Academic Journal
Pro-inflammatory macrophages produce mitochondria-derived superoxide by reverse electron transport at complex I that regulates IL-1β release during NLRP3 inflammasome activation.
| Τίτλος: | Pro-inflammatory macrophages produce mitochondria-derived superoxide by reverse electron transport at complex I that regulates IL-1β release during NLRP3 inflammasome activation. |
|---|---|
| Συγγραφείς: | Casey AM; MRC Mitochondrial Biology Unit, Biomedical Campus, University of Cambridge, Cambridge, UK., Ryan DG; MRC Mitochondrial Biology Unit, Biomedical Campus, University of Cambridge, Cambridge, UK., Prag HA; Department of Medicine, University of Cambridge, Cambridge, UK., Chowdhury SR; MRC Mitochondrial Biology Unit, Biomedical Campus, University of Cambridge, Cambridge, UK., Marques E; MRC Mitochondrial Biology Unit, Biomedical Campus, University of Cambridge, Cambridge, UK., Turner K; MRC Mitochondrial Biology Unit, Biomedical Campus, University of Cambridge, Cambridge, UK., Gruszczyk AV; MRC Mitochondrial Biology Unit, Biomedical Campus, University of Cambridge, Cambridge, UK.; Department of Surgery and Cambridge NIHR Biomedical Research Centre, Biomedical Campus, University of Cambridge, Cambridge, UK., Yang M; University of Cologne, Faculty of Medicine and University Hospital Cologne, Institute for Metabolomics in Ageing, Cluster of Excellence Cellular Stress Responses in Aging-associated Diseases (CECAD), Cologne, Germany.; University of Cologne, Faculty of Mathematics and Natural Sciences, Institute of Genetics, Cluster of Excellence Cellular Stress Responses in Aging-associated Diseases (CECAD), Cologne, Germany., Wolf DM; MRC Mitochondrial Biology Unit, Biomedical Campus, University of Cambridge, Cambridge, UK., Miljkovic JL; MRC Mitochondrial Biology Unit, Biomedical Campus, University of Cambridge, Cambridge, UK., Valadares J; MRC Mitochondrial Biology Unit, Biomedical Campus, University of Cambridge, Cambridge, UK., Chinnery PF; MRC Mitochondrial Biology Unit, Biomedical Campus, University of Cambridge, Cambridge, UK., Hartley RC; WestCHEM School of Chemistry, University of Glasgow, Glasgow, UK., Frezza C; University of Cologne, Faculty of Medicine and University Hospital Cologne, Institute for Metabolomics in Ageing, Cluster of Excellence Cellular Stress Responses in Aging-associated Diseases (CECAD), Cologne, Germany.; University of Cologne, Faculty of Mathematics and Natural Sciences, Institute of Genetics, Cluster of Excellence Cellular Stress Responses in Aging-associated Diseases (CECAD), Cologne, Germany., Prudent J; MRC Mitochondrial Biology Unit, Biomedical Campus, University of Cambridge, Cambridge, UK., Murphy MP; MRC Mitochondrial Biology Unit, Biomedical Campus, University of Cambridge, Cambridge, UK. mpm@mrc-mbu.cam.ac.uk.; Department of Medicine, University of Cambridge, Cambridge, UK. mpm@mrc-mbu.cam.ac.uk. |
| Πηγή: | Nature metabolism [Nat Metab] 2025 Mar; Vol. 7 (3), pp. 493-507. Date of Electronic Publication: 2025 Feb 19. |
| Τύπος έκδοσης: | Journal Article |
| Γλώσσα: | English |
| Στοιχεία περιοδικού: | Publisher: Springer Nature Country of Publication: Germany NLM ID: 101736592 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2522-5812 (Electronic) Linking ISSN: 25225812 NLM ISO Abbreviation: Nat Metab Subsets: MEDLINE |
| Imprint Name(s): | Original Publication: Berlin : Springer Nature, [2019]- |
| Ιατρικοί όροι (MeSH): | NLR Family, Pyrin Domain-Containing 3 Protein*/metabolism , Macrophages*/metabolism , Interleukin-1beta*/metabolism , Inflammasomes*/metabolism , Superoxides*/metabolism , Mitochondria*/metabolism , Electron Transport Complex I*/metabolism, Reactive Oxygen Species/metabolism ; Lipopolysaccharides/pharmacology ; Animals ; Mice ; Electron Transport ; Mice, Inbred C57BL |
