Activated Phosphoinositide 3-Kinase/AKT Signaling Confers Resistance to Trastuzumab but not Lapatinib

Trastuzumab and lapatinib provide clinical benefit to women with human epidermal growth factor receptor 2 (HER)–positive breast cancer. However, not all patients whose tumors contain the HER2 alteration respond. Consequently, there is an urgent need to identify new predictive factors for these agents. The aim of this study was to investigate the role of receptor tyrosine kinase signaling and phosphoinositide 3-kinase (PI3K)/AKT pathway activation in conferring resistance to trastuzumab and lapatinib. To address this question, we evaluated response to trastuzumab and lapatinib in a panel of 18 HER2-amplified cell lines, using both two- and three-dimensional culture. The SUM-225, HCC-1419, HCC-1954, UACC-893, HCC-1569, UACC-732, JIMT-1, and MDA-453 cell lines were found to be innately resistant to trastuzumab, whereas the MDA-361, MDA-453, HCC-1569, UACC-732, JIMT-1, HCC-202, and UACC-893 cells are innately lapatinib resistant. Lapatinib was active in de novo (SUM-225, HCC-1419, and HCC-1954) and in a BT-474 cell line with acquired resistance to trastuzumab. In these cells, trastuzumab had little effect on AKT phosphorylation, whereas lapatinib retained activity through the dephosphorylation of AKT. Increased phosphorylation of HER2, epidermal growth factor receptor, HER3, and insulin-like growth factor IR correlated with response to lapatinib but not trastuzumab. Loss of PTEN or the presence of activating mutations in PI3K marked resistance to trastuzumab, but lapatinib response was independent of these factors. Thus, increased activation of the PI3K/AKT pathway correlates with resistance to trastuzumab, which can be overcome by lapatinib. In conclusion, pharmacologic targeting of the PI3K/AKT pathway may provide benefit to HER2-positive breast cancer patients who are resistant to trastuzumab therapy. Mol Cancer Ther; 9(6); 1489–502. ©2010 AACR.

[1]  S. Kane,et al.  Differential sensitivities of trastuzumab (Herceptin®)-resistant human breast cancer cells to phosphoinositide-3 kinase (PI-3K) and epidermal growth factor receptor (EGFR) kinase inhibitors , 2005, Breast Cancer Research and Treatment.

[2]  R. Nahta,et al.  Lapatinib induces apoptosis in trastuzumab-resistant breast cancer cells: effects on insulin-like growth factor I signaling , 2007, Molecular Cancer Therapeutics.

[3]  Joe W. Gray,et al.  HER2 signaling pathway activation and response of breast cancer cells to HER2-targeting agents is dependent strongly on the 3D microenvironment , 2010, Breast Cancer Research and Treatment.

[4]  H. Lane,et al.  Modulation of p27/Cdk2 complex formation through 4D5-mediated inhibition of HER2 receptor signaling. , 2001, Annals of oncology : official journal of the European Society for Medical Oncology.

[5]  T. Fleming,et al.  Use of chemotherapy plus a monoclonal antibody against HER2 for metastatic breast cancer that overexpresses HER2. , 2001, The New England journal of medicine.

[6]  Violeta Serra,et al.  Phosphatidylinositol 3-kinase hyperactivation results in lapatinib resistance that is reversed by the mTOR/phosphatidylinositol 3-kinase inhibitor NVP-BEZ235. , 2008, Cancer research.

[7]  F. Montemurro,et al.  Trastuzumab-based combination therapy for breast cancer , 2004, Expert opinion on pharmacotherapy.

[8]  M. Sliwkowski,et al.  A central role for HER3 in HER2-amplified breast cancer: implications for targeted therapy. , 2008, Cancer research.

[9]  C. Tacchetti,et al.  TGFα expression impairs Trastuzumab-induced HER2 downregulation , 2005, Oncogene.

[10]  J. Fricker.  San Antonio Breast Cancer Symposium. , 2009, The Lancet. Oncology.

