Abstract

Approximately 200 BRAF mutant alleles have been identified in human tumours. Activating BRAF mutants cause feedback inhibition of GTP-bound RAS, are RAS-independent and signal either as active monomers (class 1) or constitutively active dimers (class 2)1. Here we characterize a third class of BRAF mutants—those that have impaired kinase activity or are kinase-dead. These mutants are sensitive to ERK-mediated feedback and their activation of signalling is RAS-dependent. The mutants bind more tightly than wild-type BRAF to RAS–GTP, and their binding to and activation of wild-type CRAF is enhanced, leading to increased ERK signalling. The model suggests that dysregulation of signalling by these mutants in tumours requires coexistent mechanisms for maintaining RAS activation despite ERK-dependent feedback. Consistent with this hypothesis, melanomas with these class 3 BRAF mutations also harbour RAS mutations or NF1 deletions. By contrast, in lung and colorectal cancers with class 3 BRAF mutants, RAS is typically activated by receptor tyrosine kinase signalling. These tumours are sensitive to the inhibition of RAS activation by inhibitors of receptor tyrosine kinases. We have thus defined three distinct functional classes of BRAF mutants in human tumours. The mutants activate ERK signalling by different mechanisms that dictate their sensitivity to therapeutic inhibitors of the pathway.

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Acknowledgements

We are grateful to P. Lito, Y. Gao, W. Su, L. Desrochers, D. Santamaria and O. Abdel-Wahab for useful discussions. We thank S. Lowe for the vectors of the retrovirus-based inducible expression system and M. Baccarini for the Raf1-knockout MEFs. We thank Novartis for supplying INC280. We also thank I. Maruani Ryan for her help on this work. This research was supported by grants to N.R. from the National Institutes of Health (NIH) (P01 CA129243; R35 CA210085); the Melanoma Research Alliance (237059 and 348724); The Commonwealth Foundation for Cancer Research and The Center for Experimental Therapeutics at Memorial Sloan Kettering Cancer Center; and the Stand Up To Cancer – American Cancer Society Lung Cancer Dream Team Translational Research Grant (SU2C-AACR-DT17-15). Support was also received from the NIH MSKCC Cancer Center Support Grant P30 CA008748. Additional funding was provided by a Career Development Award from the Conquer Cancer Foundation of the American Society of Clinical Oncology (R.Y.); and from the NIH (R01 CA204749 and R01 CA180037), the Sontag and Josie Robertson Foundations and the Cycle For Survival (B.S.T.). Additional NIH funding was received by E.d.S. (U54 ODSS020355). We would like to acknowledge the support of the Arlene and Joseph Taub Foundation and of Paula and Thomas McInerney, without which this work would not have been possible. M.T.C. was supported in part by the NIH training grant T32 GM007175. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. This work was also supported by grants from the European Research Council (ERC-AG/250297-RAS AHEAD), EU-Framework Programme (HEALTH-F2-2010-259770/LUNGTARGET and HEALTH-2010-260791/EUROCANPLATFORM) and Spanish Ministry of Economy and Competitiveness (SAF2011-30173 and SAF2014-59864-R) to M.B. M.B. is the recipient of an Endowed Chair from the AXA Research Fund.

Author information

Affiliations

  1. Program in Molecular Pharmacology, Memorial Sloan Kettering Cancer Center, New York, New York 10065, USA

    • Zhan Yao
    • , Vanessa S. Rodrik-Outmezguine
    • , Neilawattie M. Torres
    • , Huiyong Zhao
    • , Linde Miles
    • , Elisa de Stanchina
    •  & Neal Rosen
  2. Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, New York 10065, USA

    • Rona Yaeger
    • , David B. Solit
    •  & Neal Rosen
  3. Center for Neural Science, College of Arts and Sciences, New York University, New York, New York 10012, USA

    • Anthony Tao
  4. Human Oncology and Pathogenesis Program, Memorial Sloan Kettering Cancer Center, New York, New York 10065, USA

    • Matthew T. Chang
    • , David B. Solit
    •  & Barry S. Taylor
  5. Department of Epidemiology and Biostatistics, Memorial Sloan Kettering Cancer Center, New York, New York 10065, USA

    • Matthew T. Chang
    •  & Barry S. Taylor
  6. Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, California 94158, USA

    • Matthew T. Chang
  7. Molecular Oncology Programme, Centro Nacional de Investigaciones Oncológicas (CNIO), Melchor Fernández Almagro 3, Madrid 28029, Spain

    • Matthias Drosten
    •  & Mariano Barbacid
  8. Molecular Oncology Group, NantOmics, LLC, 9600 Medical Center Drive, Suite 300, Rockville, Maryland 20854, USA

    • Fabiola Cecchi
    •  & Todd Hembrough
  9. Département de médecine oncologique, Gustave Roussy Cancer Campus, Villejuif, France

    • Judith Michels
  10. Faculté de médecine, Université Paris Sud, Le Kremlin-Bicêtre, France

    • Judith Michels
  11. Urology Department, Saint Joseph Hospital, Paris, France

    • Hervé Baumert
  12. Anti-Cancer Drug Development Graduate Training Program, Department of Pharmacology and Molecular Sciences, Johns Hopkins University, Baltimore, Maryland 21205, USA

    • Linde Miles
  13. Department of Radiology, Memorial Sloan Kettering Cancer Center, New York, New York 10065, USA

    • Naomi M. Campbell
  14. Center for Molecular Oncology, Memorial Sloan Kettering Cancer Center, New York, New York 10065, USA

    • David B. Solit
    •  & Barry S. Taylor
  15. Center for Mechanism-Based Therapeutics, Memorial Sloan Kettering Cancer Center, New York, New York 10065, USA

    • Neal Rosen

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Contributions

Z.Y., R.Y., B.S.T. and N.R. conceived the project, designed the experiments. Z.Y. and N.R. wrote the manuscript. R.Y., D.B.S., V.S.R.-O., B.S.T. and M.B. provided critical revisions of the manuscript. Z.Y., R.Y., A.T., N.M.T., M.D., H.Z., V.S.R.-O., F.C., T.H., L.M., E.D.S., J.M., H.B. and M.B. established the in vitro and in vivo experimental systems, performed the laboratory experiments and analysed the results. M.T.C., D.B.S. and B.S.T. acquired and analysed the genetic data. N.M.C. reviewed and interpreted the clinical computerized axial tomography scans.

Competing interests

N.R. is on the scientific advisory board of and receives research funding from Chugai, and is on the scientific advisory board of and owns stock in Beigene, Wellspring and Kura. N.R. is also on the scientific advisory board of Daiichi-Sankyo, Astra-Zeneca and Takeda, and is a consultant to Novartis. V.S.R.-O. is now an employee of Novartis Pharmaceutical Corporation as Precision Medicine Associate Director. F.C. is an employee at NantOmics, LLC. T.H. is the President of Proteomics at NantOmics and owns the stock.

Corresponding author

Correspondence to Neal Rosen.

Reviewer Information Nature thanks L. Garraway and the other anonymous reviewer(s) for their contribution to the peer review of this work.

Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Extended data

Supplementary information

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  1. 1.

    Supplementary Figures

    Supplementary Figures 1-11 containing the uncropped blots from Figures 1-3 and Extended Data Figures 1-4.

  2. 2.

    Supplementary Information

    This file contains Supplementary Discussion parts 1 and 2, Supplementary Figures S1-S3, and Oligos for mutagenesis of Class 3 BRAF mutants

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DOI

https://doi.org/10.1038/nature23291

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