Abstract P2-02-15: All-trans retinoic acid perturbs the lipidomic profiles of luminal breast cancer cells characterized by sensitivity to the anti-proliferative activity of the retinoid

Abstract

Abstract Background: All-trans retinoic acid (ATRA) is the active metabolite of vitamin A and a promising agent in the prevention and treatment of breast cancer. We recently demonstrated that approximately 70% of estrogen-receptor-positive (ER+) breast cancer cell lines and primary tumors are sensitive to the anti-proliferative effects of ATRA (1,2). In contrast only 10-20% of the HER2-positive and triple-negative counterparts respond to the retinoid. The significance of lipids in the growth, progression and drug sensitivity of specific types of solid tumors, including breast cancer, is largely overlooked. In particular the role, if any, of specific lipids in the anti-tumor action of ATRA in breast cancer has never been studied. Aims and Approach: The principal aim of the present work was to evaluate the specific perturbations induced by ATRA on the homeostasis of lipids in breast cancer cells characterized by sensitivity to the anti-proliferative action of the retinoid. To this purpose, we took a high-throughput approach and defined the lipidomic profiles of 16 breast cancer cell lines in basal conditions and following challenge with ATRA (1 μM) for 48 hours. The panel consisted of eight cell lines characterized by a luminal phenotype and eight cell lines with a basal phenotype. Four ATRA-sensitive cell lines (SKBR3, HCC1500, CAMA1 and MDAMB361) and 4 ATRA-resistant counterparts (HCC202, MDAMB175VII, ZR75.1 and HCC1419) were included in the luminal group. Similarly, the basal group contained 4 ATRA-sensitive (HCC1599, MB157, MDAMB157 and Hs578T) and 4 ATRA-resistant (MDAMB231, CAL851, HCC1187 and MDAMB436) cell types. Results: Using Lipostar, a unique and recently developed software for high-throughput LC-MS lipidomics analysis (3), we identified lipid species whose levels were modified by ATRA in each cell line. This resulted in the generation of a lipid fingerprint consisting of 530 elements. We observed that ATRA reduced the amounts of cardiolipins in luminal and ATRA-sensitive breast cancer cell lines specifically. Similar effects were not observed in luminal and ATRA-resistant cells. ATRA-dependent reduction in the amounts of cardiolipins were never observed in basal cells, regardless of their sensitivity to the retinoid. Given the role played by cardiolipins in the homeostasis of the mitochondria, we evaluated the action of ATRA on the functional activity of these organelles in the luminal and ATRA-sensitive or ATRA-resistant cell lines. In SKBR3 and the other sensitive cell lines, we observed that ATRA modulated mitochondria-dependent oxygen consumption and ATP production. These effects were accompanied by an increase in mitochondrial membrane fluidity, which is consistent with the effects exerted by ATRA on the cellular content of cardiolipins. ATRA-dependent action on cardiolipins and mitochondrial homeostasis precedes maximal growth inhibition. The results provide new insights on the mechanisms underlying the anti-tumor action of ATRA. References : 1) Centritto F. et al. EMBO Mol Med. 2015 Jul;7(7):950-72. 2) Bolis M et al.. Ann Oncol. 2017 Mar 1;28(3):611-621. 3) Goracci, L. et al., Anal Chem 2017, 89 (11), 6258-6265. Citation Format: Terao M, Celestini V, Kurosaki M, Vallerga A, Bolis M, Fratelli M, Paroni G, Di Veroli A, Cruciani G, Goracci L, Garattini E. All-trans retinoic acid perturbs the lipidomic profiles of luminal breast cancer cells characterized by sensitivity to the anti-proliferative activity of the retinoid [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-02-15.