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01-08-2016 | PRECLINICAL STUDIES

AZD9291 overcomes T790 M-mediated resistance through degradation of EGFR^L858R/T790M in non-small cell lung cancer cells

Authors: Bo Mi Ku, Yeon-Hee Bae, Jiae Koh, Jong-Mu Sun, Se-hoon Lee, Jin Seok Ahn, Keunchil Park, Myung-Ju Ahn

Published in: Investigational New Drugs | Issue 4/2016

Summary

The discovery of activating mutations of epidermal growth factor receptor (EGFR) has resulted in the development of more effective treatments for non-small cell lung cancer (NSCLC). Although first-generation EGFR tyrosine kinase inhibitors (EGFR TKIs) provide significant clinical benefit, acquired resistance often occurs, most commonly (>50 %) via a T790 M resistance mutation. Although AZD9291 is selective for both T790 M and activating EGFR mutations over wild-type EGFR, it is highly active when T790 M is present, especially EGFR^L858R/T790M, and modestly active when T790 M is absent. The aim of this study was to elucidate the underlying mechanism of the high sensitivity of NSCLC cells harboring EGFR^L858R/T790M to AZD9291. In H1975 cells harboring EGFR^L858R/T790M, AZD9291 potently inhibited cellular growth and EGFR signaling pathways together with depletion of mutant EGFR protein. AZD9291-induced depletion of EGFR^L858R/T790M protein was abrogated through inhibition of the proteasome with MG132. However, AZD9291 had no effect on protein levels of EGFR^WT and EGFR^L858R. In addition, AZD9291 induced apoptosis and caused expression changes in cell cycle-related genes. Moreover, oral administration of AZD9291 as a single agent induced tumor regression in vivo in a H1975 tumor xenograft model and reduced EGFR^L858R/T790M protein levels in xenograft tumors. Taken together, our results provide a potential mechanism for the sensitivity of EGFR^L858R/T790M cells to AZD9291 and suggest that AZD9291 may be effective in cases of T790 M-positive EGFR resistance.

Cataldo VD, Gibbons DL, Perez-Soler R, Quintas-Cardama A (2011) Treatment of non-small-cell lung cancer with erlotinib or gefitinib. N Engl J Med 364(10):947–955. doi:10.1056/NEJMct0807960 CrossRefPubMed

Engelman JA, Janne PA (2008) Mechanisms of acquired resistance to epidermal growth factor receptor tyrosine kinase inhibitors in non-small cell lung cancer. Clinical cancer research: an official journal of the American Association for Cancer Research 14(10):2895–2899. doi:10.1158/1078-0432.CCR-07-2248 CrossRef

Pao W, Chmielecki J (2010) Rational, biologically based treatment of EGFR-mutant non-small-cell lung cancer. Nat Rev Cancer 10(11):760–774. doi:10.1038/nrc2947 CrossRefPubMedPubMedCentral

Camidge DR, Pao W, Sequist LV (2014) Acquired resistance to TKIs in solid tumours: learning from lung cancer. Nature reviews Clinical oncology 11(8):473–481. doi:10.1038/nrclinonc.2014.104 CrossRefPubMed

Gainor JF, Shaw AT (2013) Emerging paradigms in the development of resistance to tyrosine kinase inhibitors in lung cancer. J Clin Oncol Off J Am Soc Clin Oncol 31(31):3987–3996. doi:10.1200/JCO.2012.45.2029 CrossRef

Remon J, Moran T, Majem M, Reguart N, Dalmau E, Marquez-Medina D, Lianes P (2014) Acquired resistance to epidermal growth factor receptor tyrosine kinase inhibitors in EGFR-mutant non-small cell lung cancer: a new era begins. Cancer Treat Rev 40(1):93–101. doi:10.1016/j.ctrv.2013.06.002 CrossRefPubMed