[11]  J. Baselga,et al.  Updated Survival Analysis of a Randomized Study of Lapatinib Alone or in Combination with Trastuzumab in Women with HER2-Positive Metastatic Breast Cancer Progressing on Trastuzumab Therapy. , 2009 .

[12]  M. Arbushites,et al.  HER-2 Gene Amplification, HER-2 and Epidermal Growth Factor Receptor mRNA and Protein Expression, and Lapatinib Efficacy in Women with Metastatic Breast Cancer , 2008, Clinical Cancer Research.

[13]  K. Lackey,et al.  The effects of the novel, reversible epidermal growth factor receptor/ErbB-2 tyrosine kinase inhibitor, GW2016, on the growth of human normal and tumor-derived cell lines in vitro and in vivo. , 2001, Molecular cancer therapeutics.

[14]  N. Robert,et al.  Multinational study of the efficacy and safety of humanized anti-HER2 monoclonal antibody in women who have HER2-overexpressing metastatic breast cancer that has progressed after chemotherapy for metastatic disease. , 1999, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[15]  Suzanne F. Jones,et al.  Phase I safety, pharmacokinetics, and clinical activity study of lapatinib (GW572016), a reversible dual inhibitor of epidermal growth factor receptor tyrosine kinases, in heavily pretreated patients with metastatic carcinomas. , 2005, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[16]  J. Engelman,et al.  Breast cancer-associated PIK3CA mutations are oncogenic in mammary epithelial cells. , 2005, Cancer research.

[17]  Ming Tan,et al.  PTEN activation contributes to tumor inhibition by trastuzumab, and loss of PTEN predicts trastuzumab resistance in patients. , 2004, Cancer cell.

[18]  J. Baselga,et al.  Randomized study of Lapatinib alone or in combination with trastuzumab in women with ErbB2-positive, trastuzumab-refractory metastatic breast cancer. , 2010, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[19]  S. Stein,et al.  Lapatinib antitumor activity is not dependent upon phosphatase and tensin homologue deleted on chromosome 10 in ErbB2-overexpressing breast cancers. , 2007, Cancer research.

[20]  G. Mills,et al.  Preclinical Testing of Clinically Applicable Strategies for Overcoming Trastuzumab Resistance Caused by PTEN Deficiency , 2007, Clinical Cancer Research.

[21]  T. Mukohara,et al.  Association between gain-of-function mutations in PIK3CA and resistance to HER2-targeted agents in HER2-amplified breast cancer cell lines. , 2010, Annals of oncology : official journal of the European Society for Medical Oncology.

[22]  Brian Higgins,et al.  Targeting ligand-activated ErbB2 signaling inhibits breast and prostate tumor growth. , 2002, Cancer cell.

[23]  J. Baselga,et al.  Expression of p95HER2, a truncated form of the HER2 receptor, and response to anti-HER2 therapies in breast cancer. , 2007, Journal of the National Cancer Institute.

[24]  Y. Lu,et al.  Insulin-like growth factor-I receptor signaling and resistance to trastuzumab (Herceptin). , 2001, Journal of the National Cancer Institute.

[25]  P. Hegde,et al.  A model of acquired autoresistance to a potent ErbB2 tyrosine kinase inhibitor and a therapeutic strategy to prevent its onset in breast cancer , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[26]  M. Sliwkowski,et al.  Ligand-independent HER2/HER3/PI3K complex is disrupted by trastuzumab and is effectively inhibited by the PI3K inhibitor GDC-0941. , 2009, Cancer cell.

[27]  F. Bertucci,et al.  ERBB2 phosphorylation and trastuzumab sensitivity of breast cancer cell lines , 2007, Oncogene.

[28]  Lyndsay N Harris,et al.  Efficacy and safety of trastuzumab as a single agent in first-line treatment of HER2-overexpressing metastatic breast cancer. , 2002, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[29]  C. Ries,et al.  Comparison of 3D and 2D tumor models reveals enhanced HER2 activation in 3D associated with an increased response to trastuzumab , 2009, Oncogene.

[30]  Hanina Hibshoosh,et al.  PIK3CA mutations correlate with hormone receptors, node metastasis, and ERBB2, and are mutually exclusive with PTEN loss in human breast carcinoma. , 2005, Cancer research.

