Hsp90 inhibition suppresses mutant EGFR-T790M signaling and overcomes kinase inhibitor resistance.

The epidermal growth factor receptor (EGFR) secondary kinase domain T790M non-small cell lung cancer (NSCLC) mutation enhances receptor catalytic activity and confers resistance to the reversible tyrosine kinase inhibitors gefitinib and erlotinib. Currently, irreversible inhibitors represent the primary approach in clinical use to circumvent resistance. We show that higher concentrations of the irreversible EGFR inhibitor CL-387,785 are required to inhibit EGFR phosphorylation in T790M-expressing cells compared with EGFR mutant NSCLC cells without T790M. Additionally, CL-387,785 does not fully suppress phosphorylation of other activated receptor tyrosine kinases (RTK) in T790M-expressing cells. These deficiencies result in residual Akt and mammalian target of rapamycin (mTOR) activities. Full suppression of EGFR-mediated signaling in T790M-expressing cells requires the combination of CL-387,785 and rapamycin. In contrast, Hsp90 inhibition overcomes these limitations in vitro and depletes cells of EGFR, other RTKs, and phospho-Akt and inhibits mTOR signaling whether or not T790M is present. EGFR-T790M-expressing cells rendered resistant to CL-387,785 by a kinase switch mechanism retain sensitivity to Hsp90 inhibition. Finally, Hsp90 inhibition causes regression in murine lung adenocarcinomas driven by mutant EGFR (L858R) with or without T790M. However, efficacy in the L858R-T790M model requires a more intense treatment schedule and responses were transient. Nonetheless, these findings suggest that Hsp90 inhibitors may be effective in T790M-expressing cells and offer an alternative therapeutic strategy for this subset of lung cancers.

[1]  M. Meyerson,et al.  The T790M mutation in EGFR kinase causes drug resistance by increasing the affinity for ATP , 2008, Proceedings of the National Academy of Sciences.

[2]  N. Hanna,et al.  MET Amplification Leads to Gefitinib Resistance in Lung Cancer by Activating ERBB3 Signaling , 2008 .

[3]  William Pao,et al.  MET amplification occurs with or without T790M mutations in EGFR mutant lung tumors with acquired resistance to gefitinib or erlotinib , 2007, Proceedings of the National Academy of Sciences.

[4]  W. Pao,et al.  Development of New Mouse Lung Tumor Models Expressing EGFR T790M Mutants Associated with Clinical Resistance to Kinase Inhibitors , 2007, PloS one.

[5]  L. Neckers,et al.  Sensitivity of epidermal growth factor receptor and ErbB2 exon 20 insertion mutants to Hsp90 inhibition , 2007, British Journal of Cancer.

[6]  M. Meyerson,et al.  Bronchial and peripheral murine lung carcinomas induced by T790M-L858R mutant EGFR respond to HKI-272 and rapamycin combination therapy. , 2007, Cancer cell.

[7]  David M Sabatini,et al.  Defining the role of mTOR in cancer. , 2007, Cancer cell.

[8]  H. Garewal,et al.  A novel tyrosine kinase switch is a mechanism of imatinib resistance in gastrointestinal stromal tumors , 2007, Oncogene.

[9]  M. Meyerson,et al.  Allele-dependent variation in the relative cellular potency of distinct EGFR inhibitors , 2007, Cancer biology & therapy.

[10]  Paul Workman,et al.  Gene and protein expression profiling of human ovarian cancer cells treated with the heat shock protein 90 inhibitor 17-allylamino-17-demethoxygeldanamycin. , 2007, Cancer research.

[11]  Matthew Meyerson,et al.  Structures of lung cancer-derived EGFR mutants and inhibitor complexes: mechanism of activation and insights into differential inhibitor sensitivity. , 2007, Cancer cell.

[12]  D. Haber,et al.  Epidermal growth factor receptor mutants from human lung cancers exhibit enhanced catalytic activity and increased sensitivity to gefitinib. , 2007, Cancer research.

[13]  Daniel A. Haber,et al.  Epidermal growth factor receptor mutations in lung cancer , 2007, Nature Reviews Cancer.

[14]  H. Ji,et al.  Therapeutic anti-EGFR antibody 806 generates responses in murine de novo EGFR mutant-dependent lung carcinomas. , 2007, The Journal of clinical investigation.

[15]  T. Golub,et al.  Transcriptional profiling identifies cyclin D1 as a critical downstream effector of mutant epidermal growth factor receptor signaling. , 2006, Cancer research.

[16]  Edward S. Kim,et al.  Heterodimerization of insulin-like growth factor receptor/epidermal growth factor receptor and induction of survivin expression counteract the antitumor action of erlotinib. , 2006, Cancer research.

[17]  N. Rosen,et al.  Association with HSP90 inhibits Cbl-mediated down-regulation of mutant epidermal growth factor receptors. , 2006, Cancer research.

[18]  T. Lynch,et al.  HKI-272, an irreversible pan erbB receptor tyrosine kinase inhibitor: Preliminary phase 1 results in patients with solid tumors. , 2006, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[19]  D. Bottaro,et al.  Targeting the c-Met Signaling Pathway in Cancer , 2006, Clinical Cancer Research.

