Two-stage metabolic remodelling in macrophages in response to lipopolysaccharide and interferon-γ stimulation

Abstract

In response to signals associated with infection or tissue damage, macrophages undergo a series of dynamic phenotypic changes. Here we show that during the response to lipopolysaccharide and interferon-γ stimulation, metabolic reprogramming in macrophages is also highly dynamic. Specifically, the tricarboxylic acid cycle undergoes a two-stage remodelling: the early stage is characterized by a transient accumulation of intermediates including succinate and itaconate, whereas the late stage is marked by the subsidence of these metabolites. The metabolic transition into the late stage is largely driven by the inhibition of the pyruvate dehydrogenase complex (PDHC) and the oxoglutarate dehydrogenase complex (OGDC), which is controlled by the dynamic changes in the lipoylation state of both PDHC and OGDC E2 subunits and phosphorylation of the PDHC E1 subunit. This dynamic metabolic reprogramming results in a transient metabolic state that strongly favours hypoxia-inducible factor-1α (HIF-1α) stabilization during the early stage, which subsides by the late stage; consistently, HIF-1α levels follow this trend. This study elucidates a dynamic and mechanistic picture of metabolic reprogramming in lipopolysaccharide and interferon-γ stimulated macrophages, and provides insights into how changing metabolism can regulate the functional transitions in macrophages over the course of an immune response.

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Fig. 1: Macrophages undergo dynamic metabolomic and functional changes in response to LPS and IFN-γ stimulation.
Fig. 2: Glucose labelling reveals dynamic reprogramming of TCA cycle flux in LPS and IFN-γ stimulated macrophages.
Fig. 3: Glutamine labelling reveals remodelling of TCA cycle flux at the late stage.
Fig. 4: Activities of PDHC and OGDC decrease in response to LPS and IFN-γ exposure.
Fig. 5: Changes in key metabolite levels correlate with changes in HIF-1α protein and histone methylation.
Fig. 6: Dynamic regulation of PDHC and OGDC in LPS and IFN-γ stimulated macrophages.
Fig. 7: Alternative substrate use in LPS and IFN-γ stimulated macrophages.
Fig. 8: Two-stage remodelling of the TCA cycle in LPS and IFN-γ stimulated macrophages.

Data availability

The data that support the findings of this study are available from the corresponding author upon request.

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Acknowledgements

This work is primarily supported by the Morgridge Institute for Research (start-up fund for J.F.). Additionally, R.S.E. is supported by NIH grant no. R01DK66600, D.J.P. is supported by NIH grant no. R35GM130294. The authors would like to thank the University of Wisconsin Carbone Cancer Center. This is supported in part by NIH/NCI grant no. P30CA014520—UW Cancer Center Support Grant. The authors would also like to thank Yatrik Shah at the University of Michigan for sharing the ODD-luc mice.

Author information

J.F. and G.L.S. designed the study and analyzed data. E.C.B. performed and analyzed data from DCA treatment experiments. S.V.J. carried out and analyzed data from palmitic-acid labeling studies. F.J.Y. performed some cytokine assays and qPCR. A.R.J. carried out the isolation and analysis of histones. G.L.S. performed all remaining experiments. R.S.E. contributed to the investigation of the role of metabolites in regulating HIF-1α. D.J.P. contributed to the identification of changing lipoylation as a mechanism for PDHC and OGDC regulation. G.L.S. and J.F. wrote the manuscript. R.S.E. and D.J.P. edited the manuscript.

Correspondence to Jing Fan.

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