Sos ML, Rode HB, Heynck S, Peifer M, Fischer F, Kluter S, Pawar VG, Reuter C, Heuckmann JM, Weiss J, Ruddigkeit L, Rabiller M, Koker M, Simard JR, Getlik M, Yuza Y, Chen TH, Greulich H, Thomas RK, Rauh D (2010) Chemogenomic profiling provides insights into the limited activity of irreversible EGFR inhibitors in tumor cells expressing the T790 M EGFR resistance mutation. Cancer Res 70(3):868–874. doi:10.1158/0008-5472.CAN-09-3106 CrossRefPubMed

Pao W, Miller VA, Politi KA, Riely GJ, Somwar R, Zakowski MF, Kris MG, Varmus H (2005) Acquired resistance of lung adenocarcinomas to gefitinib or erlotinib is associated with a second mutation in the EGFR kinase domain. PLoS Med 2(3):e73. doi:10.1371/journal.pmed.0020073 CrossRefPubMedPubMedCentral

Yun CH, Mengwasser KE, Toms AV, Woo MS, Greulich H, Wong KK, Meyerson M, Eck MJ (2008) The T790 M mutation in EGFR kinase causes drug resistance by increasing the affinity for ATP. Proc Natl Acad Sci U S A 105(6):2070–2075. doi:10.1073/pnas.0709662105 CrossRefPubMedPubMedCentral

10.

Chong CR, Janne PA (2013) The quest to overcome resistance to EGFR-targeted therapies in cancer. Nat Med 19(11):1389–1400. doi:10.1038/nm.3388 CrossRefPubMedPubMedCentral

11.

Li D, Ambrogio L, Shimamura T, Kubo S, Takahashi M, Chirieac LR, Padera RF, Shapiro GI, Baum A, Himmelsbach F, Rettig WJ, Meyerson M, Solca F, Greulich H, Wong KK (2008) BIBW2992, an irreversible EGFR/HER2 inhibitor highly effective in preclinical lung cancer models. Oncogene 27(34):4702–4711. doi:10.1038/onc.2008.109 CrossRefPubMedPubMedCentral

12.

Engelman JA, Zejnullahu K, Gale CM, Lifshits E, Gonzales AJ, Shimamura T, Zhao F, Vincent PW, Naumov GN, Bradner JE, Althaus IW, Gandhi L, Shapiro GI, Nelson JM, Heymach JV, Meyerson M, Wong KK, Janne PA (2007) PF00299804, an irreversible pan-ERBB inhibitor, is effective in lung cancer models with EGFR and ERBB2 mutations that are resistant to gefitinib. Cancer Res 67(24):11924–11932. doi:10.1158/0008-5472.CAN-07-1885 CrossRefPubMed

13.

Zhou W, Ercan D, Chen L, Yun CH, Li D, Capelletti M, Cortot AB, Chirieac L, Iacob RE, Padera R, Engen JR, Wong KK, Eck MJ, Gray NS, Janne PA (2009) Novel mutant-selective EGFR kinase inhibitors against EGFR T790 M. Nature 462(7276):1070–1074. doi:10.1038/nature08622 CrossRefPubMedPubMedCentral

14.

Miller VA, Hirsh V, Cadranel J, Chen YM, Park K, Kim SW, Zhou C, Su WC, Wang M, Sun Y, Heo DS, Crino L, Tan EH, Chao TY, Shahidi M, Cong XJ, Lorence RM, Yang JC (2012) Afatinib versus placebo for patients with advanced, metastatic non-small-cell lung cancer after failure of erlotinib, gefitinib, or both, and one or two lines of chemotherapy (LUX-Lung 1): a phase 2b/3 randomised trial. The lancet oncology 13(5):528–538. doi:10.1016/S1470-2045(12)70087-6 CrossRefPubMed

15.

Kim Y, Ko J, Cui Z, Abolhoda A, Ahn JS, Ou SH, Ahn MJ, Park K (2012) The EGFR T790 M mutation in acquired resistance to an irreversible second-generation EGFR inhibitor. Mol Cancer Ther 11(3):784–791. doi:10.1158/1535-7163.MCT-11-0750 CrossRefPubMed

16.

