exposure to Flow Cytometry Future Genes Genetic Transcription Genus staphylococcus Glycolysis Goals Growth Immune Immune response In Vitro Infection Inflammation Inflammatory Lung Lung diseases Lung infections Mediating Mediator of activation protein Metabolic Metabolic stress Metabolism Microbial Biofilms Modeling Monitor Mus Nosocomial pneumonia Pathogenesis Pathway interactions Patient Isolators Patients Phagocytes Phagocytosis Phenotype Play Pneumonia Population Production Quantitative Reverse Transcriptase PCR Reporting Respiratory Burst Role Signal Transduction Source Staphylococcal Pneumonia Staphylococcus aureus Staphylococcus aureus infection Stimulus Structure of parenchyma of lung Testing Therapeutic Tissues Training Western Blotting antibiotic tolerance antimicrobial bacterial metabolism biological adaptation to stress cell type clinically relevant cytokine differential expression immunoregulation in vivo macrophage metabolomics mortality risk neutrophil pathogen pathogenic bacteria pulmonary function recruit response single-cell RNA sequencing synthetic enzyme

项目摘要

PROJECT SUMMARY/ABSTRACT Staphylococcus aureus causes community-acquired and healthcare-associated pneumonias that drive a decline in lung function and an increased risk of mortality, especially in patients with preexisting lung disease. These infections are difficult to eradicate because S. aureus can transition to adaptive phenotypes like persister cells and biofilm, which protect the bacteria against host phagocytes and antimicrobial factors, as well as antibiotics. A key component of the host immune response to pathogens is the production of regulatory metabolites like itaconate, which is synthesized in immune cells by the enzyme Immune-Responsive Gene 1 (Irg1). Itaconate balances pro- and anti-inflammatory signaling in host immune cells and exerts metabolic stress on bacteria. We recently demonstrated that itaconate inhibits S. aureus glycolysis and restructures staphylococcal metabolism to promote biofilm formation. Itaconate also limited the use of key energy-producing pathways, suggesting that it may promote the formation of antibiotic-tolerant persister cells. The role of itaconate in driving S. aureus lung infections cannot be completely understood without also investigating its impact on the host response to S. aureus. Preliminary data demonstrate that Irg1 is highly expressed in neutrophils and is associated with increased pro-inflammatory cytokine production during S. aureus lung infection, which differs from the anti- inflammatory effects of itaconate that have been defined in other cell types and infection models. This proposal aims to further investigate itaconate as a central mediator of the host-pathogen dynamic during S. aureus lung infection and a potential driver of S. aureus adaptation to the lung. Aim 1 will define the role of itaconate in regulating the host immune response to S. aureus by using metabolomics, phagocytosis assays, oxidative burst assays, and scRNA-sequencing to 1) determine if neutrophils are a major source of itaconate during S. aureus infection, 2) define the impact of itaconate on neutrophil effector function, and 3) identify the pathways that are differentially regulated by itaconate in neutrophils and other immune cell populations during in vivo infection. Aim 2 will define the role of itaconate in driving S. aureus adaptation to the lung by using bacterial qRT-PCR, ATP quantification, antibiotic tolerance assays, and flow cytometry to 1) determine if itaconate drives transcriptional changes that promote persister cell formation in vivo, 2) establish that itaconate exposure drives reduced energy metabolism and increased antibiotic tolerance ex vivo, and 3) quantify S. aureus division and growth in response to itaconate in vivo. Together, these studies will define the role of host immunometabolism in driving inflammation and bacterial persistence during S. aureus lung infection.

项目概要/摘要金黄色葡萄球菌引起社区获得性肺炎和医疗保健相关肺炎，导致感染率下降肺功能受损，死亡风险增加，尤其是患有肺部疾病的患者。这些感染很难根除，因为金黄色葡萄球菌可以转变为像持续细胞这样的适应性表型和生物膜，保护细菌免受宿主吞噬细胞、抗菌因子以及抗生素的侵害。宿主对病原体的免疫反应的一个关键组成部分是调节代谢物的产生，例如衣康酸，由免疫反应基因 1 (Irg1) 在免疫细胞中合成。衣康酸平衡宿主免疫细胞中的促炎和抗炎信号传导，并对细菌施加代谢压力。我们最近证明衣康酸抑制金黄色葡萄球菌糖酵解并重组葡萄球菌代谢以促进生物膜的形成。衣康酸还限制了关键能量产生途径的使用，这表明它可能促进抗生素耐受性持续细胞的形成。衣康酸在驱动金黄色葡萄球菌肺中的作用如果不研究其对宿主对金黄色葡萄球菌反应的影响，就无法完全了解感染。金黄色葡萄球菌。初步数据表明，Irg1 在中性粒细胞中高表达，并与金黄色葡萄球菌肺部感染期间促炎细胞因子的产生增加，这与抗炎细胞因子不同衣康酸的炎症作用已在其他细胞类型和感染模型中得到定义。该提案旨在进一步研究衣康酸作为宿主-病原体动态的中心调节剂金黄色葡萄球菌肺部感染期间和金黄色葡萄球菌适应肺部的潜在驱动因素。目标 1 将定义利用代谢组学、吞噬作用研究衣康酸在调节宿主对金黄色葡萄球菌免疫反应中的作用测定、氧化爆发测定和 scRNA 测序，以 1) 确定中性粒细胞是否是主要来源金黄色葡萄球菌感染期间的衣康酸，2) 定义衣康酸对中性粒细胞效应功能的影响，以及 3) 确定中性粒细胞和其他免疫细胞中衣康酸差异调节的途径体内感染期间的人群。目标 2 将定义衣康酸在推动金黄色葡萄球菌适应环境中的作用通过使用细菌 qRT-PCR、ATP 定量、抗生素耐受性测定和流式细胞术来检测肺部 1) 确定衣康酸是否驱动促进体内持久细胞形成的转录变化，2) 建立衣康酸暴露会导致能量代谢降低和离体抗生素耐受性增加，以及 3) 量化体内衣康酸对金黄色葡萄球菌的分裂和生长的反应。这些研究将共同定义金黄色葡萄球菌肺部感染期间宿主免疫代谢在驱动炎症和细菌持续存在中的作用。