The role of MIF in mitochondrial metabolism and M2-TAM polarization

MIF 在线粒体代谢和 M2-TAM 极化中的作用

• 批准号: 9759348
• 负责人: Jordan T Noe
• 金额: $ 3.16万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-01 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9759348
• 关键词: Adopted; Anaerobic Bacteria; Binding; Biogenesis; Biological Assay; COPS5 gene; Cancer Patient; Citric Acid Cycle; Clinical; Complex; Data; Development; Diagnosis; Electron Transport; Exhibits; Flow Cytometry; Fumarates; GA-binding protein transcription factor; Gene Expression Profiling; Genetic; Hypoxia Inducible Factor; IL4 gene; Immunosuppression; Immunosuppressive Agents; Immunotherapeutic agent; Immunotherapy; Incidence; Interleukin-4; Laboratories; Lead; Link; Malignant Neoplasms; Mediating; Metabolic; Metabolic Pathway; Metabolism; Migration Inhibitory Factor; Mitochondria; Modeling; Molecular Target; Mus; Neoplasm Metastasis; Oxidative Phosphorylation; Oxides; Oxygen Consumption; Pathway interactions; Patients; Phenotype; Process; Production; Proteins; Pyruvate; Pyruvate Metabolism Pathway; Reporter; Research; Resistance; Role; Solid; Solid Neoplasm; Stimulus; Stromal Neoplasm; Succinate Dehydrogenase; Survival Rate; Testing; Transgenic Mice; Tumor Immunity; Tumor-Derived; Tumor-associated macrophages; Unresectable; Warburg Effect; adenoviral-mediated; angiogenesis; anti-tumor immune response; cancer cell; cancer type; cytokine; density; expression vector; extracellular; gene product; glucose metabolism; improved; macrophage; melanoma; metabolic phenotype; metabolomics; mitochondrial metabolism; mortality; mutant; neoplasm immunotherapy; novel therapeutics; response; small molecule; transcription factor; tumor; tumor microenvironment; tumor progression; tumorigenic; uptake

PROJECT SUMMARY/ABSTRACT “The role of MIF in mitochondrial metabolism and M2-TAM polarization.” Tumor-associated macrophages (TAMs) promote intratumoral microvessel density, increase tumor stromal matrix remodeling and potently suppress anti-tumor immunity in late-stage cancer patients. Although substantial research has identified how “M2” TAM polarization promotes tumor progression, much less is known about how TAMs obtain this M2 phenotype within tumor microenvironments. Metabolic reprogramming is a critical determinant of macrophage polarization as recent studies demonstrate that M2 TAMs adopt an oxidative phosphorylation/mitochondrial metabolic phenotype and uniquely respond to metabolites within the tumor microenvironment. This proposal’s long-term objective is to investigate the mechanisms by which macrophage migration inhibitory factor (MIF) dictates lactate-enhanced M2 polarization. Our preliminary studies demonstrate that M2 macrophages metabolize extracellular lactate to pyruvate which, in turn, is oxidized in the mitochondria and is both necessary and sufficient to significantly enhancing Th2 cytokine-dependent M2 TAM polarization. Our data further indicates that MIF promotes mitochondrial metabolism which is required for maximal lactate/pyruvate-enhanced M2 polarization and acquisition of pro-tumorigenic phenotypes. Two aims are proposed to test our hypotheses: Specific Aim 1: Delineate the mechanistic effectors of lactate/pyruvate to HIF1α-dependent M2-macrophage polarization. Objective: To use a combination of metabolic flux analyses, metabolomics, gene expression assays, transgenic mice and mechanistic studies to answer three primary questions: 1) Whether M2 macrophages actively metabolize lactate; 2) Which metabolic pathways are utilized to accomplish this, and; 3) How is lactate metabolism mechanistically linked to M2 polarization Specific Aim 2: Determine the mechanistic contributions of MIF in lactate/pyruvate-mediated M2 TAM polarization. Objective: Use mutant MIF protein re-expression, small-molecule rescue studies, flow cytometry and gene expression assays to make three determinations: 1) Whether MIF dictates mitochondrial biogenesis and metabolism in M2 macrophages; 2) Whether and how MIF influence mitochondrial metabolism by binding to CSN5 and regulating NRF2 stability, and; 3) If MIF is functionally linked to lactate-amplified HIF1α stability and ensuing M2 polarization.

项目摘要/摘要 MIF在线粒体代谢和M2-极化中的作用。 肿瘤相关巨噬细胞(TAMs)促进肿瘤内微血管密度，增加肿瘤 间质基质重塑和有效抑制晚期癌症患者的抗肿瘤免疫。虽然 大量研究已经确定了极化是如何促进肿瘤进展的，但知之甚少 关于TAMS如何在肿瘤微环境中获得这种M2表型。代谢重新编程是一种 巨噬细胞极化的关键决定因素最近的研究表明M2 TAMs采用氧化 磷酸化/线粒体代谢表型和对肿瘤内代谢物的独特反应 微环境。这项提案的长期目标是研究巨噬细胞 迁移抑制因子(MIF)决定乳酸增强的M2极化。我们的初步研究表明 M2巨噬细胞将细胞外乳酸代谢成丙酮酸，丙酮酸在线粒体中被氧化 这是显著增强Th2型细胞因子依赖的M2极化的必要条件和充分条件。 我们的数据进一步表明，MIF促进线粒体代谢，这是最大限度地 乳酸盐/丙酮酸增强的M2极化和促肿瘤表型的获得。两个目标是 建议检验我们的假设：具体目标1：描述乳酸/丙酮酸的机械效应 HIF1α依赖的M2-巨噬细胞极化。目的：结合代谢流量分析， 代谢组学、基因表达分析、转基因小鼠和机制研究回答三个主要问题 问题：1)M2巨噬细胞是否主动代谢乳酸；2)利用哪些代谢途径 要做到这一点，以及；3)乳酸盐代谢如何机械地与M2极化特定目标联系在一起2： 确定MIF在乳酸/丙酮酸介导的M2极化中的作用机制。 目的：利用突变型MIF蛋白再表达、小分子挽救研究、流式细胞术和基因分析 表达分析做出三个决定：1)MIF是否决定线粒体的生物发生和 M2巨噬细胞的代谢；2)MIF是否以及如何通过结合 Csn5和调节NRF2的稳定性；以及；3)如果MIF在功能上与乳酸扩增的HIF1α的稳定性和 随之而来的M2极化。