Abstract

Tumours exist in a hypoxic microenvironment and must limit excessive oxygen consumption. Hypoxia-inducible factor (HIF) controls mitochondrial oxygen consumption, but how/if tumours regulate non-mitochondrial oxygen consumption (NMOC) is unknown. Protein-tyrosine phosphatase-1B (PTP1B) is required for Her2/Neu-driven breast cancer (BC) in mice, although the underlying mechanism and human relevance remain unclear. We found that PTP1B-deficient HER2+ xenografts have increased hypoxia, necrosis and impaired growth. In vitro, PTP1B deficiency sensitizes HER2+ BC lines to hypoxia by increasing NMOC by α-KG-dependent dioxygenases (α-KGDDs). The moyamoya disease gene product RNF213, an E3 ligase, is negatively regulated by PTP1B in HER2+ BC cells. RNF213 knockdown reverses the effects of PTP1B deficiency on α-KGDDs, NMOC and hypoxia-induced death of HER2+ BC cells, and partially restores tumorigenicity. We conclude that PTP1B acts via RNF213 to suppress α-KGDD activity and NMOC. This PTP1B/RNF213/α-KGDD pathway is critical for survival of HER2+ BC, and possibly other malignancies, in the hypoxic tumour microenvironment.

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Acknowledgements

We thank G. Keller and T. W. Mak (Princess Margaret Cancer Center) for helpful comments on the manuscript. This work was funded by NIH grant R37 CA49152 and Canadian Institutes of Health Research (CIHR) grant 120593 (to B.G.N.), CIHR grant 62975 (to J.W.D.), CIHR grant 136956 (to S.S.S.), CIHR grant 133615 (to T.K.), Terry Fox New Frontiers Research Program PPG09-02005 (to B.G.W.), Cancer Research-UK and the Wellcome Trust (to S.E.W. and C.J.S), NIH grant GM96745 (to S.P.G.) and Kiban Kenkyu grant A-25253047 to A.K. Work in the Neel and Wouters laboratories was partially supported by the Princess Margaret Cancer Foundation and the Ontario Ministry of Health and Long Term Care. B.G.N. and J.W.D. are Canada Research Chairs, Tier 1, and B.G.W. is a Senior Investigator of the Ontario Institute for Cancer Research. T.K. is supported by the Canada Research Chair programme (Tier 2). R.M. was partially supported by a Post-doctoral Fellowship Grant, and R.S.B. by a Doctoral Fellowship Grant, both from the Canadian Breast Cancer Foundation. W.Z. was supported by a CIHR Post-doctoral Fellowship Grant. D.C. was supported by an Ontario Graduate Scholarship. A.A.R. was supported by a MITACS-Accelerate internship. A.S. was supported by the Medical Biophysics Excellence Award and the Kirsti Piia Callum Memorial Fellowship.

Author information

Author notes

    • Caterina Iorio
    • , Richard Marcotte
    •  & Yang Xu

    These authors contributed equally to this work.

Affiliations

  1. Department of Medical Biophysics, University of Toronto, Toronto, Ontario M5G 2M9, Canada

    • Robert S. Banh
    • , Yang Xu
    • , Dan Cojocari
    • , Ankit Sinha
    • , Ronald Wu
    • , Thomas Kislinger
    •  & Bradly G. Wouters
  2. Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario M5G 1L7, Canada

    • Robert S. Banh
    • , Caterina Iorio
    • , Richard Marcotte
    • , Yang Xu
    • , Dan Cojocari
    • , Ankit Sinha
    • , Ronald Wu
    • , Carl Virtanen
    • , Thomas Kislinger
    • , Bradly G. Wouters
    •  & Benjamin G. Neel
  3. Laura and Isaac Perlmutter Cancer Center, New York University Langone Medical Center, New York University, New York, New York 10016, USA

    • Robert S. Banh
    • , Yang Xu
    • , Shuang Zhang
    •  & Benjamin G. Neel
  4. Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Ontario M5G 1X5, Canada

    • Anas Abdel Rahman
    • , Judy Pawling
    •  & James W. Dennis
  5. Department of Genetics, Research Center, King Faisal Specialist Hospital and Research Center, Riyadh 12713, Saudi Arabia

    • Anas Abdel Rahman
  6. Donnelly Centre for Cellular and Biomolecular Research, Banting and Best Department of Medical Research, University of Toronto, Toronto, Ontario M5S 3E1, Canada

    • Wei Zhang
    •  & Sachdev S. Sidhu
  7. Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA

    • Christopher M. Rose
    • , Marta Isasa
    •  & Steven P. Gygi
  8. Department of Health and Environmental Sciences, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan

    • Toshiaki Hitomi
    •  & Akio Koizumi
  9. Department of Radiation System Biology, Institute of Radiation Biology Center, Kyoto University, Kyoto 606-8501, Japan

    • Toshiyuki Habu
  10. Chemistry Research Laboratory, Oxford University, 12 Mansfield Road, Oxford OX1 3TA, UK

    • Sarah E. Wilkins
    •  & Christopher J. Schofield

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Contributions

R.S.B. designed and performed most of the experiments, analysed and interpreted the data and wrote the manuscript. C.I. performed and analysed in vitro cell growth and tumour growth experiments. Y.X. performed LC–MS/MS to identify PTP1B-interacting proteins and substrates. R.M. provided conceptual advice and helped to design experiments. D.C. helped set up oxygen consumption measurements. A.A.R., J.P. and J.W.D. prepared, performed and helped to analyse the metabolomics experiments. W.Z. and S.S.S. assisted with the auto-ubiquitylation assays. A.S. performed LC–MS/MS to identify ubiquitylated proteins from HA–Ub pulldowns. C.M.R. and M.I. performed anti-diGly IP–MS to identify endogenous ubiquitylated peptides. S.Z. generated the RNF213-KO line and assisted with some of the ubiquitylation experiments. R.W. performed dot blots with anti-5-meC antibodies. C.V. helped with the bioinformatic analyses. T.Hitomi, T.Habu and A.K. provided reagents for RNF213 detection and expression. S.E.W. and C.J.S. provided conceptual advice and reagents for α-KGDDs. B.G.W. provided conceptual advice on hypoxia and metabolism experiments. B.G.N. conceived and supervised the project, helped to interpret the data and wrote the manuscript. All authors critically analysed data, and edited and approved the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Benjamin G. Neel.

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https://doi.org/10.1038/ncb3376

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