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Targeted Oncology

, Volume 14, Issue 5, pp 619–626 | Cite as

Acquired BRAF G469A Mutation as a Resistance Mechanism to First-Line Osimertinib Treatment in NSCLC Cell Lines Harboring an EGFR Exon 19 Deletion

  • Silvia La Monica
  • Roberta Minari
  • Daniele Cretella
  • Mara Bonelli
  • Claudia Fumarola
  • Andrea Cavazzoni
  • Maricla Galetti
  • Graziana Digiacomo
  • Federica Riccardi
  • Pier Giorgio Petronini
  • Marcello TiseoEmail author
  • Roberta Alfieri
Short Communication

Abstract

Background

Osimertinib is a new third-generation, epidermal growth factor receptor-tyrosine kinase inhibitor highly selective for the epidermal growth factor receptor with both activating and T790M mutations. A recent phase III trial showed a statistically significant progression-free survival benefit with osimertinib vs. gefitinib or erlotinib as first-line treatment for EGFR-mutated non-small cell lung cancer, and preliminary data are available on resistance mechanisms to first-line osimertinib therapy.

Objective

The objective of this study was to examine potential in vitro mechanisms of acquired resistance to osimertinib in a cell model carrying an EGFR exon 19 deletion.

Methods

PC9 cells were cultured in the presence of increasing concentrations of osimertinib (ranging from 10 to 500 nM) to generate resistant cells. Three clones resistant to osimertinib (half maximal inhibitory concentration > 1 μM) were isolated, genotyped by next-generation sequencing and tested for drug sensitivity. Cell proliferation and migration, cell death, and signaling transduction pathways were analyzed.

Results

Our study revealed that all the three resistant clones developed acquired resistance via the BRAF G469A mutation maintaining a constitutive activation of the ERK pathway. Stable transfection of PC9 and HCC827 cells with a plasmid containing BRAF G469A rendered the cells resistant to osimertinib. Treatment with selumetinib and trametinib, but not dabrafenib, restored the sensitivity to osimertinib and enhanced cell death in the resistant clones with the BRAF G469A mutation.

Conclusions

Our in vitro studies revealed the BRAF G469A-activating mutation as a potential mechanism of acquired resistance to first-line osimertinib treatment, and provide a strategy of intervention to overcome this mechanism of resistance.

Notes

Compliance with Ethical Standards

Funding

This work was supported by Associazione Italiana per la Ricerca sul Cancro (AIRC), Milan Grant IG2017-20074 (P.I. Marcello Tiseo). AstraZeneca provided a research grant to Marcello Tiseo.

Conflict of Interest

Marcello Tiseo has been on advisory boards and received speakers’ fees for Astra-Zeneca, Pfizer, Eli-Lilly, BMS, Novartis, Roche, MSD, Boehringer Ingelheim, Otsuka, Takeda, and Pierre Fabre. Marcello Tiseo has received research grants from Astra-Zeneca and Boehringer Ingelheim. Silvia La Monica, Roberta Minari, Daniele Cretella, Mara Bonelli, Claudia Fumarola, Andrea Cavazzoni, Maricla Galetti, Graziana Digiacomo, Federica Riccardi, Pier Giorgio Petronini, and Roberta Alfieri have no conflicts of interest that are directly relevant to the content of this article.

Supplementary material

11523_2019_669_MOESM1_ESM.pdf (120 kb)
Supplementary material 1 (PDF 120 kb)

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Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  • Silvia La Monica
    • 1
  • Roberta Minari
    • 2
  • Daniele Cretella
    • 1
  • Mara Bonelli
    • 1
  • Claudia Fumarola
    • 1
  • Andrea Cavazzoni
    • 1
  • Maricla Galetti
    • 3
    • 4
  • Graziana Digiacomo
    • 1
  • Federica Riccardi
    • 5
  • Pier Giorgio Petronini
    • 1
  • Marcello Tiseo
    • 1
    • 2
    Email author
  • Roberta Alfieri
    • 1
  1. 1.Department of Medicine and SurgeryUniversity of ParmaParmaItaly
  2. 2.Medical Oncology UnitUniversity Hospital of ParmaParmaItaly
  3. 3.Italian Workers’ Compensation Authority (INAIL) Research CenterParmaItaly
  4. 4.Center of Excellence for Toxicological Research (CERT)University of ParmaParmaItaly
  5. 5.Regional Reference Centre for Inherited Bleeding DisordersUniversity Hospital of ParmaParmaItaly

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