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Global survey of cell death mechanisms reveals metabolic regulation of ferroptosis

Nature Chemical Biology volume 12, pages 497–503 (2016)Cite this article

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Abstract

Apoptosis is one type of programmed cell death. Increasingly, non-apoptotic cell death is recognized as being genetically controlled, or 'regulated'. However, the full extent and diversity of alternative cell death mechanisms remain uncharted. Here we surveyed the landscape of pharmacologically accessible cell death mechanisms. In an examination of 56 caspase-independent lethal compounds, modulatory profiling showed that 10 compounds induced three different types of regulated non-apoptotic cell death. Optimization of one of those ten resulted in the discovery of FIN56, a specific inducer of ferroptosis. Ferroptosis has been found to occur when the lipid-repair enzyme GPX4 is inhibited. FIN56 promoted degradation of GPX4. FIN56 also bound to and activated squalene synthase, an enzyme involved in isoprenoid biosynthesis, independent of GPX4 degradation. These discoveries show that dysregulation of lipid metabolism is associated with ferroptosis. This systematic approach is a means to discover and characterize novel cell death phenotypes.

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Figure 1: Modulatory profiling revealed three types of regulated non-apoptotic cell death.
Figure 2: Optimization of CIL56 revealed a potent and selective ferroptosis inducer.
Figure 3: FIN56-induced ferroptosis decreases GPX4 expression.
Figure 4: SQS encoded by FDFT1 as FIN56's target protein.
Figure 5: Validating SQS as the functionally relevant target for FIN56's lethality.
Figure 6: ACC inhibitor prevents GPX4 protein degradation.

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Acknowledgements

We thank H. Li (Columbia University, New York, New York, USA) and C. Henderson (Biogen Idec, Cambridge, Massachusetts, USA) for providing mevalonic acid and farnesyl pyrophosphate, C.-I. Liu and A.H.J. Wang (Academia Sinica, Taipei, Taiwan) for the truncated human squalene synthase construct, R. Weinberg (Whitehead Institute, Cambridge, Massachusetts, USA) for engineered BJ cell lines (BJeLR, DRD, BJeHLT, and BJeH), E. Schon (Columbia University, New York, New York, USA) for 143B cells, E. Lee for assistance with ACC experiments, and V. Viswanathan for helpful discussions. This research was funded by the Howard Hughes Medical Institute, the US National Institutes of Health (grants 5R01CA097061, 5R01GM085081, and R01CA161061 to B.R.S.), New York Stem Cell Science (grant C026715 to B.R.S.) and the US National Cancer Institute (K99 Pathway to Independence Award 1K99CA166517-01 to S.J.D.).

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Author notes

  1. Rachid Skouta, Wan Seok Yang, Scott J Dixon, Carlos A Valenzuela & Adam J Wolpaw

    Present address: Present addresses: Department of Chemistry, Border Biomedical Research Center, the University of Texas at El Paso, El Paso, Texas, USA (R.S. and C.A.V.); Department of Biological Sciences, St. John's University, Queens, New York, USA (W.S.Y.); Department of Biology, Stanford University, Stanford, California, USA (S.J.D.); Divisions of Hematology and Oncology, the Children's Hospital of Philadelphia, Philadelphia, Pennsylvania, USA (A.J.W.).,

Authors and Affiliations

  1. Department of Biological Sciences, Columbia University, New York, New York, USA

    Kenichi Shimada, Rachid Skouta, Anna Kaplan, Wan Seok Yang, Scott J Dixon, Adam J Wolpaw & Brent R Stockwell

  2. Department of Pharmacology, Columbia University, New York, New York, USA

    Miki Hayano

  3. Quantitative Proteomics Center, Columbia University, New York, New York, USA

    Lewis M Brown

  4. Department of Chemistry, Columbia University, New York, New York, USA

    Brent R Stockwell

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Contributions

K.S. and B.R.S. conceived of the project, designed the experiments, analyzed the data, and wrote the manuscript. K.S. performed all experiments and analyses except the following: R.S. and C.A.V. synthesized CIL56 analogs; A.K. expressed and purified SQS protein and performed confirmatory SQS pulldown experiments and competition assays; W.S.Y. performed the GPX4 enzymatic assay; and M.H. performed confirmatory siRNA experiments. L.M.B. performed proteomic analysis for target identification. S.J.D. participated in characterization of TOFA's effect on FIN56. A.J.W. assisted in modulatory profiling experiments. S.J.D. assisted in writing the manuscript.

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Correspondence to Brent R Stockwell.

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Supplementary information

Supplementary Text and Figures

Supplementary Results, Supplementary Figures 1–14 and Supplementary Tables 1–3. (PDF 5029 kb)

Supplementary Data Set 1

Lists of lethal compounds, chemical death modulators, cell lines, and modulatory profiles of characterized and uncharacterized lethal compounds. (XLSX 104 kb)

Supplementary Note 1

Supplementary Note 1 (PDF 2402 kb)

Supplementary Note 2

Synthetic Procedures (PDF 1066 kb)

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Shimada, K., Skouta, R., Kaplan, A. et al. Global survey of cell death mechanisms reveals metabolic regulation of ferroptosis. Nat Chem Biol 12, 497–503 (2016). https://doi.org/10.1038/nchembio.2079

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