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Endogenous oxidized phospholipids reprogram cellular metabolism and boost hyperinflammation

Nature Immunology volume 21, pages 42–53 (2020)Cite this article

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Abstract

Pathogen-associated molecular patterns (PAMPs) have the capacity to couple inflammatory gene expression to changes in macrophage metabolism, both of which influence subsequent inflammatory activities. Similar to their microbial counterparts, several self-encoded damage-associated molecular patterns (DAMPs) induce inflammatory gene expression. However, whether this symmetry in host responses between PAMPs and DAMPs extends to metabolic shifts is unclear. Here, we report that the self-encoded oxidized phospholipid oxPAPC alters the metabolism of macrophages exposed to lipopolysaccharide. While cells activated by lipopolysaccharide rely exclusively on glycolysis, macrophages exposed to oxPAPC also use mitochondrial respiration, feed the Krebs cycle with glutamine, and favor the accumulation of oxaloacetate in the cytoplasm. This metabolite potentiates interleukin-1β production, resulting in hyperinflammation. Similar metabolic adaptions occur in vivo in hypercholesterolemic mice and human subjects. Drugs that interfere with oxPAPC-driven metabolic changes reduce atherosclerotic plaque formation in mice, thereby underscoring the importance of DAMP-mediated activities in pathophysiological conditions.

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Fig. 1: oxPAPC induces a hypermetabolic state in macrophages.
Fig. 2: oxPAPC promotes a hyperinflammatory phenotype in LPS-stimulated macrophages.
Fig. 3: Nitric oxide inhibition and respiration maintenance promoted by oxPAPC enables the hyperproduction of IL-1β.
Fig. 4: Glutamine is strictly required for oxPAPC-mediated hyperinflammation.
Fig. 5: oxPAPC potentiates HIF-1α through OAA accumulation.
Fig. 6: Conversion of citrate into OAA in the cytoplasm governs the induction of the hyperinflammatory phenotype in macrophages treated with oxPAPC and LPS.
Fig. 7: oxPAPC-driven immunometabolic adaptations occur in hypercholesterolemic mice.
Fig. 8: The hypermetabolism induced by oxidized phospholipids can be targeted against atherosclerosis.

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Data availability

The data that support the findings of this study are available from the corresponding author upon request. Uncropped raw immunoblot images can be found in the ‘Source data’ section of the Supplementary Information. Participant-level phenotype and genotype data from the FHS are accessible from the US National Center for Biotechnology Information database of Genotypes and Phenotypes at https://dbgap.ncbi.nlm.nih.gov/ to approved scientific investigators pursuing research questions that are consistent with the informed consent agreements provided by individual research participants. The FHS expression data are available from the database of Genotypes and Phenotypes at https://www.ncbi.nlm.nih.gov/projects/gap/cgi-bin/study.cgi?study_id=phs000363.v3.p6.

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Acknowledgements

We thank F. Granucci, J. C. Kagan and L. R. Marek for discussions, help and support. R.S. thanks the UCLA QCBio Collaboratory community, directed by M. Pellegrini. I.Z. is supported by National Institutes of Health (NIH) grants 1R01AI121066, 1R01DK115217 and NIAID-DAIT-NIHAI201700100. J.R.S. is supported by NIH grant 1R15HL121770-01A1. The FHS is funded by NIH contracts N01-HC-25195 and HHSN268201500001I. The laboratory work for the FHS investigation was funded by the Division of Intramural Research, National Heart, Lung, and Blood Institute, NIH, and by a Director’s Challenge Award, NIH (principal investigator: D.L.). This study utilized the computational resources of the Biowulf system at the NIH in Bethesda, Maryland (http://hpc.nih.gov). M.M.M. is supported by NIH grant K99HL136875. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.

Author information

Authors and Affiliations

  1. Division of Immunology and Division of Gastroenterology, Boston Children’s Hospital, Harvard Medical School, Boston, MA, USA

    Marco Di Gioia & Ivan Zanoni

  2. Institute for Quantitative and Computational Biosciences, University of California, Los Angeles, Los Angeles, CA, USA

    Roberto Spreafico

  3. Department of Chemical and Paper Engineering, Western Michigan University, Kalamazoo, MI, USA

    James R. Springstead

  4. Department of Cardiology, Boston Children’s Hospital, Boston, MA, USA

    Michael M. Mendelson

  5. Population Sciences Branch, Division of Intramural Research, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD, USA

    Roby Joehanes & Daniel Levy

  6. Department of Biotechnology and Biosciences, University of Milano–Bicocca, Milan, Italy

    Ivan Zanoni

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Contributions

M.D.G. designed, performed and analyzed the experiments. R.S. performed the statistical comparisons on the FHS aggregated data. J.R.S. produced oxPAPC and PEIPC, and participated in the analyses of data. M.M.M., R.J. and D.L. developed and analyzed the FHS expression project. I.Z. conceived the project, designed the experiments, supervised the study and wrote the paper.

Corresponding author

Correspondence to Ivan Zanoni.

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Competing interests

I.Z., M.D.G. and Boston Children’s Hospital have filed an international patent application (US patent application number 62/851,561) that relates to the metabolic activity of oxPAPC.

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Peer review information Zoltan Fehervari was the primary editor on this article and managed its editorial process and peer review in collaboration with the rest of the editorial team.

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Extended data

Extended Data Fig. 1 oxPAPC drives hyperactivation and hypermetabolism in macrophages.

Schematic depicting oxPAPC activities. oxPAPC is a mixture of oxidized phospholipids that induce an hyperinflammatory state in phagocytes upon LPS encounter and/or during atherosclerosis development. Moieties such as POVPC or PGPC contained in oxPAPC drive the formation of hyperactive cells that are characterized by inflammasome activation in the absence of pyroptosis. In contrast to POVPC or PGPC, PEIPC engages a hypermetabolic state in phagocytes that favors IL-1β accumulation and that is characterized by: i) the simultaneous activation of OXPHOS and aerobic glycolysis; ii) glutamine utilization to feed the TCA cycle; iii) oxaloacetate (OAA) accumulation in the cytoplasm to potentiate HIF-1α activation.

Supplementary information

Supplementary Information

Supplementary Figs. 1–8.

Reporting Summary

Supplementary Table 1

Metabolomic analysis of BMDMs primed for 3 h with LPS (1 μg ml−1) and treated, or not, with oxPAPC (100 μg ml−1).

Supplementary Table 2

FHS gene expression analysis.

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Di Gioia, M., Spreafico, R., Springstead, J.R. et al. Endogenous oxidized phospholipids reprogram cellular metabolism and boost hyperinflammation. Nat Immunol 21, 42–53 (2020). https://doi.org/10.1038/s41590-019-0539-2

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