RRC ID 44564
Author Byron SA, Chen H, Wortmann A, Loch D, Gartside MG, Dehkhoda F, Blais SP, Neubert TA, Mohammadi M, Pollock PM.
Title The N550K/H mutations in FGFR2 confer differential resistance to PD173074, dovitinib, and ponatinib ATP-competitive inhibitors.
Journal Neoplasia
Abstract We sought to identify fibroblast growth factor receptor 2 (FGFR2) kinase domain mutations that confer resistance to the pan-FGFR inhibitor, dovitinib, and explore the mechanism of action of the drug-resistant mutations. We cultured BaF3 cells overexpressing FGFR2 in high concentrations of dovitinib and identified 14 dovitinib-resistant mutations, including the N550K mutation observed in 25% of FGFR2(mutant) endometrial cancers (ECs). Structural and biochemical in vitro kinase analyses, together with BaF3 proliferation assays, showed that the resistance mutations elevate the intrinsic kinase activity of FGFR2. BaF3 lines were used to assess the ability of each mutation to confer cross-resistance to PD173074 and ponatinib. Unlike PD173074, ponatinib effectively inhibited all the dovitinib-resistant FGFR2 mutants except the V565I gatekeeper mutation, suggesting ponatinib but not dovitinib targets the active conformation of FGFR2 kinase. EC cell lines expressing wild-type FGFR2 were relatively resistant to all inhibitors, whereas EC cell lines expressing mutated FGFR2 showed differential sensitivity. Within the FGFR2(mutant) cell lines, three of seven showed marked resistance to PD173074 and relative resistance to dovitinib and ponatinib. This suggests that alternative mechanisms distinct from kinase domain mutations are responsible for intrinsic resistance in these three EC lines. Finally, overexpression of FGFR2(N550K) in JHUEM-2 cells (FGFR2(C383R)) conferred resistance (about five-fold) to PD173074, providing independent data that FGFR2(N550K) can be associated with drug resistance. Biochemical in vitro kinase analyses also show that ponatinib is more effective than dovitinib at inhibiting FGFR2(N550K). We propose that tumors harboring mutationally activated FGFRs should be treated with FGFR inhibitors that specifically bind the active kinase.
Volume 15(8)
Pages 975-88
Published 2013-8
DOI 10.1593/neo.121106
PMID 23908597
PMC PMC3730048
MeSH Adenosine Triphosphate / metabolism Animals Benzimidazoles / pharmacology* Binding Sites / genetics Biocatalysis / drug effects Cell Line Cell Line, Tumor Cell Proliferation / drug effects Drug Resistance, Neoplasm / drug effects Drug Resistance, Neoplasm / genetics Electrophoresis, Polyacrylamide Gel Endometrial Neoplasms / genetics Endometrial Neoplasms / metabolism Endometrial Neoplasms / pathology Female Humans Imidazoles / pharmacology* Inhibitory Concentration 50 Models, Molecular Mutation* Phosphorylation / drug effects Protein Kinase Inhibitors / pharmacology Protein Structure, Tertiary Pyridazines / pharmacology* Pyrimidines / pharmacology* Quinolones / pharmacology* Receptor, Fibroblast Growth Factor, Type 2 / chemistry Receptor, Fibroblast Growth Factor, Type 2 / genetics* Receptor, Fibroblast Growth Factor, Type 2 / metabolism
IF 3.837
Times Cited 35
Human and Animal Cells