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The glutathione redox system is essential to prevent ferroptosis caused by impaired lipid metabolism in clear cell renal cell carcinoma

Oncogenevolume 37pages54355450 (2018) | Download Citation

Abstract

Metabolic reprogramming is a prominent feature of clear cell renal cell carcinoma (ccRCC). Here we investigated metabolic dependencies in a panel of ccRCC cell lines using nutrient depletion, functional RNAi screening and inhibitor treatment. We found that ccRCC cells are highly sensitive to the depletion of glutamine or cystine, two amino acids required for glutathione (GSH) synthesis. Moreover, silencing of enzymes of the GSH biosynthesis pathway or glutathione peroxidases, which depend on GSH for the removal of cellular hydroperoxides, selectively reduced viability of ccRCC cells but did not affect the growth of non-malignant renal epithelial cells. Inhibition of GSH synthesis triggered ferroptosis, an iron-dependent form of cell death associated with enhanced lipid peroxidation. VHL is a major tumour suppressor in ccRCC and loss of VHL leads to stabilisation of hypoxia inducible factors HIF-1α and HIF-2α. Restoration of functional VHL via exogenous expression of pVHL reverted ccRCC cells to an oxidative metabolism and rendered them insensitive to the induction of ferroptosis. VHL reconstituted cells also exhibited reduced lipid storage and higher expression of genes associated with oxidiative phosphorylation and fatty acid metabolism. Importantly, inhibition of β-oxidation or mitochondrial ATP-synthesis restored ferroptosis sensitivity in VHL reconstituted cells. We also found that inhibition of GSH synthesis blocked tumour growth in a MYC-dependent mouse model of renal cancer. Together, our data suggest that reduced fatty acid metabolism due to inhibition of β-oxidation renders renal cancer cells highly dependent on the GSH/GPX pathway to prevent lipid peroxidation and ferroptotic cell death.

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Additional information

These authors contributed equally: Heike Miess, Beatrice Dankworth.

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Acknowledgements

We thank the LRI research services for technical support. We also thank Prof. William Kaelin (Dana Faber Cancer Institute, Boston) for providing isogenic renal cancer cell lines. This study was funded by Cancer Research UK, the German Research Foundation (FOR2314) and the German Cancer Aid (111917).

Author contributions

HM and AS conceived the project and wrote the manuscript. HM performed the starvation and rescue experiments, the RNAi screen and most of the silencing experiments and cell analyses. BD performed ferroptosis analyses including rescue experiments and RNA analyses. W.S. performed metabolite analyses. BP contributed to the study design and manuscript preparation. MJ, BS and MH were involved in optimisation, execution and analysis of the RNAi screen. MR performed histological analysis and interpretation. AG and DF provided the mouse model and performed the in vivo experiments. All authors commented on the manuscript.

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    Affiliations

    1. Gene Expression Analysis Laboratory, Cancer Research UK London Research Institute, 44 Lincoln’s Inn Fields, London, WC2A 3LY, UK

      • Heike Miess
      • , Barrie Peck
      •  & Almut Schulze
    2. Oncogene Biology Laboratory, The Francis Crick Institute, 1 Midland Road London, London, NW1 1AT, UK

      • Heike Miess
      •  & Julian Downward
    3. Theodor-Boveri-Institute, Biocenter, Am Hubland, 97074, Würzburg, Germany

      • Beatrice Dankworth
      • , Werner Schmitz
      •  & Almut Schulze
    4. Division of Medical Oncology, Stanford University School of Medicine, Stanford, CA, 94305, USA

      • Arvin M. Gouw
      •  & Dean W. Felsher
    5. Institute of Pathology, University Hospital Würzburg, 97080, Würzburg, Germany

      • Mathias Rosenfeldt
    6. High Throughput Screening, The Francis Crick Institute, 1 Midland Road London, London, NW1 1AT, UK

      • Ming Jiang
      • , Becky Saunders
      •  & Michael Howell
    7. Division of Cancer Biology, The Institute of Cancer Research, 237 Fulham Road, London, SW7 3RP, UK

      • Julian Downward
      •  & Barrie Peck
    8. Comprehensive Cancer Center Mainfranken, Josef-Schneider-Str.6, 97080, Würzburg, Germany

      • Almut Schulze

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    Correspondence to Almut Schulze.

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    DOI

    https://doi.org/10.1038/s41388-018-0315-z