Targeting FGFR overcomes EMT-mediated resistance in EGFR mutant non-small cell lung cancer

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Abstract

Evolved resistance to tyrosine kinase inhibitor (TKI)-targeted therapies remains a major clinical challenge. In epidermal growth factor receptor (EGFR) mutant non-small-cell lung cancer (NSCLC), failure of EGFR TKIs can result from both genetic and epigenetic mechanisms of acquired drug resistance. Widespread reports of histologic and gene expression changes consistent with an epithelial-to-mesenchymal transition (EMT) have been associated with initially surviving drug-tolerant persister cells, which can seed bona fide genetic mechanisms of resistance to EGFR TKIs. While therapeutic approaches targeting fully resistant cells, such as those harboring an EGFRT790M mutation, have been developed, a clinical strategy for preventing the emergence of persister cells remains elusive. Using mesenchymal cell lines derived from biopsies of patients who progressed on EGFR TKI as surrogates for persister populations, we performed whole-genome CRISPR screening and identified fibroblast growth factor receptor 1 (FGFR1) as the top target promoting survival of mesenchymal EGFR mutant cancers. Although numerous previous reports of FGFR signaling contributing to EGFR TKI resistance in vitro exist, the data have not yet been sufficiently compelling to instigate a clinical trial testing this hypothesis, nor has the role of FGFR in promoting the survival of persister cells been elucidated. In this study, we find that combining EGFR and FGFR inhibitors inhibited the survival and expansion of EGFR mutant drug-tolerant cells over long time periods, preventing the development of fully resistant cancers in multiple vitro models and in vivo. These results suggest that dual EGFR and FGFR blockade may be a promising clinical strategy for both preventing and overcoming EMT-associated acquired drug resistance and provide motivation for the clinical study of combined EGFR and FGFR inhibition in EGFR-mutated NSCLCs.

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Acknowledgements

We thank Dr. Rafael Irizarry and Irineo Cabreros for guidance in statistical analyses related to shRNA dropout screening. We also thank Drs. Cyril Benes, Daniel Haber, Joan Brugge, Kornelia Polyak, and Robert Weinberg for helpful discussions throughout the development of this work. This study was funded by support from the NIH F30 CA213726-01A1 (SR), K08CA197389 (ANH), R01CA137008 (LVS), Doris Duke Charitable Foundation (ANH), Stand Up To Cancer (ANH and LVS), National Science Foundation (JAE), V Foundation (JAE), Ludwig Cancer Research (ANH), LungStrong, and Be a Piece of the Solution.

Author information

SR, IJM, MJN, JAE, and ANH designed the study, analyzed the data and wrote the paper. SR, IJM, HFC, VN, EL, NH, LD, JMK, SS and JB performed cell line and biochemical studies. SR, DT, and SJB performed tumor xenograft studies. LVS and ZP provided EGFR mutant patient samples. MG, AW, and KK, generated patient-derived cell lines. DAR, YD, and FJ performed computational analysis. All authors discussed the results and commented on the manuscript.

Correspondence to Aaron N. Hata.

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Conflict of interest

IJM, EL, NH, JMK, RT, DAR, PSH, MJN, JB, and JAE are employees of Novartis Inc., as noted in the affiliations. MJN and JAE hold equity interest in Novartis Inc. ANH receives research support from Novartis, Amgen, Pfizer, and Relay Therapeutics. ZP is a consultant/advisory board member for Takeda, AstraZeneca, GuardantHealth and Novartis, and receives institutional research support from Novartis. LVS is a consultant for AstraZeneca, Boehringer-Ingelheim, Novartis, Pfizer, Genentech, Merrimack, and BMS. The other authors declare that they have no conflict of interest.

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