| Περίληψη: | Macrophages stimulated by lipopolysaccharide (LPS) generate mitochondria-derived reactive oxygen species (mtROS) that act as antimicrobial agents and redox signals; however, the mechanism of LPS-induced mitochondrial superoxide generation is unknown. Here we show that LPS-stimulated bone-marrow-derived macrophages produce superoxide by reverse electron transport (RET) at complex I of the electron transport chain. Using chemical biology and genetic approaches, we demonstrate that superoxide production is driven by LPS-induced metabolic reprogramming, which increases the proton motive force (∆p), primarily as elevated mitochondrial membrane potential (Δψ (© 2025. The Author(s).) |
| Competing Interests: | Competing interests: The authors declare no competing interests. |
| References: | Mosser, D. M. & Edwards, J. P. Exploring the full spectrum of macrophage activation. Nat. Rev. Immunol. 8, 958–969 (2008). (PMID: 19029990272499110.1038/nri2448) West, A. P. et al. TLR signalling augments macrophage bactericidal activity through mitochondrial ROS. Nature 472, 476–480 (2011). (PMID: 21525932346053810.1038/nature09973) Carneiro, F. R. G., Lepelley, A., Seeley, J. J., Hayden, M. S. & Ghosh, S. An essential role for ECSIT in mitochondrial complex I assembly and mitophagy in macrophages. Cell Rep. 22, 2654–2666 (2018). (PMID: 29514094590998910.1016/j.celrep.2018.02.051) Mills, E. L. et al. Succinate dehydrogenase supports metabolic repurposing of mitochondria to drive inflammatory macrophages. Cell 167, 457–470.e13 (2016). (PMID: 27667687586395110.1016/j.cell.2016.08.064) Kravchuk, V. et al. A universal coupling mechanism of respiratory complex I. Nature https://doi.org/10.1038/s41586-022-05199-7 (2022). Murphy, M. P. How mitochondria produce reactive oxygen species. Biochem. J. 417, 1–13 (2009). (PMID: 1906148310.1042/BJ20081386) Chouchani, E. T. et al. A unifying mechanism for mitochondrial superoxide production during ischemia-reperfusion injury. Cell Metab. 23, 254–263 (2016). (PMID: 2677768910.1016/j.cmet.2015.12.009) Tannahill, G. M. et al. Succinate is an inflammatory signal that induces IL-1β through HIF-1α. Nature 496, 238–242 (2013). (PMID: 23535595403168610.1038/nature11986) Willenborg, S. et al. Mitochondrial metabolism coordinates stage-specific repair processes in macrophages during wound healing. Cell Metab. 33, 2398–2414.e9 (2021). (PMID: 3471503910.1016/j.cmet.2021.10.004) Seim, G. L. et al. Two-stage metabolic remodelling in macrophages in response to lipopolysaccharide and interferon-γ stimulation. Nat Metab. 1, 731–742 (2019). (PMID: 32259027710880310.1038/s42255-019-0083-2) Shchepinova, M. M. et al. MitoNeoD: a mitochondria-targeted superoxide probe. Cell Chem. Biol. 24, 1285–1298.e12 (2017). (PMID: 28890317627887010.1016/j.chembiol.2017.08.003) Robb, E. L. et al. Selective superoxide generation within mitochondria by the targeted redox cycler MitoParaquat. Free Radical Biol. Med. 89, 883–894 (2015). (PMID: 10.1016/j.freeradbiomed.2015.08.021) Tur, J. et al. Mitofusin 2 in macrophages links mitochondrial ROS production, cytokine release, phagocytosis, autophagy, and bactericidal activity. Cell Rep. 32, 108079 (2020). (PMID: 3284613610.1016/j.celrep.2020.108079) Yu, W. et al. Stat2-Drp1 mediated mitochondrial mass increase is necessary for pro-inflammatory differentiation of macrophages. Redox Biol. 37, 101761 (2020). (PMID: 33080440757580310.1016/j.redox.2020.101761) Kapetanovic, R. et al. Lipopolysaccharide promotes Drp1‐dependent mitochondrial fission and associated inflammatory responses in macrophages. Immunol. Cell Biol. 98, 528–539 (2020). (PMID: 32686869749722410.1111/imcb.12363) Van den Bossche, J., O’Neill, L. A. & Menon, D. Macrophage immunometabolism: where are we (going)? Trends Immunol. 