Acquired resistance to an epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) in an uncommon G719S EGFR mutation

  • Atsushi Osoegawa
  • Takafumi Hashimoto
  • Yohei Takumi
  • Miyuki Abe
  • Tomonori Yamada
  • Ryoji Kobayashi
  • Michiyo Miyawaki
  • Hideya Takeuchi
  • Tatsuro Okamoto
  • Kenji Sugio
PRECLINICAL STUDIES
  • 82 Downloads

Summary

Background Acquired resistance (AR) to an epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) is a common event, and several underlying mechanisms, including T790 M, MET amplification and PTEN downregulation, have been reported for the common EGFR mutations. EGFR G719X is an uncommon mutation that has been reported to show sensitivity to EGFR-TKIs. However, no established cell lines harboring the EGFR G719X have been reported in the literature. Materials and Methods G719S-GR cells were established from malignant pleural effusion of a patient whose tumor developed AR from gefitinib treatment. G719S-GR cells were then genotyped and tested for drug sensitivities. Multiplex ligation-dependent probe amplification (MLPA) was used to compare the clinical tumor samples with G719S-GR. Results G719S-GR cells were resistant to EGFR-TKIs with an LC50 of around 10 μM. A genomic analysis showed that G719S-GR cells harbor the EGFR G719S mutation as well as the amplification of EGFR locus. The homozygous deletion of CDKN2A and the loss of PTEN and TSC1 were also detected. On comparing the copy number of tumor suppressor genes using MLPA, G719S-GR cells were found to lack one copy of PTEN, which was not observed in a tumor obtained before gefitinib treatment. Loss of PTEN may result in AKT activation. The mTORC1/2 inhibitor Torin-1 was able to inhibit the downstream signaling when combined with osimertinib. Discussion The newly established G719S-GR cell line may be useful for investigating the mechanism underlying the development of AR in the G719X mutation; the loss of PTEN may be one such mechanism.

Keywords

EGFR Acquired resistance EGFR G719X Osimertinib Torin-1 

Abbreviations

TKI

tyrosine kinase inhibitor

EGFR

epidermal growth factor receptor

AR

acquired resistance

MPE

malignant pleural effusion

ROCK

Rho-associated coiled-coil forming kinase

MLPA

multiplex ligation-dependent probe amplification

GR

gefitinib resistant

LC50

Lethal Concentration, 50%

Notes

Acknowledgements

The authors would like to thank Mr. Brian Quinn for his critical comments on the manuscript.

Compliance with ethical standards

Conflict of interest

AO declares that he has no conflict of interest. TH declares that he has no conflict of interest. YT declares that he has no conflict of interest. MA declares that she has no conflict of interest. TY declares that he has no conflict of interest. RK declares that he has no conflict of interest. MM declares that she has no conflict of interest. HT declares that he has no conflict of interest. TO declares that he has no conflict of interest. KS declares that he has no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study.

Supplementary material

10637_2018_592_Fig4_ESM.gif (203 kb)
Supplemental Figure 1

Genomic alteration in G719S-GR cells. Copy number variations were estimated by comparing with NCC oncopanel data obtained from four normal tissues as a control. The amplification of EGFR, IL7R, MYC and the FGFR1 locus was observed. The homozygous deletion of CDKN2A and the loss of PTEN and TSC1 were also detected. (GIF 203 kb)

10637_2018_592_MOESM1_ESM.tif (2.7 mb)
High resolution image (TIFF 2765 kb)
10637_2018_592_MOESM2_ESM.docx (27 kb)
Supplemental Table 1 (DOCX 27 kb)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Atsushi Osoegawa
    • 1
  • Takafumi Hashimoto
    • 1
  • Yohei Takumi
    • 1
  • Miyuki Abe
    • 1
  • Tomonori Yamada
    • 1
  • Ryoji Kobayashi
    • 1
  • Michiyo Miyawaki
    • 1
  • Hideya Takeuchi
    • 1
  • Tatsuro Okamoto
    • 1
  • Kenji Sugio
    • 1
  1. 1.Department of Thoracic and Breast SurgeryOita University Faculty of MedicineYufuJapan

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