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Profiling of residual breast cancers after neoadjuvant chemotherapy identifies DUSP4 deficiency as a mechanism of drug resistance

Nature Medicine volume 18, pages 1052–1059 (2012)Cite this article

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Abstract

Neoadjuvant chemotherapy (NAC) induces a pathological complete response (pCR) in ∼30% of patients with breast cancer. However, many patients have residual cancer after chemotherapy, which correlates with a higher risk of metastatic recurrence and poorer outcome than those who achieve a pCR. We hypothesized that molecular profiling of tumors after NAC would identify genes associated with drug resistance. Digital transcript counting was used to profile surgically resected breast cancers after NAC. Low concentrations of dual specificity protein phosphatase 4 (DUSP4), an ERK phosphatase, correlated with high post-NAC tumor cell proliferation and with basal-like breast cancer (BLBC) status. BLBC had higher DUSP4 promoter methylation and gene expression patterns of Ras-ERK pathway activation relative to other breast cancer subtypes. DUSP4 overexpression increased chemotherapy-induced apoptosis, whereas DUSP4 depletion dampened the response to chemotherapy. Reduced DUSP4 expression in primary tumors after NAC was associated with treatment-refractory high Ki-67 scores and shorter recurrence-free survival. Finally, inhibition of mitogen-activated protein kinase kinase (MEK) synergized with docetaxel treatment in BLBC xenografts. Thus, DUSP4 downregulation activates the Ras-ERK pathway in BLBC, resulting in an attenuated response to anti-cancer chemotherapy.

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Figure 1: Ki-67–associated gene expression in chemotherapy-refractory breast cancers.
Figure 2: DUSP4 expression is deficient in BLBC and associates with Ras-ERK pathway activation.
Figure 3: DUSP4 expression is inversely associated with Ras-ERK pathway activation.
Figure 4: DUSP4 regulates MAPK signaling and cell viability.
Figure 5: Reduction of DUSP4 expression after NAC correlates with enhanced tumor Ki-67 score and shorter RFS.
Figure 6: Pharmacological or genetic inhibition of MEK improves chemotherapy-induced apoptosis in vitro and in vivo.

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Gene Expression Omnibus

NCBI Reference Sequence

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Acknowledgements

We would like to thank E. Penni Black (University of Kentucky), who provided the AdMEK1ca adenoviral construct. The authors would also like to thank P.D. Smith and AstraZeneca for supplying selumetinib utilized in the in vivo experiments. This work was supported by Breast Cancer Specialized Program of Research Excellence (SPORE) grant P50CA98131, Vanderbilt-Ingram Cancer Center Support grant P30CA68485, a grant from the Breast Cancer Research Foundation, the American Cancer Society Clinical Research Professorship grant CRP-07-234 and the Lee Jeans Translational Breast Cancer Research Program (to C.L.A.). G.B.M. was supported by the SU2C Dream Team award, PO1CA099031 and the Komen Promise grant KG 081694. J.S. and M.D. were funded by the Breakthrough Breast Cancer and the Royal Marsden National Institutes of Health Research Biomedical Research Centre.

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Authors and Affiliations

  1. Department of Medicine, Vanderbilt-Ingram Comprehensive Cancer Center, Vanderbilt University, Nashville, Tennessee, USA

    Justin M Balko, Neil E Bhola & Carlos L Arteaga

  2. Department of Cancer Biology, Vanderbilt-Ingram Comprehensive Cancer Center, Vanderbilt University, Nashville, Tennessee, USA

    Rebecca S Cook, David B Vaught, Todd W Miller & Carlos L Arteaga

  3. Breast Cancer Research Program, Vanderbilt-Ingram Comprehensive Cancer Center, Vanderbilt University, Nashville, Tennessee, USA

    Rebecca S Cook, Todd W Miller, Melinda E Sanders, Ingrid M Meszoely & Carlos L Arteaga

  4. Department of Pathology, Vanderbilt-Ingram Comprehensive Cancer Center, Vanderbilt University, Nashville, Tennessee, USA

    María G Kuba, Melinda E Sanders & Nara M Granja-Ingram

  5. Department of Surgery, Vanderbilt-Ingram Comprehensive Cancer Center, Vanderbilt University, Nashville, Tennessee, USA

    J Joshua Smith & Ingrid M Meszoely

  6. Breakthrough Breast Cancer Centre, Institute of Cancer Research, Royal Marsden Hospital, London, UK

    Janine Salter & Mitch Dowsett

  7. Academic Department of Biochemistry, Royal Marsden Hospital, London, UK

    Janine Salter & Mitch Dowsett

  8. Department of Systems Biology, University of Texas, M.D. Anderson Cancer Center, Houston, Texas, USA

    Katherine Stemke-Hale, Ana M González-Angulo & Gordon B Mills

  9. Department of Breast Medical Oncology, University of Texas, M.D. Anderson Cancer Center, Houston, Texas, USA

    Ana M González-Angulo

  10. División de Investigación, Oncosalud, Lima, Perú

    Joseph A Pinto

  11. Instituto Nacional de Enfermedades Neoplásicas, Lima, Perú

    Henry L Gómez

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Contributions

J.M.B. designed and performed the experiments, and authored the manuscript. R.S.C. provided expert guidance on in vivo models and performed immunofluorescence. M.G.K., N.M.G.-I. and M.E.S. provided pathology support and performed IHC staining and scoring. D.B.V., T.W.M. and N.E.B. provided scientific advice. K.S.-H., A.M.G.-A. and G.B.M. performed RPPA analysis. J.S., M.D., J.A.P. and H.L.G. provided post-NAC patient tumor samples. J.J.S. and I.M.M. identified the Vanderbilt cohort and managed the clinical database, including performing retrospective chart review. M.E.S. and I.M.M. functioned as honest brokers to maintain patient privacy. C.L.A. provided scientific direction, established collaborations, prepared the manuscript with J.M.B. and allocated funding for the work. All the authors participated in the preparation of the manuscript.

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Correspondence to Carlos L Arteaga.

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Competing interests

M.D. receives research grant funds and has received honoraria for consulting and advisory board work from AstraZeneca.

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Balko, J., Cook, R., Vaught, D. et al. Profiling of residual breast cancers after neoadjuvant chemotherapy identifies DUSP4 deficiency as a mechanism of drug resistance. Nat Med 18, 1052–1059 (2012). https://doi.org/10.1038/nm.2795

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