Redox lipid reprogramming commands susceptibility of macrophages and microglia to ferroptotic death


Ferroptotic death is the penalty for losing control over three processes—iron metabolism, lipid peroxidation and thiol regulation—that are common in the pro-inflammatory environment where professional phagocytes fulfill their functions and yet survive. We hypothesized that redox reprogramming of 15-lipoxygenase (15-LOX) during the generation of pro-ferroptotic signal 15-hydroperoxy-eicosa-tetra-enoyl-phosphatidylethanolamine (15-HpETE-PE) modulates ferroptotic endurance. Here, we have discovered that inducible nitric oxide synthase (iNOS)/NO-enrichment of activated M1 (but not alternatively activated M2) macrophages/microglia modulates susceptibility to ferroptosis. Genetic or pharmacologic depletion/inactivation of iNOS confers sensitivity on M1 cells, whereas NO donors empower resistance of M2 cells to ferroptosis. In vivo, M1 phagocytes, in comparison to M2 phagocytes, exert higher resistance to pharmacologically induced ferroptosis. This resistance is diminished in iNOS-deficient cells in the pro-inflammatory conditions of brain trauma or the tumour microenvironment. The nitroxygenation of eicosatetraenoyl (ETE)-PE intermediates and oxidatively truncated species by NO donors and/or suppression of NO production by iNOS inhibitors represent a novel redox mechanism of regulation of ferroptosis in pro-inflammatory conditions.

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Fig. 1: Differential sensitivity of activated M1 and alternatively activated M2 macrophages and microglia to RSL3-induced ferroptosis.
Fig. 2: Sensitivity of activated (M1) macrophages and microglial cells to RSL3-induced ferroptosis depends on the levels of iNOS expression.
Fig. 3: NO protects cells against RSL3-induced ferroptosis.
Fig. 4: NO suppresses RSL3-induced accumulation of oxidatively modified PE species in alternatively activated (M2) RAW 264.7 macrophages.
Fig. 5: iNOS/NO-driven mechanisms of ferroptosis regulation in vivo.
Fig. 6: NO/O2 interactions with 15-LOX-2 and entry/exit pathways observed in MD simulations of 15-LOX-2 and the 15-LOX-2–PEBP1 complex.

Data availability

The raw data are available at the following link

Code used for the analysis of the MD simulations of NO interactions with 15-LOX has been made available in two formats: (1) Jupyter Notebook and (2) html (


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This work was supported by NIH (HL114453-06, U19AI068021, CA165065-06, NS076511, NS061817, P41GM103712) and by Russian academic excellence project ‘5-100’.

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V.E.K. and H.B. conceived the study. A.A.K., Q.Y., H.H.D., G.V.S., H.-C.T., M.A.A. and L.A.P. performed experiments with cells. V.E.K., H.B. and D.I.G. designed in vivo experiments. Q.Y., Y.L.W., R.K. and Y.G. performed in vivo experiments. A.A.K., Q.Y. and H.H.D. analyzed the data. Y.Y.T., T.S.A. and V.A.T. performed MS measurements and analyzed data. Y.Y.T., T.S.A. and R.M.D. discussed and interpreted MS results. K.M.-R., B.L. and I.H.S. performed computational modelling. I.B. supervised computational studies. C.M.S.C. performed imaging experiments and participated in interpreting them. H.B., D.A.S., R.K.M. and D.I.G. participated in formulating the idea and interpreting the data. P.S.T. and J.S.G. participated in the discussion and helped in writing the manuscript. Y.Y.T., I.B. and H.B. participated in writing the manuscript. H.B. and V.E.K. wrote the manuscript.

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Correspondence to Dmitry I. Gabrilovich or Hülya Bayır or Valerian E. Kagan.

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Fragment of MD simulations of competitive binding of NO and O2 molecules to 15-LOX-2.

Supplementary Information

Supplementary Figs. 1–11.

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Supplementary Tables 1 and 2

Supplementary Tables 1 and 2.

Supplementary Video 1

Fragment of MD simulations of competitive binding of NO and O2 molecules to 15-LOX-2.

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Kapralov, A.A., Yang, Q., Dar, H.H. et al. Redox lipid reprogramming commands susceptibility of macrophages and microglia to ferroptotic death. Nat Chem Biol 16, 278–290 (2020).

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