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: 10199254
• 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/10199254
• 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; Respiratory physiology; 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; 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 炎症是肺部疾病的普遍成分，涉及白细胞在 空域。为了解决炎症，必须去除死亡和垂死的白细胞，并产生 炎症细胞因子必须关闭。巨噬细胞（Mϕ）是这些过程的关键编排， 然而，重编程炎症性m手的触发器仍未完全理解。 已显示死细胞的清除提供了重要的重编程信号，但是我们有限 了解死细胞摄取设施的确切机制的Mϕ功能。 最近的工作表明了细胞代谢在确定Mϕ功能中的关键作用。摄入的目标 细胞提供了必须通过M ϕ消化的各种大分子的清晰来源。但是，很少的研究 已经进行了评估该过程产生的细胞内代谢产物或考虑随后的 免疫后果。 我们提出了目标细胞摄入后的M ϕ代谢的首次全面研究， 降解，并假设细胞内代谢产物控制摄入驱动的M ϕ的水平的变化 重新编程。我们的初步研究已经确定了一个有希望的分子靶标，多胺是 摄入凋亡靶细胞后，Mϕ动态增加。多胺具有已知的抗 炎症功能，包括抑制多种促炎性细胞因子。我们建议测试 假设在靶细胞摄入后通过M ϕ上调多胺合成对于抑制至关重要 关键的M ϕ细胞因子IL-1β，对于在体内肺注射的分辨率很重要。 在K99修补阶段，我将基于研究M ϕ生物学和靶细胞的经验 通过与细胞代谢有关的学习技术清除，包括无偏代谢组学和使用 同位素代谢物的测量代谢通量。在此阶段，我将：1）完成全面的， 摄入靶细胞后产生的代谢产生的代谢产物的综合评估，重点是多胺， 并评估这些代谢产物是否来自消化的靶细胞材料，2）专门评估 精氨酸酶-1在通过炎症Mϕ控制多胺合成中的作用。同时，我会丰富我的 通过参加期刊俱乐部，半货人，民族会议，课程工作和专业发展 由学员咨询委员会进行半年度评估。 R00独立阶段将使我能够 在我继续投资投资时，建立我的实验室：3）M ϕ多胺如何影响细胞因子的产生 靶细胞摄入，以及4）Mϕ多胺对感染和肺修复的分辨率的影响。 总的来说，该建议将增强我当前的专业知识，解决目标领域的关键未知 细胞清除，并为建立自己的独立研究科学家提供必要的基础。