[20]  P. Jänne,et al.  The impact of human EGFR kinase domain mutations on lung tumorigenesis and in vivo sensitivity to EGFR-targeted therapies. , 2006, Cancer cell.

[21]  Manuel Morente,et al.  Molecular Context of the EGFR Mutations: Evidence for the Activation of mTOR/S6K Signaling , 2006, Clinical Cancer Research.

[22]  Gordon B Mills,et al.  mTOR inhibition induces upstream receptor tyrosine kinase signaling and activates Akt. , 2006, Cancer research.

[23]  N. Rosen,et al.  V600E B-Raf requires the Hsp90 chaperone for stability and is degraded in response to Hsp90 inhibitors. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[24]  Paul Workman,et al.  Inhibitors of the HSP90 molecular chaperone: current status. , 2006, Advances in cancer research.

[25]  P. Workman,et al.  Hsp90 inhibitors in the clinic. , 2006, Handbook of experimental pharmacology.

[26]  A. Nakashima,et al.  Suppression of the mTOR-raptor signaling pathway by the inhibitor of heat shock protein 90 geldanamycin. , 2006, Journal of biochemistry.

[27]  P. Workman,et al.  Activated B-RAF is an Hsp90 client protein that is targeted by the anticancer drug 17-allylamino-17-demethoxygeldanamycin. , 2005, Cancer research.

[28]  M. Meyerson,et al.  An alternative inhibitor overcomes resistance caused by a mutation of the epidermal growth factor receptor. , 2005, Cancer research.

[29]  F. Khuri,et al.  Activation of Akt and eIF4E survival pathways by rapamycin-mediated mammalian target of rapamycin inhibition. , 2005, Cancer research.

[30]  William Pao,et al.  Inhibition of drug-resistant mutants of ABL, KIT, and EGF receptor kinases. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[31]  A. Lowell,et al.  Epidermal growth factor receptors harboring kinase domain mutations associate with the heat shock protein 90 chaperone and are destabilized following exposure to geldanamycins. , 2005, Cancer research.

[32]  Patricia L. Harris,et al.  Irreversible inhibitors of the EGF receptor may circumvent acquired resistance to gefitinib. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[33]  C. Mermel,et al.  ErbB-3 mediates phosphoinositide 3-kinase activity in gefitinib-sensitive non-small cell lung cancer cell lines. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[34]  M. Meyerson,et al.  EGFR mutation and resistance of non-small-cell lung cancer to gefitinib. , 2005, The New England journal of medicine.

[35]  H. Varmus,et al.  Acquired Resistance of Lung Adenocarcinomas to Gefitinib or Erlotinib Is Associated with a Second Mutation in the EGFR Kinase Domain , 2005, PLoS medicine.

[36]  Ramaswamy Nilakantan,et al.  Optimization of 6,7-disubstituted-4-(arylamino)quinoline-3-carbonitriles as orally active, irreversible inhibitors of human epidermal growth factor receptor-2 kinase activity. , 2005, Journal of medicinal chemistry.

[37]  S. Zlotkin,et al.  Micronutrient Sprinkles to Control Childhood Anaemia , 2005, PLoS medicine.

[38]  Yaoquan Liu,et al.  Synthesis and biological activities of novel 17-aminogeldanamycin derivatives. , 2004, Bioorganic & medicinal chemistry.

[39]  Daniel A. Haber,et al.  Gefitinib-Sensitizing EGFR Mutations in Lung Cancer Activate Anti-Apoptotic Pathways , 2004, Science.

[40]  T. Nielsen,et al.  Expression of the Insulin-Like Growth Factor I Receptor and Urokinase Plasminogen Activator in Breast Cancer Is Associated with Poor Survival , 2004, Cancer Research.

[41]  E. Sausville,et al.  Clinical development of 17-allylamino, 17-demethoxygeldanamycin. , 2003, Current cancer drug targets.

[42]  N. Rosen,et al.  Akt Forms an Intracellular Complex with Heat Shock Protein 90 (Hsp90) and Cdc37 and Is Destabilized by Inhibitors of Hsp90 Function* , 2002, The Journal of Biological Chemistry.

[43]  I. Weinstein.  Addiction to Oncogenes--the Achilles Heal of Cancer , 2002, Science.

[44]  P. Workman,et al.  Inhibition of signal transduction by the Hsp90 inhibitor 17-allylamino-17-demethoxygeldanamycin results in cytostasis and apoptosis. , 2001, Cancer research.

[45]  Y. Yarden,et al.  Sensitivity of Mature ErbB2 to Geldanamycin Is Conferred by Its Kinase Domain and Is Mediated by the Chaperone Protein Hsp90* , 2001, The Journal of Biological Chemistry.

[46]  T. Tsuruo,et al.  Modulation of Akt kinase activity by binding to Hsp90. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[47]  E. Sausville,et al.  The geldanamycins are potent inhibitors of the hepatocyte growth factor/scatter factor-met-urokinase plasminogen activator-plasmin proteolytic network. , 2000, Cancer research.

[48]  D. Kitchen,et al.  Irreversible inhibition of epidermal growth factor receptor tyrosine kinase with in vivo activity by N-[4-[(3-bromophenyl)amino]-6-quinazolinyl]-2-butynamide (CL-387,785). , 1999, Biochemical pharmacology.