38, 395–406 (2017). (PMID: 2839607810.1016/j.it.2017.03.001) Palsson-McDermott, E. M. et al. Pyruvate kinase M2 regulates Hif-1α activity and IL-1β induction, and is a critical determinant of the Warburg effect in LPS-activated macrophages. Cell Metab. 21, 65–80 (2015). (PMID: 25565206519883510.1016/j.cmet.2014.12.005) Degrandi, D., Hoffmann, R., Beuter-Gunia, C. & Pfeffer, K. The proinflammatory cytokine-induced IRG1 protein associates with mitochondria. J. Interferon Cytokine Res. 29, 55–68 (2009). (PMID: 1901433510.1089/jir.2008.0013) Palmieri, E. M. et al. Nitric oxide orchestrates metabolic rewiring in M1 macrophages by targeting aconitase 2 and pyruvate dehydrogenase. Nat. Commun. 11, 698 (2020). (PMID: 32019928700072810.1038/s41467-020-14433-7) Perry, S. W., Norman, J. P., Barbieri, J., Brown, E. B., & Gelbard, H. A. Mitochondrial membrane potential probes and the proton gradient: a practical usage guide. BioTechniques https://doi.org/10.2144/000113610 (2011). Goehring, I. et al. Plasma membrane potential oscillations in insulin secreting Ins-1 832/13 cells do not require glycolysis and are not initiated by fluctuations in mitochondrial bioenergetics. J. Biol. Chem. 287, 15706–15717 (2012). (PMID: 22418435334607310.1074/jbc.M111.314567) Burger, N. et al. A sensitive mass spectrometric assay for mitochondrial CoQ pool redox state in vivo. Free Radic. Biol. Med. 147, 37–47 (2020). (PMID: 31811922697516710.1016/j.freeradbiomed.2019.11.028) Szibor, M. et al. Broad AOX expression in a genetically tractable mouse model does not disturb normal physiology. Dis. Model Mech. 10, 163–171 (2017). (PMID: 280676265312010) Miyadera, H. et al. Atpenins, potent and specific inhibitors of mitochondrial complex II (succinate-ubiquinone oxidoreductase). Proc. Natl Acad. Sci. USA 100, 473–477 (2003). (PMID: 1251585914101910.1073/pnas.0237315100) Sun, F. et al. Crystal structure of mitochondrial respiratory membrane protein complex II. Cell 121, 1043–1057 (2005). (PMID: 1598995410.1016/j.cell.2005.05.025) Prag, H. A. et al. Ester prodrugs of malonate with enhanced intracellular delivery protect against cardiac ischemia-reperfusion injury in vivo. Cardiovasc. Drugs Ther. 36, 1–13 (2022). (PMID: 3264816810.1007/s10557-020-07033-6) Langston, P. K. et al. Glycerol phosphate shuttle enzyme GPD2 regulates macrophage inflammatory responses. Nat. Immunol. 20, 1186–1195 (2019). (PMID: 31384058670785110.1038/s41590-019-0453-7) Cadenas, E., Boveris, A., Ragan, C. I. & Stoppani, A. O. M. Production of superoxide radicals and hydrogen peroxide by NADH-ubiquinone reductase and ubiquinol-cytochrome c reductase from beef-heart mitochondria. Arch. Biochem. Biophys. 180, 248–257 (1977). (PMID: 19552010.1016/0003-9861(77)90035-2) McManus, M. J. et al. Mitochondrial DNA variation dictates expressivity and progression of nuclear DNA mutations causing cardiomyopathy. Cell Metab. 29, 78–90.e5 (2019). (PMID: 3017430910.1016/j.cmet.2018.08.002) Lin, C. S. et al. Mouse mtDNA mutant model of Leber hereditary optic neuropathy. Proc. Natl Acad. Sci. USA 109, 20065–20070 (2012). (PMID: 23129651352387310.1073/pnas.1217113109) Malfatti, E. et al. Novel mutations of ND genes in complex I deficiency associated with mitochondrial encephalopathy. Brain 130, 1894–1904 (2007). (PMID: 1753583210.1093/brain/awm114) Yin, Z. et al. Structural basis for a complex I mutation that blocks pathological ROS production. Nat. Commun. 