Dual ALK and EGFR inhibition targets a mechanism of acquired resistance to the tyrosine kinase inhibitor crizotinib in ALK rearranged lung cancer

Norihiro Yamaguchi, Antonio R. Lucena-Araujo, Sohei Nakayama, Lorena L. de Figueiredo-Pontes, David A. Gonzalez, Hiroyuki Yasuda, Susumu Kobayashi, Daniel B. Costa

Research output: Contribution to journalArticle

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Abstract

Introduction: The multitargeted tyrosine kinase inhibitor (TKI) crizotinib is active against ALK translocated non-small-cell lung cancer (NSCLC); however acquired resistance invariably develops over time. ALK mutations have previously been implicated in only a third of resistant tumors. We sought to evaluate alternative mechanisms of resistance and preclinical strategies to overcome these in a cell line driven by EML4-ALK. Methods: We selected the NSCLC cell line NCI-H3122 (H3122: EML4- ALK E13;A20) and derived resistant variants that were able to grow in the presence of 1. μM crizotinib. These were analyzed for ALK mutations, sensitivity to crizotinib in combination with other TKIs, and for activation of alternative tyrosine kinases. Results: All H3122 crizotinib resistant (CR) clones lacked amplification or mutations in the kinase domain of ALK. To evaluate if possible alternative kinases functioned as "bypass" tracks for downstream signaling activation in these resistance cells, we performed of phosho-receptor tyrosine kinase array that demonstrated that CR clones had higher phospho-EGFR signals than H3122 cells before and after exposure to crizotinib. A functional approach of dual ALK TKI (with crizotinib) with combinatory TKI inhibition was used as a secondary screen for possible targets. Crizotinib. +. erlotinib (reversible EGFR TKI) and crizotinib. +. afatinib (irreversible EGFR/ERBB2 TKI) were able to inhibit the growth of H3122 CR clones, confirming EGFR activation as a mechanism of resistance. The removal of crizotinib from the culture media re-sensitized CR cells to crizotinib. Conclusions: We identified activation of EGFR as a mechanism of resistance to crizotinib in preclinical models of ALK translocated NSCLC. If EGFR activation is confirmed as a predominant mechanism of ALK TKI-induced resistance in patient-derived tumors, the use of ALK plus EGFR TKIs could be explored for this important cohort of NSCLCs.

Original languageEnglish
Pages (from-to)37-43
Number of pages7
JournalLung Cancer
Volume83
Issue number1
DOIs
Publication statusPublished - 2014 Jan
Externally publishedYes

Fingerprint

Protein-Tyrosine Kinases
Lung Neoplasms
Non-Small Cell Lung Carcinoma
Clone Cells
crizotinib
Mutation
Phosphotransferases
Cell Line
Receptor Protein-Tyrosine Kinases
Culture Media
Neoplasms

Keywords

  • Anaplastic lymphoma kinase
  • Crizotinib
  • Epidermal growth factor receptor
  • Kinase inhibitor
  • Non-small-cell lung cancer
  • Tyrosine kinase

ASJC Scopus subject areas

  • Oncology
  • Pulmonary and Respiratory Medicine
  • Cancer Research

Cite this

Dual ALK and EGFR inhibition targets a mechanism of acquired resistance to the tyrosine kinase inhibitor crizotinib in ALK rearranged lung cancer. / Yamaguchi, Norihiro; Lucena-Araujo, Antonio R.; Nakayama, Sohei; de Figueiredo-Pontes, Lorena L.; Gonzalez, David A.; Yasuda, Hiroyuki; Kobayashi, Susumu; Costa, Daniel B.

In: Lung Cancer, Vol. 83, No. 1, 01.2014, p. 37-43.

Research output: Contribution to journalArticle

Yamaguchi, N, Lucena-Araujo, AR, Nakayama, S, de Figueiredo-Pontes, LL, Gonzalez, DA, Yasuda, H, Kobayashi, S & Costa, DB 2014, 'Dual ALK and EGFR inhibition targets a mechanism of acquired resistance to the tyrosine kinase inhibitor crizotinib in ALK rearranged lung cancer', Lung Cancer, vol. 83, no. 1, pp. 37-43. https://doi.org/10.1016/j.lungcan.2013.09.019
Yamaguchi, Norihiro ; Lucena-Araujo, Antonio R. ; Nakayama, Sohei ; de Figueiredo-Pontes, Lorena L. ; Gonzalez, David A. ; Yasuda, Hiroyuki ; Kobayashi, Susumu ; Costa, Daniel B. / Dual ALK and EGFR inhibition targets a mechanism of acquired resistance to the tyrosine kinase inhibitor crizotinib in ALK rearranged lung cancer. In: Lung Cancer. 2014 ; Vol. 83, No. 1. pp. 37-43.
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T1 - Dual ALK and EGFR inhibition targets a mechanism of acquired resistance to the tyrosine kinase inhibitor crizotinib in ALK rearranged lung cancer

AU - Yamaguchi, Norihiro

AU - Lucena-Araujo, Antonio R.