Finlay MR, Anderton M, Ashton S, Ballard P, Bethel PA, Box MR, Bradbury RH, Brown SJ, Butterworth S, Campbell A, Chorley C, Colclough N, Cross DA, Currie GS, Grist M, Hassall L, Hill GB, James D, James M, Kemmitt P, Klinowska T, Lamont G, Lamont SG, Martin N, McFarland HL, Mellor MJ, Orme JP, Perkins D, Perkins P, Richmond G, Smith P, Ward RA, Waring MJ, Whittaker D, Wells S, Wrigley GL (2014) Discovery of a potent and selective EGFR inhibitor (AZD9291) of both sensitizing and T790 M resistance mutations that spares the wild type form of the receptor. J Med Chem 57(20):8249–8267. doi:10.1021/jm500973a CrossRefPubMed

17.

Walter AO, Sjin RT, Haringsma HJ, Ohashi K, Sun J, Lee K, Dubrovskiy A, Labenski M, Zhu Z, Wang Z, Sheets M, St Martin T, Karp R, van Kalken D, Chaturvedi P, Niu D, Nacht M, Petter RC, Westlin W, Lin K, Jaw-Tsai S, Raponi M, Van Dyke T, Etter J, Weaver Z, Pao W, Singh J, Simmons AD, Harding TC, Allen A (2013) Discovery of a mutant-selective covalent inhibitor of EGFR that overcomes T790 M-mediated resistance in NSCLC. Cancer discovery 3(12):1404–1415. doi:10.1158/2159-8290.CD-13-0314 CrossRefPubMedPubMedCentral

18.

Peters S, Zimmermann S, Adjei AA (2014) Oral epidermal growth factor receptor tyrosine kinase inhibitors for the treatment of non-small cell lung cancer: comparative pharmacokinetics and drug-drug interactions. Cancer Treat Rev 40(8):917–926. doi:10.1016/j.ctrv.2014.06.010 CrossRefPubMed

19.

Cross DA, Ashton SE, Ghiorghiu S, Eberlein C, Nebhan CA, Spitzler PJ, Orme JP, Finlay MR, Ward RA, Mellor MJ, Hughes G, Rahi A, Jacobs VN, Red Brewer M, Ichihara E, Sun J, Jin H, Ballard P, Al-Kadhimi K, Rowlinson R, Klinowska T, Richmond GH, Cantarini M, Kim DW, Ranson MR, Pao W (2014) AZD9291, an irreversible EGFR TKI, overcomes T790 M-mediated resistance to EGFR inhibitors in lung cancer. Cancer discovery 4(9):1046–1061. doi:10.1158/2159-8290.CD-14-0337 CrossRefPubMedPubMedCentral

20.

Janne PA, Yang JC, Kim DW, Planchard D, Ohe Y, Ramalingam SS, Ahn MJ, Kim SW, Su WC, Horn L, Haggstrom D, Felip E, Kim JH, Frewer P, Cantarini M, Brown KH, Dickinson PA, Ghiorghiu S, Ranson M (2015) AZD9291 in EGFR inhibitor-resistant non-small-cell lung cancer. N Engl J Med 372(18):1689–1699. doi:10.1056/NEJMoa1411817 CrossRefPubMed

21.

Tomas A, Futter CE, Eden ER (2014) EGF receptor trafficking: consequences for signaling and cancer. Trends Cell Biol 24(1):26–34. doi:10.1016/j.tcb.2013.11.002 CrossRefPubMedPubMedCentral

22.

Chung BM, Tom E, Zutshi N, Bielecki TA, Band V, Band H (2014) Nexus of signaling and endocytosis in oncogenesis driven by non-small cell lung cancer-associated epidermal growth factor receptor mutants. World journal of clinical oncology 5(5):806–823. doi:10.5306/wjco.v5.i5.806 CrossRefPubMedPubMedCentral

23.