12, 707 (2021). (PMID: 33514727784674610.1038/s41467-021-20942-w) Arango Duque, G. & Descoteaux, A. Macrophage cytokines: involvement in immunity and infectious diseases. Front. Immunol. 5, 491 (2014). (PMID: 25339958418812510.3389/fimmu.2014.00491) de Waal Malefyt, R., Abrams, J., Bennett, B., Figdor, C. G. & de Vries, J. E. Interleukin 10(IL-10) inhibits cytokine synthesis by human monocytes: an autoregulatory role of IL-10 produced by monocytes. J. Exp. Med. 174, 1209–1220 (1991). (PMID: 194079910.1084/jem.174.5.1209) Du, C. & Sriram, S. Mechanism of inhibition of LPS-induced IL-12p40 production by IL-10 and TGF-β in ANA-1 cells. J. Leukocyte Biol. 64, 92–97 (1998). (PMID: 966528110.1002/jlb.64.1.92) Snijders, A. et al. Regulation of bioactive IL-12 production in lipopolysaccharide-stimulated human monocytes is determined by the expression of the p35 subunit. J. Immunol. 156, 1207–1212 (1996). (PMID: 855799910.4049/jimmunol.156.3.1207) Zhou, R., Yazdi, A. S., Menu, P. & Tschopp, J. A role for mitochondria in NLRP3 inflammasome activation. Nature 469, 221–225 (2011). (PMID: 2112431510.1038/nature09663) Tschopp, J. & Schroder, K. NLRP3 inflammasome activation: the convergence of multiple signalling pathways on ROS production? Nat. Rev. Immunol. 10, 210–215 (2010). (PMID: 2016831810.1038/nri2725) Juliana, C. et al. Non-transcriptional priming and deubiquitination regulate NLRP3 inflammasome activation. J. Biol. Chem. 287, 36617–36622 (2012). (PMID: 22948162347632710.1074/jbc.M112.407130) Shimada, K. et al. Oxidized mitochondrial DNA activates the NLRP3 inflammasome during apoptosis. Immunity 36, 401–414 (2012). (PMID: 22342844331298610.1016/j.immuni.2012.01.009) Groß, C. J. et al. K<sup>+</sup> efflux-independent NLRP3 inflammasome activation by small molecules targeting mitochondria. Immunity 45, 761–773 (2016). (PMID: 2769261210.1016/j.immuni.2016.08.010) Zhong, Z. et al. New mitochondrial DNA synthesis enables NLRP3 inflammasome activation. Nature 560, 198–203 (2018). (PMID: 30046112632930610.1038/s41586-018-0372-z) Du, G. et al. ROS-dependent S-palmitoylation activates cleaved and intact gasdermin D. Nature https://doi.org/10.1038/s41586-024-07373-5 (2024). Swanson, K. V., Deng, M. & Ting, J. P.-Y. The NLRP3 inflammasome: molecular activation and regulation to therapeutics. Nat. Rev. Immunol. 19, 477–489 (2019). (PMID: 31036962780724210.1038/s41577-019-0165-0) Lamkanfi, M. & Dixit, V. M. Mechanisms and functions of inflammasomes. Cell 157, 1013–1022 (2014). (PMID: 2485594110.1016/j.cell.2014.04.007) Murphy, M. P. Newly made mitochondrial DNA drives inflammation. Nature 560, 176–177 (2018). (PMID: 3007637610.1038/d41586-018-05764-z) Billingham, L. K. et al. Mitochondrial electron transport chain is necessary for NLRP3 inflammasome activation. Nat. Immunol. 23, 692–704 (2022). (PMID: 35484407909838810.1038/s41590-022-01185-3) Devant, P. et al. Gasdermin D pore-forming activity is redox-sensitive. Cell Rep. 42, 112008 (2023). (PMID: 36662620994791910.1016/j.celrep.2023.112008) Miao, R. et al. Gasdermin D permeabilization of mitochondrial inner and outer membranes accelerates and enhances pyroptosis. Immunity 56, 2523–2541.e8 (2023). (PMID: 379248121087257910.1016/j.immuni.2023.10.004) Martinon, F., Pétrilli, V., Mayor, A., Tardivel, A. & Tschopp, J. Gout-associated uric acid crystals activate the NALP3 inflammasome. Nature 440, 237–241 (2006). (PMID: 1640788910.1038/nature04516) Elliott, E. I. et al. Cutting edge: mitochondrial assembly of the NLRP3 inflammasome complex is initiated at priming. J. Immunol. 