Abstract

Enigmatic lipid peroxidation products have been claimed as the proximate executioners of ferroptosis—a specialized death program triggered by insufficiency of glutathione peroxidase 4 (GPX4). Using quantitative redox lipidomics, reverse genetics, bioinformatics and systems biology, we discovered that ferroptosis involves a highly organized oxygenation center, wherein oxidation in endoplasmic-reticulum-associated compartments occurs on only one class of phospholipids (phosphatidylethanolamines (PEs)) and is specific toward two fatty acyls—arachidonoyl (AA) and adrenoyl (AdA). Suppression of AA or AdA esterification into PE by genetic or pharmacological inhibition of acyl-CoA synthase 4 (ACSL4) acts as a specific antiferroptotic rescue pathway. Lipoxygenase (LOX) generates doubly and triply-oxygenated (15-hydroperoxy)-diacylated PE species, which act as death signals, and tocopherols and tocotrienols (vitamin E) suppress LOX and protect against ferroptosis, suggesting a homeostatic physiological role for vitamin E. This oxidative PE death pathway may also represent a target for drug discovery.

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Acknowledgements

We thank J. Ruzicka (Thermo Fisher Scientific) for help in obtaining MS3 spectra of PE oxidation products using tribrid Fusion Lumos. Supported by the US National Institutes of Health (P01HL114453 to R.K.M., U19AI068021 to J.G., NS076511 to V.E.K., NS061817 to H.B., P41GM103712 to I.B. and ES020693 to Y.Y.T.), the Human Frontier Science Program (HFSP-RGP0013/2014), and the Deutsche Forschungsgemeinschaft (CO 291/2-3 and CO 291/5-1) to M.C.

Author information

Author notes

    • Gaowei Mao
    • , Feng Qu
    •  & Jose Pedro Friedmann Angeli

    These authors contributed equally to this work.

Affiliations

  1. Department of Environmental and Occupational Health, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

    • Valerian E Kagan
    • , Gaowei Mao
    • , Feng Qu
    • , Haider Hussain Dar
    • , Vladimir A Tyurin
    • , Vladimir B Ritov
    • , Alexandr A Kapralov
    • , Andrew A Amoscato
    • , Jianfei Jiang
    • , Dariush Mohammadyani
    • , Yulia Y Tyurina
    •  & Hülya Bayır
  2. Department of Pharmacology and Chemical Biology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

    • Valerian E Kagan
  3. Department of Chemistry, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

    • Valerian E Kagan
  4. Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

    • Valerian E Kagan
    •  & Joel Greenberger
  5. Department of Critical Care Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

    • Gaowei Mao
    • , Tamil Anthonymuthu
    • , Qin Yang
    •  & Hülya Bayır
  6. Institute of Developmental Genetics, Helmholtz Zentrum München, Neuherberg, Germany.

    • Jose Pedro Friedmann Angeli
    • , Sebastian Doll
    • , Bettina Proneth
    •  & Marcus Conrad
  7. Department of Cell Biology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

    • Claudette St Croix
    •  & Simon Watkins
  8. Department of Computational and Systems Biology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

    • Bing Liu
    •  & Ivet Bahar
  9. Division of Metabolic and Vascular Health, University of Warwick, Coventry, UK.

    • Judith Klein-Seetharaman
  10. Department of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

    • Rama K Mallampalli
  11. Department of Biological Sciences, Columbia University, New York, New York, USA.

    • Brent R Stockwell
  12. Department of Chemistry, Columbia University, New York, New York, USA.

    • Brent R Stockwell

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Contributions

V.E.K., M.C. and H.B. formulated the idea, designed the study and wrote the manuscript. G.M. and J.P.F.A. performed cell experiments. Y.Y.T. and F.Q. performed MS lipid analysis, interpreted data. C.S. and S.W. performed cell imaging experiments. T.A., V.A.T. and A.A.A. performed model systems experiments. D.M. and J.K.-S. performed computational modeling. B.L. and I.B. performed network analysis. S.D., H.H.D., J.J., V.B.R., A.A.K., B.P. and Q.Y. participated in cell or animal experiments. J.G., R.K.M. and B.R.S. participated in formulating the idea and writing the manuscript. All authors discussed the results and commented on the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Valerian E Kagan or Marcus Conrad or Hülya Bayır.

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https://doi.org/10.1038/nchembio.2238

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