Shtiegman K, Kochupurakkal BS, Zwang Y, Pines G, Starr A, Vexler A, Citri A, Katz M, Lavi S, Ben-Basat Y, Benjamin S, Corso S, Gan J, Yosef RB, Giordano S, Yarden Y (2007) Defective ubiquitinylation of EGFR mutants of lung cancer confers prolonged signaling. Oncogene 26(49):6968–6978. doi:10.1038/sj.onc.1210503 CrossRefPubMed

24.

Red Brewer M, Yun CH, Lai D, Lemmon MA, Eck MJ, Pao W (2013) Mechanism for activation of mutated epidermal growth factor receptors in lung cancer. Proc Natl Acad Sci U S A 110(38):E3595–E3604. doi:10.1073/pnas.1220050110 CrossRefPubMedPubMedCentral

25.

Sawai A, Chandarlapaty S, Greulich H, Gonen M, Ye Q, Arteaga CL, Sellers W, Rosen N, Solit DB (2008) Inhibition of Hsp90 down-regulates mutant epidermal growth factor receptor (EGFR) expression and sensitizes EGFR mutant tumors to paclitaxel. Cancer Res 68(2):589–596. doi:10.1158/0008-5472.CAN-07-1570 CrossRefPubMedPubMedCentral

26.

Shimamura T, Li D, Ji H, Haringsma HJ, Liniker E, Borgman CL, Lowell AM, Minami Y, McNamara K, Perera SA, Zaghlul S, Thomas RK, Greulich H, Kobayashi S, Chirieac LR, Padera RF, Kubo S, Takahashi M, Tenen DG, Meyerson M, Wong KK, Shapiro GI (2008) Hsp90 inhibition suppresses mutant EGFR-T790 M signaling and overcomes kinase inhibitor resistance. Cancer Res 68(14):5827–5838. doi:10.1158/0008-5472.CAN-07-5428 CrossRefPubMedPubMedCentral

27.

Gong Y, Somwar R, Politi K, Balak M, Chmielecki J, Jiang X, Pao W (2007) Induction of BIM is essential for apoptosis triggered by EGFR kinase inhibitors in mutant EGFR-dependent lung adenocarcinomas. PLoS Med 4(10):e294. doi:10.1371/journal.pmed.0040294 CrossRefPubMedPubMedCentral

28.

Tjin Tham Sjin R, Lee K, Walter AO, Dubrovskiy A, Sheets M, Martin TS, Labenski MT, Zhu Z, Tester R, Karp R, Medikonda A, Chaturvedi P, Ren Y, Haringsma H, Etter J, Raponi M, Simmons AD, Harding TC, Niu D, Nacht M, Westlin WF, Petter RC, Allen A, Singh J (2014) In vitro and in vivo characterization of irreversible mutant-selective EGFR inhibitors that are wild-type sparing. Mol Cancer Ther 13(6):1468–1479. doi:10.1158/1535-7163.MCT-13-0966 CrossRefPubMed

29.

Ma C, Wei S, Song Y (2011) T790 M and acquired resistance of EGFR TKI: a literature review of clinical reports. Journal of thoracic disease 3(1):10–18. doi:10.3978/j.issn.2072-1439.2010.12.02 PubMedPubMedCentral

30.

Steuer CE, Khuri FR, Ramalingam SS (2014) The next generation of epidermal growth factor receptor tyrosine kinase inhibitors in the treatment of lung cancer. Cancer. doi:10.1002/cncr.29139

31.

Jorge SE, Kobayashi SS, Costa DB (2014) Epidermal growth factor receptor (EGFR) mutations in lung cancer: preclinical and clinical data. Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas /Sociedade Brasileira de Biofisica [et al] 47(11):929–939

32.

Steuer CE, Khuri FR, Ramalingam SS (2015) The next generation of epidermal growth factor receptor tyrosine kinase inhibitors in the treatment of lung cancer. Cancer 121(8):E1–E6. doi:10.1002/cncr.29139 CrossRefPubMed

33.

