Tyrosine phosphatase SHP2 promotes breast cancer progression and maintains tumor-initiating cells via activation of key transcription factors and a positive feedback signaling loop

Abstract

New cancer therapies are likely to arise from an in-depth understanding of the signaling networks influencing tumor initiation, progression and metastasis. We show a fundamental role for Src-homology 2 domain-containing phosphatase 2 (SHP2) in these processes in human epidermal growth factor receptor 2 (HER2)-positive and triple-negative breast cancers. Knockdown of SHP2 eradicated breast tumor-initiating cells in xenograft models, and SHP2 depletion also prevented invasion in three-dimensional cultures and in a transductal invasion assay in vivo. Notably, SHP2 knockdown in established breast tumors blocked their growth and reduced metastasis. Mechanistically, SHP2 activated stemness-associated transcription factors, including v-myc myelocytomatosis viral oncogene homolog (c-Myc) and zinc finger E-box binding homeobox 1 (ZEB1), which resulted in the repression of let-7 microRNA and the expression of a set of 'SHP2 signature' genes. We found these genes to be simultaneously activated in a large subset of human primary breast tumors that are associated with invasive behavior and poor prognosis. These results provide new insights into the signaling cascades influencing tumor-initiating cells as well as a rationale for targeting SHP2 in breast cancer.

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Figure 1: SHP2 promotes invasion, proliferation and loss of polarity in three-dimensional cultures and in vivo.
Figure 2: SHP2 promotes tumor maintenance, growth and progression to lung metastases.
Figure 3: SHP2 maintains TICs in vivo.
Figure 4: SHP2 acts through the ERK pathway in breast cancer.
Figure 5: SHP2 increases the activity of key transcription factors and represses let-7 miRNA.
Figure 6: SHP2 is active in a large subset of breast tumors associated with poor prognosis.

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Acknowledgements

We thank N. Hynes (FMI) and G. Christofori (University of Basel) for helpful comments on the manuscript, S. Gasser, W. Filipowicz, H. Grosshans, J. Krol (FMI) and R.B. Clarke (Manchester University) for helpful discussions, W. Xian (in the lab of J. Brugge, Harvard Medical School) and D. Medina (Baylor College of Medicine) for help with the intraductal protocol, T. Westbrook (Baylor College of Medicine) for providing the dox-inducible lentiviral vector, C. Kuperwasser (Tufts University) for providing the SUM159 and SUM1315 cells, V. Reddy (GNF) for providing the primary breast tumor, and A. Terrier (Novartis Institutes for Biomedical Research) for excellent technical assistance, as well as additional members of the Bentires-Alj lab for advice and discussions and various colleagues for reagents. We also thank the International Genomics Consortium and expO (http://www.intgen.org/expo/) for offering free access to clinically annotated tumor data. Research in the lab of M.B.-A. is supported by the Novartis Research Foundation, the European Research Council (ERC starting grant 243211-PTPsBDC), the Swiss Cancer League and the Krebsliga Beider Basel.

Author information

N.A. and M.B.-A. designed and performed most of the experiments and wrote the manuscript. N.S. and H.B. performed experiments related to the biochemical mechanism of action SHP2, real-time PCR and lung metastases experiments. D.G. and M.B.S. performed bioinformatic analyses. G.M.-B. performed the RPA. M.Q., G.M. and S.C. performed the tissue microarray and examined histological sections from the xenografts. P.J.B., M.P. and E.v.N. performed the motif activity response analysis. G.H. and S.J.E. generated the dox-inducible lentiviral vector. All the authors participated in the preparation of the manuscript.

Correspondence to Mohamed Bentires-Alj.

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Aceto, N., Sausgruber, N., Brinkhaus, H. et al. Tyrosine phosphatase SHP2 promotes breast cancer progression and maintains tumor-initiating cells via activation of key transcription factors and a positive feedback signaling loop. Nat Med 18, 529–537 (2012). https://doi.org/10.1038/nm.2645

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