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Regulation of endocytosis via the oxygen-sensing pathway

Nature Medicine volume 15, pages 319–324 (2009)Cite this article

Abstract

Tumor hypoxia is associated with disease progression, resistance to conventional cancer therapies and poor prognosis1,2. Hypoxia, by largely unknown mechanisms, leads to deregulated accumulation of and signaling via receptor tyrosine kinases (RTKs) that are critical for driving oncogenesis. Here, we show that hypoxia or loss of von Hippel–Lindau protein—the principal negative regulator of hypoxia-inducible factor (HIF)3—prolongs the activation of epidermal growth factor receptor that is attributable to lengthened receptor half-life and retention in the endocytic pathway. The deceleration in endocytosis is due to the attenuation of Rab5-mediated early endosome fusion via HIF-dependent downregulation of a critical Rab5 effector, rabaptin-5, at the level of transcription. Primary kidney and breast tumors with strong hypoxic signatures show significantly lower expression of rabaptin-5 RNA and protein. These findings reveal a general role of the oxygen-sensing pathway in endocytosis and support a model in which tumor hypoxia or oncogenic activation of HIF prolongs RTK-mediated signaling by delaying endocytosis-mediated deactivation of receptors.

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Figure 1: Loss of VHL prolongs turnover and downstream signaling of EGFR in a HIF-dependent manner.
Figure 2: Regulation of Rab5-mediated endosome fusion is HIF dependent and influences the clearance rate of internalized EGF.
Figure 3: Hypoxia or loss of VHL attenuates the expression of the Rab5 effector rabaptin-5.
Figure 4: Expression of rabaptin-5 is sufficient to modulate early endosome fusion and is inversely associated with the physical recruitment of HIF to the HRE on RABEP1 promoter.

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Gene Expression Omnibus

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Acknowledgements

This work was supported by grants from the Canadian Institutes of Health Research (CIHR MOP77718 to M.O., MOP37778 to P.A.M. and CIHR team grant to M.P.). Y.W. and O.R. are recipients of CIHR postdoctoral fellowships. M.S.I. and M.O. are Canada Research Chairs.

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Authors and Affiliations

  1. Department of Laboratory Medicine and Pathobiology, St. Michael's Hospital, University of Toronto, 1 King's College Circle, Toronto, M5S 1A8, Ontario, Canada

    Yi Wang, Olga Roche, Andrew J Evans, Julie L Metcalf, Meredith S Irwin, Philip A Marsden & Michael Ohh

  2. Department of Medical Biophysics, St. Michael's Hospital, University of Toronto, 1 King's College Circle, Toronto, M5S 1A8, Ontario, Canada

    Mathew S Yan & Philip A Marsden

  3. Department of Medicine, St. Michael's Hospital, University of Toronto, 1 King's College Circle, Toronto, M5S 1A8, Ontario, Canada

    Mathew S Yan

  4. Department of Biochemistry, McGill University, 687 Pine Ave. West, Montreal, H3A 1A1, Quebec, Canada

    Greg Finak & Morag Park

  5. Department of Pathology, University Health Network, Princess Margaret Hospital, 610 University Ave., Toronto, M5G 2M9, Ontario, Canada

    Andrew J Evans

  6. Department of Haematology Oncology, The Lineberger Comprehensive Cancer Centre, 102 Mason Farm Rd., CB7295, University of North Carolina, Chapel Hill, 27599, North Carolina, USA

    Bridgid E Hast, Sara C Hanna & William Y Kim

  7. Van Andel Research Institute, 333 Bostwick Ave. NE, Grand Rapids, 49503, Michigan, USA

    Bill Wondergem, Kyle A Furge & Bin T Teh

  8. Department of Paediatrics, Hospital for Sick Children and University of Toronto, 555 University Ave., Toronto, M5G 1X8, Ontario, Canada

    Meredith S Irwin

  9. Division of Cell Biology, Hospital for Sick Children and University of Toronto, 555 University Ave., Toronto, M5G 1X8, Ontario, Canada

    Sergio Grinstein

  10. Department of Biochemistry, Hospital for Sick Children and University of Toronto, 555 University Ave., Toronto, M5G 1X8, Ontario, Canada

    Sergio Grinstein

  11. Department of Oncology, McGill University, 687 Pine Ave. West, Montreal, H3A 1A1, Quebec, Canada

    Morag Park

  12. Department of Medicine, McGill University, 687 Pine Ave. West, Montreal, H3A 1A1, Quebec, Canada

    Morag Park

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Contributions

Y.W. designed and performed experiments and wrote the manuscript. O.R. performed EMSA. M.S.Y. performed ChIP analysis. G.F. performed immunofluorescence and mRNA expression analyses of breast tumor samples. A.J.E. performed CCRCC immunohistology. J.L.M performed western blot analysis of CCRCC samples. B.E.H. and S.C.H. performed fluorescence microscopy of MEFs. B.W. and K.A.F. performed western blot and mRNA expression analyses of CCRCC samples. M.S.I., W.Y.K, B.T.T., S.G., M.P. and P.A.M. contributed conceptually to the project and manuscript. M.O. directed the project, designed the experiments and wrote the manuscript.

Corresponding author

Correspondence to Michael Ohh.

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Supplementary Figures 1–13, Supplementary Results, Supplementary Methods and Supplementary References (PDF 8943 kb)

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Wang, Y., Roche, O., Yan, M. et al. Regulation of endocytosis via the oxygen-sensing pathway. Nat Med 15, 319–324 (2009). https://doi.org/10.1038/nm.1922

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