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Phosphoproteomic mass spectrometry profiling links Src family kinases to escape from HER2 tyrosine kinase inhibition

Oncogene volume 30, pages 4163–4174 (2011)Cite this article

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Abstract

Despite the initial effectiveness of the tyrosine kinase inhibitor lapatinib against HER2 gene-amplified breast cancers, most patients eventually relapse after treatment, implying that tumors acquire mechanisms of drug resistance. To discover these mechanisms, we generated six lapatinib-resistant HER2-overexpressing human breast cancer cell lines. In cells that grew in the presence of lapatinib, HER2 autophosphorylation was undetectable, whereas active phosphoinositide-3 kinase (PI3K)-Akt and mitogen-activated protein kinase (MAPK) were maintained. To identify networks maintaining these signaling pathways, we profiled the tyrosine phosphoproteome of sensitive and resistant cells using an immunoaffinity-enriched mass spectrometry method. We found increased phosphorylation of Src family kinases (SFKs) and putative Src substrates in several resistant cell lines. Treatment of these resistant cells with Src kinase inhibitors partially blocked PI3K-Akt signaling and restored lapatinib sensitivity. Further, SFK mRNA expression was upregulated in primary HER2+ tumors treated with lapatinib. Finally, the combination of lapatinib and the Src inhibitor AZD0530 was more effective than lapatinib alone at inhibiting pAkt and growth of established HER2-positive BT-474 xenografts in athymic mice. These data suggest that increased Src kinase activity is a mechanism of lapatinib resistance and support the combination of HER2 antagonists with Src inhibitors early in the treatment of HER2+ breast cancers in order to prevent or overcome resistance to HER2 inhibitors.

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Acknowledgements

This work was supported by NIH R01 CA80195 (CLA), Dinah Armstrong Kukes Fund/Phi Mu foundation (BNR), T32 CA119910 (BNR), Department of Defense BC087465 Post-Doctoral Fellowship (BNR), ASCO Young Investigator Award (BNR), Kleberg Center for Molecular Markers at MD Anderson Cancer Center, ASCO Career Development Award (AMG), NCI 1K23CA121994-01 (AMG), Susan G Komen Foundation FAS0703849 (AMG, GBM), ACS Clinical Research Professorship CRP-07-234 (CLA), Lee Jeans Translational Breast Cancer Research Program (CLA), Breast Cancer SPORE P50 CA98131 and Vanderbilt-Ingram Cancer Center Support Grant P30 CA68485.

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Authors and Affiliations

  1. Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN, USA

    B N Rexer, C Rinehart & C L Arteaga

  2. Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN, USA

    A-J L Ham, S Hill & D C Liebler

  3. Ayers Institute of Precancer Detection and Diagnosis, Vanderbilt University School of Medicine, Nashville, TN, USA

    A-J L Ham & D C Liebler

  4. Department of Pathology, Vanderbilt University School of Medicine, Nashville, TN, USA

    N de Matos Granja-Ingram

  5. Department of Systems Biology, MD Anderson Cancer Center, Houston, TX, USA

    A M González-Angulo & G B Mills

  6. Department of Breast Medical Oncology, MD Anderson Cancer Center, Houston, TX, USA

    A M González-Angulo

  7. Department of Gynecologic Medical Oncology, MD Anderson Cancer Center, Houston, TX, USA

    G B Mills

  8. Lester and Sue Smith Breast Center, Baylor College of Medicine, Houston, TX, USA

    B Dave

  9. Department of Medicine, The Methodist Hospital, Houston, TX, USA

    J C Chang

  10. Department of Cancer Biology, Vanderbilt University School of Medicine, Nashville, TN, USA

    C L Arteaga

  11. Department of Breast Cancer Research Program, Vanderbilt-Ingram Cancer Center, Nashville, TN, USA

    C L Arteaga

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  1. B N Rexer
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Rexer, B., Ham, AJ., Rinehart, C. et al. Phosphoproteomic mass spectrometry profiling links Src family kinases to escape from HER2 tyrosine kinase inhibition. Oncogene 30, 4163–4174 (2011). https://doi.org/10.1038/onc.2011.130

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