Sigismund S, Algisi V, Nappo G, Conte A, Pascolutti R, Cuomo A, Bonaldi T, Argenzio E, Verhoef LG, Maspero E, Bianchi F, Capuani F, Ciliberto A, Polo S, Di Fiore PP (2013) Threshold-controlled ubiquitination of the EGFR directs receptor fate. The EMBO journal 32(15):2140–2157. doi:10.1038/emboj.2013.149 CrossRefPubMedPubMedCentral

34.

Yang S, Qu S, Perez-Tores M, Sawai A, Rosen N, Solit DB, Arteaga CL (2006) Association with HSP90 inhibits Cbl-mediated down-regulation of mutant epidermal growth factor receptors. Cancer Res 66(14):6990–6997. doi:10.1158/0008-5472.CAN-06-1042 CrossRefPubMed

35.

Eck MJ, Yun CH (2010) Structural and mechanistic underpinnings of the differential drug sensitivity of EGFR mutations in non-small cell lung cancer. Biochim Biophys Acta 1804(3):559–566. doi:10.1016/j.bbapap.2009.12.010 CrossRefPubMed

36.

Kumar A, Petri ET, Halmos B, Boggon TJ (2008) Structure and clinical relevance of the epidermal growth factor receptor in human cancer. J Clin Oncol Off J Am Soc Clin Oncol 26(10):1742–1751. doi:10.1200/JCO.2007.12.1178 CrossRef

37.

Cho J, Chen L, Sangji N, Okabe T, Yonesaka K, Francis JM, Flavin RJ, Johnson W, Kwon J, Yu S, Greulich H, Johnson BE, Eck MJ, Janne PA, Wong KK, Meyerson M (2013) Cetuximab response of lung cancer-derived EGF receptor mutants is associated with asymmetric dimerization. Cancer Res 73(22):6770–6779. doi:10.1158/0008-5472.CAN-13-1145 CrossRefPubMedPubMedCentral

38.

Vacchelli E, Aranda F, Eggermont A, Galon J, Sautes-Fridman C, Zitvogel L, Kroemer G, Galluzzi L (2014) Trial Watch: Tumor-targeting monoclonal antibodies in cancer therapy. Oncoimmunology 3(1):e27048. doi:10.4161/onci.27048 CrossRefPubMedPubMedCentral

39.

Ahsan A, Ramanand SG, Bergin IL, Zhao L, Whitehead CE, Rehemtulla A, Ray D, Pratt WB, Lawrence TS, Nyati MK (2014) Efficacy of an EGFR-specific peptide against EGFR-dependent cancer cell lines and tumor xenografts. Neoplasia 16(2):105–114. doi:10.1593/neo.14182 CrossRefPubMedPubMedCentral

40.

Meador CB, Jin H, de Stanchina E, Nebhan CA, Pirazzoli V, Wang L, Lu P, Vuong H, Hutchinson KE, Jia P, Chen X, Eisenberg R, Ladanyi M, Politi K, Zhao Z, Lovly CM, Cross DA, Pao W (2014) Optimizing the Sequence of Anti-EGFR-Targeted therapy in EGFR-Mutant lung cancer. Mol Cancer Ther. doi:10.1158/1535-7163.MCT-14-0723 PubMedPubMedCentral

Title: AZD9291 overcomes T790 M-mediated resistance through degradation of EGFRL858R/T790M in non-small cell lung cancer cells
Authors: Bo Mi Ku
Yeon-Hee Bae
Jiae Koh
Jong-Mu Sun
Se-hoon Lee
Jin Seok Ahn
Keunchil Park
Myung-Ju Ahn
Publication date: 01-08-2016
Publisher: Springer US
Published in: Investigational New Drugs / Issue 4/2016
Print ISSN: 0167-6997
Electronic ISSN: 1573-0646
DOI: https://doi.org/10.1007/s10637-016-0350-y

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