Abstract
Macrophages activated by the Gram-negative bacterial product lipopolysaccharide switch their core metabolism from oxidative phosphorylation to glycolysis1. Here we show that inhibition of glycolysis with 2-deoxyglucose suppresses lipopolysaccharide-induced interleukin-1β but not tumour-necrosis factor-α in mouse macrophages. A comprehensive metabolic map of lipopolysaccharide-activated macrophages shows upregulation of glycolytic and downregulation of mitochondrial genes, which correlates directly with the expression profiles of altered metabolites. Lipopolysaccharide strongly increases the levels of the tricarboxylic-acid cycle intermediate succinate. Glutamine-dependent anerplerosis is the principal source of succinate, although the ‘GABA (γ-aminobutyric acid) shunt’ pathway also has a role. Lipopolysaccharide-induced succinate stabilizes hypoxia-inducible factor-1α, an effect that is inhibited by 2-deoxyglucose, with interleukin-1β as an important target. Lipopolysaccharide also increases succinylation of several proteins. We therefore identify succinate as a metabolite in innate immune signalling, which enhances interleukin-1β production during inflammation.
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Change history
10 April 2013
The product code for the anti-HIF-1α antibody was corrected.
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Acknowledgements
We thank the European Research Council, Science Foundation Ireland, the Health Research Board, European Union FP7 programme ‘TIMER’, Wellcome Trust, National Institutes of Health, Helmsley Trust, Nestle Research Centre, VESKI, The Duquesne University Hunkele Dreaded Disease Award, The Interleukin Foundation and the National Health and Medical Research Council for funding. We also thank R. Thompson for assistance with Hif1a−/− mice and M. Murphy for discussions.
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G.M.T. designed and did experiments, analysed data and wrote the paper; L.A.J.O. conceived ideas and oversaw the research programme; A.M.C, E.M.P., A.F.M. and J.A. designed and did experiments and analysed data; C.F., N.J.B., B.K., N.H.F., L.Z., A.G., Z.T., S.S.J., S.C.C., S.W., K.P. and F.C.B did experiments; G.G., R.J.X., C.C., M.H. and B.E.C. performed bioinformatic analysis; E.C., V.N., M.W., C.T.T., H.L., S.L.M., E.G., V.K. and C.C., provided advice and reagents; P.E.A. and R.J.X. conceived ideas and oversaw a portion of the work.
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Tannahill, G., Curtis, A., Adamik, J. et al. Succinate is an inflammatory signal that induces IL-1β through HIF-1α. Nature 496, 238–242 (2013). https://doi.org/10.1038/nature11986
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DOI: https://doi.org/10.1038/nature11986
Christos Chinopoulos
Perhaps LPS upregulated Irg-1 expression in macrophages, yileding itaconate which is a preferred substrate for succinyl CoA ligase over succinate (when the reaction proceeds towards succinyl CoA formation), thus resulting in succinate accumulation? see Proc Natl Acad Sci U S A. 2013 May 7;110(19):7820-5 and ADLER J, WANG SF, & Lardy HA (1957) The metabolism of itaconic acid by liver mitochondria. and J. Biol. Chem, 229, 865-879 and WANG SF, ADLER J, & Lardy HA (1961) The pathway of itaconate metabolism by liver mitochondria. J. Biol. Chem, 236, 26-30.
Christos Chinopoulos
Could it be that LPS-induced succinate over-production is due to itaconate formation? LPS induces Irg1, a gene coding for cis-aconitate decarboxylase, specifically expressed in cells of macrophage lineage Proc Natl Acad Sci U S A. 2013 May 7;110(19):7820-5. doi: 10.1073/pnas.1218599110, yielding itaconate. Itaconate is preferentially used by succinyl CoA ligase forming itaconyl CoA, thus generating an accumulation of succinate when the ligase operates towards succinyl CoA formation ADLER J, WANG SF, & Lardy HA (1957) The metabolism of itaconic acid by liver mitochondria. J. Biol. Chem, 229, 865-879, and WANG SF, ADLER J, & Lardy HA (1961) The pathway of itaconate metabolism by liver mitochondria. J. Biol. Chem, 236, 26-30.