Ovarian carcinomas with mutations in the tumour suppressor BRCA2 are particularly sensitive to platinum compounds1. However, such carcinomas ultimately develop cisplatin resistance. The mechanism of that resistance is largely unknown2. Here we show that acquired resistance to cisplatin can be mediated by secondary intragenic mutations in BRCA2 that restore the wild-type BRCA2 reading frame. First, in a cisplatin-resistant BRCA2-mutated breast-cancer cell line, HCC1428, a secondary genetic change in BRCA2 rescued BRCA2 function. Second, cisplatin selection of a BRCA2-mutated pancreatic cancer cell line, Capan-1 (refs 3, 4), led to five different secondary mutations that restored the wild-type BRCA2 reading frame. All clones with secondary mutations were resistant both to cisplatin and to a poly(ADP-ribose) polymerase (PARP) inhibitor (AG14361). Finally, we evaluated recurrent cancers from patients whose primary BRCA2-mutated ovarian carcinomas were treated with cisplatin. The recurrent tumour that acquired cisplatin resistance had undergone reversion of its BRCA2 mutation. Our results suggest that secondary mutations that restore the wild-type BRCA2 reading frame may be a major clinical mediator of acquired resistance to platinum-based chemotherapy.
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We thank M.C. King and C. W. Drescher for discussions, B. Trask for overseeing the FISH analyses in her laboratory and for comments on the manuscript, J.W. Huang for comments on the manuscript, and M. Hoatlin and K. Polyak for reagents. We thank Pfizer for AG14361. We thank the Pacific Ovarian Cancer Research Consortium (supported by a Specialized Program of Research Excellence in Ovarian Cancer) for clinical specimens. This work was supported by grants from the National Institutes of Health/National Cancer Institute (to T.T. and E.M.S.), the Searle Scholars Program, the V Foundation and the Hartwell Innovation Fund (to T.T.), the L&S Milken Foundation (to B.Y.K.), the American Cancer Society California Division-Early Detection Professorship (to B.Y.K.), start-up funds from the Fred Hutchinson Cancer Research Center (to T.T.) and a gift from the Yvonne Betson Trust (to E.M.S.).
Author Contributions W.S. performed most of the experiments. E.M.S., B.Y.K. and N.U. provided clinical samples and expertise on ovarian cancer. E.M.S. performed laser capture microdissection and DNA extractions. M.K.A., D.J.F. and F.J.C. performed homologous recombination assays. J.H. sequenced BRCA2 in HCC1428. C.F. performed FISH analysis. E.V. performed the siRNA experiments shown in Fig. 1f, g. C.J. performed the PARP inhibitor sensitivity assays in Fig. 3c. T.T., W.S. and E.M.S. wrote the manuscript.
The file contains Supplementary Figures S1-S9 with Legends and Supplementary Tables S1-S5. (PDF 4082 kb)
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Angewandte Chemie International Edition (2019)
Molecular Cell (2019)
Genetics and Molecular Biology (2019)
Genes and Environment (2019)
Cell Cycle (2019)