200, 3047–3052 (2018). (PMID: 2960277210.4049/jimmunol.1701723) Kim, J., Minkler, P. E., Salomon, R. G., Anderson, V. E. & Hoppel, C. L. Cardiolipin: characterization of distinct oxidized molecular species. J. Lipid Res. 52, 125–135 (2011). (PMID: 20858593299992510.1194/jlr.M010520) Baik, S. H. et al. Hexokinase dissociation from mitochondria promotes oligomerization of VDAC that facilitates NLRP3 inflammasome assembly and activation. Sci. Immunol. 8, eade7652 (2023). (PMID: 373273211036040810.1126/sciimmunol.ade7652) Bambouskova, M. et al. Itaconate confers tolerance to late NLRP3 inflammasome activation. Cell Rep. 34, 108756 (2021). (PMID: 33691097803986410.1016/j.celrep.2021.108756) Hooftman, A. et al. Macrophage fumarate hydratase restrains mtRNA-mediated interferon production. Nature 615, 490–498 (2023). (PMID: 368902271041130010.1038/s41586-023-05720-6) Liu, D. & Huang, Y. Protocol for in vivo and in vitro activation of NLRP3 inflammasome in mice using monosodium urate. STAR Protoc. 4, 102554 (2023). (PMID: 376827171049358510.1016/j.xpro.2023.102554) Iyer, S. S. et al. Mitochondrial cardiolipin is required for Nlrp3 inflammasome activation. Immunity 39, 311–323 (2013). (PMID: 23954133377928510.1016/j.immuni.2013.08.001) Dostert, C. et al. Innate immune activation through Nalp3 Inflammasome sensing of asbestos and silica. Science 320, 674–677 (2008). (PMID: 18403674239658810.1126/science.1156995) Prag, H. A. et al. Ischemia-selective cardioprotection by malonate for ischemia/reperfusion injury. Circ. Res. 131, 528–541 (2022). (PMID: 35959683942674210.1161/CIRCRESAHA.121.320717) Pang, Z. et al. MetaboAnalyst 5.0: narrowing the gap between raw spectra and functional insights. Nucleic Acids Res. 49, W388–W396 (2021). (PMID: 34019663826518110.1093/nar/gkab382) Strehler, B. L. in Methods of Enzymatic Analysis 2nd edn (ed. Bergmeyer, H. U.) 2112–2126 (Academic Press, 1974). Gruszczyk, A. V. et al. Mitochondrial metabolism and bioenergetic function in an anoxic isolated adult mouse cardiomyocyte model of in vivo cardiac ischemia-reperfusion injury. Redox Biol. 54, 102368 (2022). (PMID: 35749842923447210.1016/j.redox.2022.102368) Burr, S. P. & Chinnery, P. F. Measuring single-cell mitochondrial DNA copy number and heteroplasmy using digital droplet polymerase chain reaction. J. Vis. Exp. https://doi.org/10.3791/63870 (2022). Tilokani, L. et al. AMPK-dependent phosphorylation of MTFR1L regulates mitochondrial morphology. Sci. Adv. 8, eabo7956 (2022). (PMID: 36367943965186510.1126/sciadv.abo7956) Zhang, Y. et al. Neuronal TORC1 modulates longevity via AMPK and cell nonautonomous regulation of mitochondrial dynamics in C. elegans. eLife 8, e49158 (2019). (PMID: 31411562671350910.7554/eLife.49158) |
| Grant Information: | MC_UU_00028/4 RCUK | Medical Research Council (MRC); MC_UU_00014/5 RCUK | Medical Research Council (MRC); MC_UU_00028/5 RCUK | Medical Research Council (MRC); MC_UU_00015/3 RCUK | Medical Research Council (MRC); 220257/Z/20/Z Wellcome Trust (Wellcome); United Kingdom WT_ Wellcome Trust; ALTF 828-2021 European Molecular Biology Organization (EMBO) |
| Substance Nomenclature: | 0 (NLR Family, Pyrin Domain-Containing 3 Protein) 0 (Interleukin-1beta) 0 (Inflammasomes) 11062-77-4 (Superoxides) EC 7.1.1.2 (Electron Transport Complex I) 0 (Nlrp3 protein, mouse) 0 (Reactive Oxygen Species) 0 (Lipopolysaccharides) |
| Entry Date(s): | Date Created: 20250219 Date Completed: 20250509 Latest Revision: 20250509 |
| Update Code: | 20260130 |
| PubMed Central ID: | PMC11946910 |
| DOI: | 10.1038/s42255-025-01224-x |
| PMID: | 39972217 |
| Βάση Δεδομένων: | MEDLINE |
| ISSN: | 2522-5812 |
|---|---|
| DOI: | 10.1038/s42255-025-01224-x |