Macrophage Metabolism After Target Cell Ingestion Regulates Anti-Inflammatory Reprogramming

靶细胞摄入后巨噬细胞代谢调节抗炎重编程

• 批准号: 10239261
• 负责人: Alexandra Leigh McCubbrey
• 金额: $ 24.9万
• 依托单位: NATIONAL JEWISH HEALTH
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-15 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10239261
• 关键词: Acute; Address; Advisory Committees; Affect; Amino Acids; Anti-Inflammatory Agents; Apoptotic; Arginine; Biology; Bone Marrow; Cells; Chimera organism; Data; Development; Dose; Enzymes; Evaluation; Failure; Foundations; Genetic Transcription; Human; Immune; In Vitro; Inflammasome; Inflammation; Inflammatory; Ingestion; Injury; Interleukin-1 beta; Investigation; Isotopes; Journals; Knowledge; Laboratories; Learning; Leukocytes; Lung Inflammation; Lung diseases; Measures; Mediating; Mentors; Metabolic; Metabolism; Molecular Target; Mus; Necrosis; Pharmacology; Phase; Polyamines; Process; Production; Putrescine; Reporting; Research; Resolution; Role; Scientist; Signal Transduction; Source; Spermidine; Spermine; System; Techniques; Testing; Transgenic Organisms; Up-Regulation; Work; arginase; congenic; cytokine; experience; histone methylation; histone modification; in vivo; inhibitor/antagonist; lung injury; lung repair; macromolecule; macrophage; metabolomics; monocyte; prevent; pulmonary function; recruit; response; restoration; symposium; transcriptome sequencing

ABSTRACT Inflammation is a ubiquitous component of lung disease involving accumulation of leukocytes in the airspaces. In order for inflammation to resolve, dead and dying leukocytes must be removed and production of inflammatory cytokines must be turned off. Macrophages (Mϕ) are key orchestrators of these processes, however the triggers that reprogram inflammatory Mϕ to perform these roles remain incompletely understood. Clearance of dead cells has been shown to provide an important reprogramming signal, but we have limited knowledge of the precise mechanisms by which dead cell ingestion facilitates redirection of Mϕ function. Recent work has demonstrated a key role for cellular metabolism in determining Mϕ function. Ingested target cells provide a clear source of varied macromolecules that must be digested by Mϕ. However, little research has been done to assess intracellular metabolites produced by this process or to consider the subsequent immune consequence. We propose the first comprehensive study of Mϕ metabolism following target cell ingestion and degradation, and hypothesize that changes in the levels of intracellular metabolites control ingestion-driven Mϕ reprogramming. Our preliminary studies have identified a promising molecular target, polyamines, which are dramatically increased in Mϕ following the ingestion of apoptotic target cells. Polyamines have a known anti- inflammatory function including suppression of numerous pro-inflammatory cytokines. We propose to test the hypothesis that upregulation of polyamine synthesis by Mϕ following target cell ingestion is critical to suppress a key Mϕ cytokine IL-1β, and important for the resolution of lung inflammation in vivo. During the K99 mentored phase, I will build upon my experience studying Mϕ biology and target cell clearance by learning techniques related to cellular metabolism including unbiased metabolomics and the use of isotope metabolites to measure metabolic flux. During this phase, I will: 1) complete a comprehensive, integrated assessment of the metabolites produced following ingestion of target cells, focusing on polyamines, and assess whether these metabolites derive from digested target cell material, and 2) specifically assess the role of arginase-1 in regulating polyamine synthesis by inflammatory Mϕ. Simultaneously, I will enrich my professional development by participating in journal clubs, seminars, national conferences, coursework, and having semi-annual evaluations by a trainee advisory committee. The R00 independent phase will allow me to establish my laboratory as I continue investigation into: 3) how Mϕ polyamines affect cytokine production after target cell ingestion, and 4) the effects of Mϕ polyamines on resolution of inflammation and lung repair. Collectively, this proposal will enhance my current expertise, address a critical unknown in the field of target cell clearance, and provide the necessary foundation to establish myself as an independent research scientist.

抽象的 炎症是肺部疾病普遍存在的一个组成部分，涉及白细胞在肺部的积聚。 空域。为了消除炎症，必须清除死亡和垂死的白细胞并产生 必须关闭炎症细胞因子。巨噬细胞 (Mphi) 是这些过程的关键协调者， 然而，重新编程炎症 M 来发挥这些作用的触发因素仍然不完全清楚。 死亡细胞的清除已被证明可以提供重要的重编程信号，但我们的研究有限 了解死细胞摄入促进 Mψ 函数重定向的精确机制。 最近的工作证明了细胞代谢在确定 M 函数中的关键作用。摄取目标 细胞提供了各种大分子的明确来源，这些大分子必须被 Mφ 消化。但研究甚少 已完成评估该过程产生的细胞内代谢物或考虑后续 免疫后果。 我们提出了第一个对靶细胞摄入后 Mφ 代谢的综合研究， 降解，并假设细胞内代谢物水平的变化控制摄入驱动的 Mφ 重新编程。我们的初步研究已经确定了一个有前途的分子靶点——多胺，它是 摄入凋亡靶细胞后 M 显着增加。多胺具有已知的抗 炎症功能，包括抑制多种促炎细胞因子。我们建议测试 假设靶细胞摄入后 Mphi 上调多胺合成对于抑制 一种关键的 Mphi 细胞因子 IL-1β，对于体内肺部炎症的解决很重要。 在 K99 指导阶段，我将利用我研究 Mφ 生物学和靶细胞的经验 通过学习与细胞代谢相关的技术（包括无偏见的代谢组学和使用）来清除 同位素代谢物来测量代谢通量。在此阶段，我将：1）完成全面的、 对摄入靶细胞后产生的代谢物进行综合评估，重点是多胺， 并评估这些代谢物是否源自消化的靶细胞材料，以及 2) 特别评估 arginase-1 在通过炎症 Mphi 调节多胺合成中的作用。同时，我也会丰富自己的 通过参加期刊俱乐部、研讨会、全国会议、课程作业和 实习生咨询委员会每半年进行一次评估。 R00 独立阶段将允许我 建立我的实验室，同时我继续研究：3）M phi 多胺如何影响细胞因子的产生 靶细胞摄入，以及 4) M 多胺对炎症消退和肺部修复的影响。 总的来说，该提案将增强我目前的专业知识，解决目标领域的一个关键未知问题 细胞清除，并为自己成为一名独立研究科学家提供必要的基础。