[31]  W Godolphin,et al.  Studies of the HER-2/neu proto-oncogene in human breast and ovarian cancer. , 1989, Science.

[32]  D. Slamon,et al.  Transformation mediated by the human HER-2 gene independent of the epidermal growth factor receptor. , 1992, Oncogene.

[33]  P. LoRusso,et al.  Phase II study of predictive biomarker profiles for response targeting human epidermal growth factor receptor 2 (HER-2) in advanced inflammatory breast cancer with lapatinib monotherapy. , 2008, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[34]  J. Baselga,et al.  A randomized study of lapatinib alone or in combination with trastuzumab in heavily pretreated HER2+ metastatic breast cancer progressing on trastuzumab therapy , 2008 .

[35]  C R King,et al.  erbB-2 is a potent oncogene when overexpressed in NIH/3T3 cells. , 1987, Science.

[36]  R. Schilsky,et al.  Oral capecitabine vs intravenous 5-fluorouracil and leucovorin: integrated efficacy data and novel analyses from two large, randomised, phase III trials , 2004, British Journal of Cancer.

[37]  J. Crown,et al.  HER-2 signaling and inhibition in breast cancer. , 2009, Current cancer drug targets.

[38]  H. Drexler,et al.  Detection of mycoplasma in leukemia–lymphoma cell lines using polymerase chain reaction , 2002, Leukemia.

[39]  B. Leyland-Jones,et al.  Recombinant human insulin-like growth factor binding protein 3 inhibits growth of human epidermal growth factor receptor-2-overexpressing breast tumors and potentiates herceptin activity in vivo. , 2006, Cancer research.

[40]  J. Ptak,et al.  High Frequency of Mutations of the PIK3CA Gene in Human Cancers , 2004, Science.

[41]  M. Kraus,et al.  Efficient coupling with phosphatidylinositol 3-kinase, but not phospholipase C gamma or GTPase-activating protein, distinguishes ErbB-3 signaling from that of other ErbB/EGFR family members , 1994, Molecular and cellular biology.

[42]  M. Berger,et al.  Activity of the dual kinase inhibitor lapatinib (GW572016) against HER-2-overexpressing and trastuzumab-treated breast cancer cells. , 2006, Cancer research.

[43]  F. Maurer,et al.  The ErbB2/ErbB3 heterodimer functions as an oncogenic unit: ErbB2 requires ErbB3 to drive breast tumor cell proliferation , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[44]  P. Depowski,et al.  Loss of Expression of the PTEN Gene Protein Product Is Associated with Poor Outcome in Breast Cancer , 2001, Modern Pathology.

[45]  M. Berger,et al.  Lapatinib plus capecitabine for HER2-positive advanced breast cancer. , 2006, The New England journal of medicine.

[46]  A. M. Stanley,et al.  Structure of the extracellular region of HER 2 alone and in complex with the Herceptin Fab , 2022 .

[47]  Charles Ginther,et al.  Identifying individuals by sequencing mitochondrial DNA from teeth , 1992, Nature Genetics.

[48]  Joel Greshock,et al.  Novel mechanism of lapatinib resistance in HER2-positive breast tumor cells: activation of AXL. , 2009, Cancer research.

[49]  W. McGuire,et al.  Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene. , 1987, Science.

[50]  G. Hortobagyi,et al.  The efficacy of Herceptin therapies is influenced by the expression of other erbB receptors, their ligands and the activation of downstream signalling proteins , 2004, British Journal of Cancer.

[51]  Carlos L Arteaga,et al.  Human Breast Cancer Cells Selected for Resistance to Trastuzumab In vivo Overexpress Epidermal Growth Factor Receptor and ErbB Ligands and Remain Dependent on the ErbB Receptor Network , 2007, Clinical Cancer Research.

[52]  Ryuji Kobayashi,et al.  Insulin-like growth factor-I receptor/human epidermal growth factor receptor 2 heterodimerization contributes to trastuzumab resistance of breast cancer cells. , 2005, Cancer research.