Construction of a novel cell-based assay for the evaluation of anti-EGFR drug efficacy against EGFR mutation

Hirotaka Hoshi, Gen Hiyama, Kosuke Ishikawa, Kiyoshi Inageda, Jiro Fujimoto, Ai Wakamatsu, Takushi Togashi, Yoshifumi Kawamura, Nobuhiko Takahashi, Arisa Higa, Naoki Goshima, Kentaro Semba, Shinya Watanabe, Motoki Takagi

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Epidermal growth factor receptor (EGFR) overexpression and EGFR-mediated signaling pathway dysregulation have been observed in tumors from patients with various cancers, especially non-small cell lung cancer. Thus, several anti-EGFR drugs have been developed for cancer therapy. For patients with known EGFR activating mutations (EGFR exon 19 in-frame deletions and exon 21 L858R substitution), treatment with an EGFR tyrosine kinase inhibitor (EGFR TKI; gefitinib, erlotinib or afatinib) represents standard first-line therapy. However, the clinical efficacy of these TKIs is ultimately limited by the development of acquired drug resistance such as by mutation of the gatekeeper T790 residue (T790M). To overcome this acquired drug resistance and develop novel anti-EGFR drugs, a cell-based assay system for EGFR TKI resistance mutant-selective inhibitors is required. We constructed a novel cell-based assay for the evaluation of EGFR TKI efficacy against EGFR mutation. To this end, we established non-tumorigenic immortalized breast epithelial cells that proliferate dependent on EGF (MCF 10A cells), which stably overexpress mutant EGFR. We found that the cells expressing EGFR containing the T790M mutation showed higher resistance against gefitinib, erlotinib and afatinib compared with cells expressing wild-type EGFR. In contrast, L858R mutant-expressing cells exhibited higher TKI sensitivity. The effect of T790M-selective inhibitors (osimertinib and rociletinib) on T790M mutant-expressing cells was significantly higher than gefitinib and erlotinib. Finally, when compared with commercially available isogenic MCF 10A cell lines carrying introduced mutations in EGFR, our EGFR mutant-overexpressing cells exhibited obviously higher responsiveness to EGFR TKIs depending on the underlying mutations because of the higher levels of EGFR phosphorylation in the EGFR mutant-overexpressing cells than in the isogenic cell lines. In conclusion, we successfully developed a novel cell-based assay for evaluating the efficacy of anti-EGFR drugs against EGFR mutation.

Original languageEnglish
Pages (from-to)66-76
Number of pages11
JournalOncology Reports
Volume37
Issue number1
DOIs
Publication statusPublished - 2017 Jan

Keywords

  • EGFR mutation
  • EGFR tyrosine kinase inhibitor
  • Epidermal growth factor receptor
  • Lung cancer
  • Molecular target drug
  • Non-small cell lung cancer
  • T790M

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Fingerprint Dive into the research topics of 'Construction of a novel cell-based assay for the evaluation of anti-EGFR drug efficacy against EGFR mutation'. Together they form a unique fingerprint.

  • Cite this

    Hoshi, H., Hiyama, G., Ishikawa, K., Inageda, K., Fujimoto, J., Wakamatsu, A., Togashi, T., Kawamura, Y., Takahashi, N., Higa, A., Goshima, N., Semba, K., Watanabe, S., & Takagi, M. (2017). Construction of a novel cell-based assay for the evaluation of anti-EGFR drug efficacy against EGFR mutation. Oncology Reports, 37(1), 66-76. https://doi.org/10.3892/or.2016.5227