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Abstract

Ferroptosis is a non-apoptotic form of regulated cell death caused by the failure of the glutathione-dependent lipid-peroxide-scavenging network. FINO2 is an endoperoxide-containing 1,2-dioxolane that can initiate ferroptosis selectively in engineered cancer cells. We investigated the mechanism and structural features necessary for ferroptosis initiation by FINO2. We found that FINO2 requires both an endoperoxide moiety and a nearby hydroxyl head group to initiate ferroptosis. In contrast to previously described ferroptosis inducers, FINO2 does not inhibit system xc or directly target the reducing enzyme GPX4, as do erastin and RSL3, respectively, nor does it deplete GPX4 protein, as does FIN56. Instead, FINO2 both indirectly inhibits GPX4 enzymatic function and directly oxidizes iron, ultimately causing widespread lipid peroxidation. These findings suggest that endoperoxides such as FINO2 can initiate a multipronged mechanism of ferroptosis.

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Acknowledgements

We thank C. Lin for assistance with NMR spectroscopy and mass spectrometry, C. Hu for X-ray analysis, along with the Materials Research Science and Engineering Center (MRSEC) program of the National Science Foundation (NSF) under Award Numbers DMR-0820341 and DMR-1420073, and J. Chung for assistance with cell culture. This research was supported by the Training Program in Molecular Biophysics Grant (T32GM008281 to M.M.G.), the National Cancer Institute (R35CA209896 and P01CA087497 to B.R.S), the National Institute of General Medical Sciences (1RO1GM118730 to K.A.W.), the National Heart, Lung, and Blood Institute (HL114453 to V.E.K. and Y.Y.T.), the National Institute of Allergy and Infectious Diseases (U19AI068021 to V.E.K. and Y.Y.T.) and the MRSEC Program of the National Science Foundation (DMR-1420073 to E.P.-P.). The Bruker Avance-400, 500 and 600 MHz spectrometers (NYU) were acquired through the support of the National Science Foundation (CHE-01162222).

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Author notes

  1. These authors contributed equally: Michael M. Gaschler and Alexander A. Andia.

Affiliations

  1. Department of Chemistry, Columbia University, New York, NY, USA

    • Michael M. Gaschler
    • , Hengrui Liu
    •  & Brent R. Stockwell
  2. Department of Chemistry, New York University, New York, NY, USA

    • Alexander A. Andia
    • , Brisa Hurlocker
    • , Christopher A. Vaiana
    • , Daniel W. Heindel
    • , Dylan S. Zuckerman
    • , Amy Y. Chan
    • , Eveliz Peguero-Pereira
    • , Lara K. Mahal
    •  & K. A. Woerpel
  3. Department of Biological Sciences, Columbia University, New York, NY, USA

    • Joleen M. Csuka
    • , Pieter H. Bos
    • , Eduard Reznik
    • , Ling F. Ye
    • , Annie J. Lin
    •  & Brent R. Stockwell
  4. Department of Environmental and Occupational Health, University of Pittsburgh, Pittsburgh, PA, USA

    • Yulia Y. Tyurina
    •  & Valerian E. Kagan
  5. Retrotope Inc, Los Altos, CA, USA

    • Mikhail S. Shchepinov
  6. Department of Chemistry, Belarusian State University, Minsk, Belarus

    • Andrei V. Bekish
  7. Institute of Physical Organic Chemistry, National Academy of Sciences of Belarus, Minsk, Belarus

    • Maksim A. Fomich
    •  & Vadim V. Shmanai
  8. Department of Pharmacology, Columbia University, New York, NY, USA

    • Jacob. D. Daniels

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Contributions

M.M.G., A.A.A., H.L., B.R.S. and K.A.W. contributed to the writing of the manuscript. M.M.G., A.A.A., L.K.M., V.E.K., B.R.S. and K.A.W. designed and planned research. M.M.G., A.A.A., H.L., J.M.C., D.W.H., D.S.Z., P.H.B., Y.Y.T. and J.D.D. conducted in vitro biochemical and metabolomic assays. M.M.G, A.A.A., B.H., C.A.V., D.W.H. and A.J.L. collected and analyzed cell viability data. L.F.Y. performed quantitative PCR. A.A.A. and H.L. conducted NMR studies. A.A.A. conducted stability studies. H.L. and E.R. conducted western blotting experiments. A.A.A., B.H. and D.S.Z. synthesized FINO2 and all structural analogues. A.Y.C. aided in furan synthesis. E.P.-P. aided in oxetane synthesis. M.A.F., A.V.B., V.V.S., A.J.L. and M.S.S. synthesized deuterated arachidonic acids. All authors have given their approval of the final version of the manuscript.

Competing interests

M.S.S. is the Chief Scientific Officer of Retrotrope, Inc.

Corresponding authors

Correspondence to K. A. Woerpel or Brent R. Stockwell.

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https://doi.org/10.1038/s41589-018-0031-6

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