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Oxidized arachidonic and adrenic PEs navigate cells to ferroptosis

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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Figure 1: Oxygenation of esterified AA contributes to RSL3-induced ferroptosis in WT and Acsl4 KO Pfa1 cells.
Figure 2: Effects of exogenous AA on RSL3-triggered ferroptosis.
Figure 3: Screening of phospholipids and their oxidation products identifies ferroptosis death signals.
Figure 4: Oxygenated PE species identified in ferroptotic Gpx4 KO cells and kidney of Gpx4 KO mice.
Figure 5: Labeling with AA-d8 unravels pathways leading to oxygenated diacylated PE ferroptotic signals.
Figure 6: A 15-LOX phospholipid oxidation product, 15-hydroperoxy-SAPE, triggers ferroptosis in WT and Acsl4 KO Pfa1 cells.

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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.

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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.

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

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Kagan, V., Mao, G., Qu, F. et al. Oxidized arachidonic and adrenic PEs navigate cells to ferroptosis. Nat Chem Biol 13, 81–90 (2017). https://doi.org/10.1038/nchembio.2238

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