AU - Nakayama, Sohei

AU - de Figueiredo-Pontes, Lorena L.

AU - Gonzalez, David A.

AU - Yasuda, Hiroyuki

AU - Kobayashi, Susumu

AU - Costa, Daniel B.

PY - 2014/1

Y1 - 2014/1

N2 - Introduction: The multitargeted tyrosine kinase inhibitor (TKI) crizotinib is active against ALK translocated non-small-cell lung cancer (NSCLC); however acquired resistance invariably develops over time. ALK mutations have previously been implicated in only a third of resistant tumors. We sought to evaluate alternative mechanisms of resistance and preclinical strategies to overcome these in a cell line driven by EML4-ALK. Methods: We selected the NSCLC cell line NCI-H3122 (H3122: EML4- ALK E13;A20) and derived resistant variants that were able to grow in the presence of 1. μM crizotinib. These were analyzed for ALK mutations, sensitivity to crizotinib in combination with other TKIs, and for activation of alternative tyrosine kinases. Results: All H3122 crizotinib resistant (CR) clones lacked amplification or mutations in the kinase domain of ALK. To evaluate if possible alternative kinases functioned as "bypass" tracks for downstream signaling activation in these resistance cells, we performed of phosho-receptor tyrosine kinase array that demonstrated that CR clones had higher phospho-EGFR signals than H3122 cells before and after exposure to crizotinib. A functional approach of dual ALK TKI (with crizotinib) with combinatory TKI inhibition was used as a secondary screen for possible targets. Crizotinib. +. erlotinib (reversible EGFR TKI) and crizotinib. +. afatinib (irreversible EGFR/ERBB2 TKI) were able to inhibit the growth of H3122 CR clones, confirming EGFR activation as a mechanism of resistance. The removal of crizotinib from the culture media re-sensitized CR cells to crizotinib. Conclusions: We identified activation of EGFR as a mechanism of resistance to crizotinib in preclinical models of ALK translocated NSCLC. If EGFR activation is confirmed as a predominant mechanism of ALK TKI-induced resistance in patient-derived tumors, the use of ALK plus EGFR TKIs could be explored for this important cohort of NSCLCs.

AB - Introduction: The multitargeted tyrosine kinase inhibitor (TKI) crizotinib is active against ALK translocated non-small-cell lung cancer (NSCLC); however acquired resistance invariably develops over time. ALK mutations have previously been implicated in only a third of resistant tumors. We sought to evaluate alternative mechanisms of resistance and preclinical strategies to overcome these in a cell line driven by EML4-ALK. Methods: We selected the NSCLC cell line NCI-H3122 (H3122: EML4- ALK E13;A20) and derived resistant variants that were able to grow in the presence of 1. μM crizotinib. These were analyzed for ALK mutations, sensitivity to crizotinib in combination with other TKIs, and for activation of alternative tyrosine kinases. Results: All H3122 crizotinib resistant (CR) clones lacked amplification or mutations in the kinase domain of ALK. To evaluate if possible alternative kinases functioned as "bypass" tracks for downstream signaling activation in these resistance cells, we performed of phosho-receptor tyrosine kinase array that demonstrated that CR clones had higher phospho-EGFR signals than H3122 cells before and after exposure to crizotinib. A functional approach of dual ALK TKI (with crizotinib) with combinatory TKI inhibition was used as a secondary screen for possible targets. Crizotinib. +. erlotinib (reversible EGFR TKI) and crizotinib. +. afatinib (irreversible EGFR/ERBB2 TKI) were able to inhibit the growth of H3122 CR clones, confirming EGFR activation as a mechanism of resistance. The removal of crizotinib from the culture media re-sensitized CR cells to crizotinib. Conclusions: We identified activation of EGFR as a mechanism of resistance to crizotinib in preclinical models of ALK translocated NSCLC. If EGFR activation is confirmed as a predominant mechanism of ALK TKI-induced resistance in patient-derived tumors, the use of ALK plus EGFR TKIs could be explored for this important cohort of NSCLCs.

KW - Anaplastic lymphoma kinase

KW - Crizotinib

KW - Epidermal growth factor receptor

KW - Kinase inhibitor

KW - Non-small-cell lung cancer

KW - Tyrosine kinase

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