Abstract

We identified the Rho GTPase Rnd1 as a candidate metastasis suppressor in basal-like and triple-negative breast cancer through bioinformatics analysis. Depletion of Rnd1 disrupted epithelial adhesion and polarity, induced epithelial-to-mesenchymal transition, and cooperated with deregulated expression of c-Myc or loss of p53 to cause neoplastic conversion. Mechanistic studies revealed that Rnd1 suppresses Ras signalling by activating the GAP domain of Plexin B1, which inhibits Rap1. Rap1 inhibition in turn led to derepression of p120 Ras-GAP, which was able to inhibit Ras. Inactivation of Rnd1 in mammary epithelial cells induced highly undifferentiated and invasive tumours in mice. Conversely, Rnd1 expression inhibited spontaneous and experimental lung colonization in mouse models of metastasis. Genomic studies indicated that gene deletion in combination with epigenetic silencing or, more rarely, point mutation inactivates RND1 in human breast cancer. These results reveal a previously unappreciated mechanism through which Rnd1 restrains activation of Ras-MAPK signalling and breast tumour initiation and progression.

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Acknowledgements

We thank X. Zhang for advice, reagents, and for sharing the structure of the Plexin-GAP–Rap1 complex before its publication, R. A. Weinberg, J. S. Brugge, M. Overholtzer, A. Ridley, B. Park, L. Tamagnome, M. Resh, H. Djaballah, T. Kataoka, M. Negishi and D. Medina for reagents, K. Manova for assistance with confocal microscopy, R. Khanin and G. P. Gupta for help with multivariate analysis, M. Buck for comments on the manuscript, and members of the Giancotti laboratory for discussions. We thank the Geoffrey Beene Translational Oncology Core, the Genomic Core, and the HTG Core of MSKCC for technical assistance. This work was supported by grants from the National Institutes of Health (P01 CA094060 Project 4 to F.G.G. and P30 CA08748 to MSKCC) and the Geoffrey Beene Cancer Center at MSKCC.

Author information

Author notes

    • Tomoyo Okada
    • , Gaia Schiavon
    • , Miguel A. López-Lago
    • , Christine A. Pratilas
    • , Adriana Heguy
    •  & Giorgio Inghirami

    Present addresses: Department of Surgery, Memorial Sloan-Kettering Cancer Center, New York 10065, USA (T.O.); Breast Unit, Royal Marsden Hospital, London SW3 6JJ, UK (G.S.); Department of Surgery and Center for Cell Engineering, Memorial Sloan-Kettering Cancer Center, New York 10065, USA (M.A.L-L.); The Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, Maryland 21287, USA (C.A.P.); Department of Pathology, Office for Collaborative Science, New York University, New York 10016, USA (A.H.); Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York 10065, USA (G.I.).

    • Tomoyo Okada
    • , Surajit Sinha
    •  & Ilaria Esposito

    These authors contributed equally to this work.

Affiliations

  1. Cell Biology Program, Sloan-Kettering Institute for Cancer Research, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA

    • Tomoyo Okada
    • , Surajit Sinha
    • , Ilaria Esposito
    • , Gaia Schiavon
    • , Miguel A. López-Lago
    • , Wenjing Su
    • , Jonathan M. Hernandez
    •  & Filippo G. Giancotti
  2. Molecular Pharmacology and Chemistry Program, Sloan-Kettering Institute for Cancer Research, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA

    • Christine A. Pratilas
    •  & Neal Rosen
  3. Department of Pediatrics, Memorial Hospital, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA

    • Christine A. Pratilas
  4. Department of Biomedical Sciences and Human Oncology, Center of Experimental Medicine and Research, University of Torino, Torino 10126, Italy

    • Cristina Abele
    •  & Giorgio Inghirami
  5. Department of Surgery, Memorial Hospital, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA

    • Jonathan M. Hernandez
  6. Department of Surgical Oncology, Research Institute for Radiation Biology and Medicine, Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima 734-8551, Japan

    • Masahiro Ohara
    •  & Morihito Okada
  7. Genomics Core Facility, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA

    • Agnes Viale
  8. Geoffrey Beene Translational Oncology Core Facility, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA

    • Adriana Heguy
  9. Bioinformatics Core Facility, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA

    • Nicholas D. Socci
  10. Department of Medical Sciences, Center of Experimental Medicine and Research, University of Torino, Torino 10126, Italy

    • Anna Sapino
  11. Department of Epidemiology and Biostatistics, Memorial Hospital, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA

    • Venkatraman E. Seshan
  12. Structural Biology Program, Sloan-Kettering Institute for Cancer Research, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA

    • Stephen Long
  13. Department of Medicine, Memorial Hospital, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA

    • Neal Rosen

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Contributions

T.O., S.S., I.E., G.S., M.A.L-L., W.S., C.A.P., J.M.H. and C.A. performed and interpreted experiments; M.Ohara, M.Okada and A.S. provided annotated breast tumour samples, A.V. designed and performed genomic analyses; G.S. and V.E.S. performed bioinformatic analyses; S.L. provided structural insight; G.I. examined the results of FISH experiments; N.R. and G.I. supervised some of the experiments; F.G.G. supervised the entire study and wrote the paper.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Filippo G. Giancotti.

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https://doi.org/10.1038/ncb3082

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