Abstract P4-03-12: Effects of PI3K inhibitors on endocrine-resistant cell lines and differences in the characteristics of ER positive breast cancer cells after acquired resistance to the inhibitors

Abstract

Abstract Background Mutations in phosphatidylinositol 3-kinase (PI3K), which encodes the catalytic subunit of PI3Kα, is one of the most frequent genomic alterations and is found in about 40% of estrogen receptor (ER)-positive, HER2 negative breast cancers. PIK3CA mutations promote the growth and proliferation of cancers via activation of the PI3K/mammalian target of rapamycin pathway and can mediate resistance to endocrine therapies in breast cancer. Although several clinical trials for PI3K inhibitors (PI3Kis) in ER-positive metastatic breast cancers are ongoing, the effects of PI3Kis on endocrine-resistant breast cancers with various characteristics and definitive biomarkers of PI3Kis are unclear. Using endocrine-resistant cells established in our laboratory, we evaluated the efficacy of PI3Ki in these cell lines and identified the characteristics associated with acquired resistance to PI3Kis in endocrine-resistant cells. Results Long-term estrogen deprivation-resistant (EDR) cell lines and fulvestrant-resistant cell lines (MFR and TFR) were established from MCF-7 and T-47D cells in our previous studies. These cell lines showed different ER expression levels, including high expression (EDR-1), low expression (EDR-2, -3), or no expression (MFR, TFR); all of these cell lines had the same PIK3CA mutations as the parental cell lines. The pan-class1 PI3Ki buparlisib (BKM120) and α-specific PI3K inhibitor alpelisib (BYL719) inhibited the proliferation of endocrine-resistant cell lines when compared with their parental cells. Among endocrine-resistant cells, MFR cells were dramatically inhibited by PI3Kis. Colony formation assays indicated that MFR cells were more sensitive to PI3Kis than other cells lines. Next, we established PI3Ki- and everolimus-resistant cell lines from EDR-1, EDR-2, and MFR cells using BKM120, BYL719, and everolimus. Analysis of the time required to generate resistant cells showed that MFR cells required twice as long to acquire resistance compared with EDR cells. Furthermore, the time required to acquire resistance to BYL719 was shorter than that for BKM120. BYL719-resistant (BYL-R) cells were effectively inhibited by BKM120 to a degree similar to that of parental cells; however, BYL-R cells lost sensitivity to BYL719 and everolimus. Evelolimus-resistant (EVE-R) cells were also the same. In contrast, BKM120-resistant (BKM-R) cells showed less sensitivity to BKM120, BYL719, and everolimus. In other words, the pan-PI3Ki BKM120 was able to inhibit the growth of BYL-R and EVE-R cells, whereas BYL719 and everolimus were not able to inhibit BKM-R cells sufficiently. In addition, there were no changes in ER expression in EDR-1, EDR-2, and MFR cells exposed to PI3Kis for 1 h. Interestingly, ER expression on EDR-2 cells with acquired resistance to PI3Kis was increased compared with that in parental cells. Conclusion Our findings showed that PI3Kis exhibited remarkable efficacy in all types of EDR cells, particularly in fulvestrant-resistant cell lines. In PI3Ki- and everolimus-resistant cell lines, BKM120 and BYL719 showed different effects, and BYL719 and everolimus may exhibit cross-resistance. Furthermore, PI3Kis were likely to change the expression of ER. Citation Format: Tokuda E, Komatsu T, Sasaki S, Tsuboi K, Nakamura M, Iida M, Niwa T, Saito M, Hayashi S-I. Effects of PI3K inhibitors on endocrine-resistant cell lines and differences in the characteristics of ER positive breast cancer cells after acquired resistance to the inhibitors [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P4-03-12.