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Inhibition of MAPK pathway is essential for suppressing Rheb-Y35N driven tumor growth

Oncogene volume 36, pages 756–765 (2017)Cite this article

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Abstract

Rheb is a Ras family GTPase, which binds to and activates mammalian target of rapamycin complex 1 (mTORC1) when GTP loaded. Recently, cancer genome sequencing efforts have identified recurrent Rheb Tyr35Asn mutations in kidney and endometrial carcinoma. Here we show that Rheb-Y35N causes not only constitutive mTORC1 activation, but sustained activation of the MEK-ERK pathway in a TSC1/TSC2/TBC1D7 protein complex and mTORC1-independent manner, contributing to intrinsic resistance to rapamycin. Rheb-Y35N transforms NIH3T3 cells, resulting in aggressive tumor formation in xenograft nude mice, which could be suppressed by combined treatment with rapamycin and an extracellular signal-regulated kinase (ERK) inhibitor. Furthermore, Rheb-Y35N inhibits AMPKα activation in response to nutrient depletion or 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR), leading to attenuated phosphorylation of BRAF-S729 and retained mitogen-activated protein kinase (MAPK) activation. Finally, we demonstrate that Rheb-WT can bind AMPK to facilitate AMPK activation, whereas Rheb-Y35N competitively binds AMPK, impairing AMPK phosphorylation. In summary, our findings indicate that Rheb-Y35N is a dominantly active tumor driver that activates both mTORC1 and MAPK to promote tumor growth, suggesting a combination of mTORC1 and MAPK inhibitors may be of therapeutic value in patients whose cancers sustain this mutation.

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Acknowledgements

This study was supported by NIH NCI 1P01CA120964. We thank Nathanael Gray, Heidi Greulich and John Blenis for the gift of reagents.

Author contributions

YW performed experiments and analyzed data. XFH, JW and YLY assisted with the immunoblotanalysis and in vivo mice experiment. YZ, ZL and LHZ designed and analyzed quantification proteomic data. YBZ and GHL performed the structural protein analysis. H-LP and GWX contribute to the experiment design and edits. YL and DJK designed and guided all experiments, interpreted data and wrote the manuscript.

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Authors and Affiliations

  1. Department of Biotechnology, Scientific Research Center for Translational Medicine, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China

    Y Wang, Y Yin, H-l Piao & Y Liu

  2. School of life Science, Dalian University, Dalian, China

    Y Wang & J Wang

  3. Department of General Surgery, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China

    X Hong

  4. Department of Biotechnology, Laboratory of Molecular Modeling and Design, State Key Laboratory of Molecular Reaction Dynamics, Dalian Institute of Chemical Physics, Chinese Academy of Sciences Dalian, Liaoning, China

    Y Zhang & G Li

  5. Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China

    Y Zhou, H-l Piao, Z Liang, L Zhang, G Xu & Y Liu

  6. Divisions of Pulmonary and Critical Care Medicine, Brigham & Women Hospital, Harvard Medical School, Boston, MA, USA

    D J Kwiatkowski & Y Liu

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  1. Y Wang
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Correspondence to D J Kwiatkowski or Y Liu.

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Wang, Y., Hong, X., Wang, J. et al. Inhibition of MAPK pathway is essential for suppressing Rheb-Y35N driven tumor growth. Oncogene 36, 756–765 (2017). https://doi.org/10.1038/onc.2016.246

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  • DOI: https://doi.org/10.1038/onc.2016.246

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