Abstract A117: Evaluation of nab-sirolimus in combination with PI3K pathway inhibitors to overcome PI3K/mTOR resistance in PI3K-mutant breast cancer cell lines

Abstract

Abstract Background: The PI3K-AKT-mTOR pathway is frequently activated in many cancer types and plays a central role in tumorigenesis. However, clinical use of therapies targeting this pathway is limited by compensatory vertical and horizontal feedback activation loops, which limit antitumor efficacy and lead to drug resistance. Combination therapy strategies employing simultaneous inhibition of multiple PI3K-AKT-mTOR pathway elements may overcome these resistance mechanisms and improve antitumor activity. nab-Sirolimus is a novel albumin-bound nanoparticle form of the mTOR inhibitor sirolimus that has significantly greater tumor accumulation, mTOR target suppression, and improved antitumor effects compared with conventional first generation mTOR inhibitors in preclinical models. Here, we analyzed the effects of nab-sirolimus combinations in PI3K-mutant breast cancer (BrCa) cells. Methods: PI3K mutated BrCa cell lines (MDA-MD-453, hormone receptor [HR]-; and MDA-MB-361, HR+) were incubated for 5 days with increasing concentrations of nab-sirolimus, the PI3K pathway inhibitors gedatolisib and alpelisib, or the AKT inhibitors miransertinib and capivasertib. The antiproliferative and cytotoxic effects of single agent and combination treatment were assessed using cell viability and cell death assays. Cell signaling was analyzed by western blot. Results: The antiproliferative effects of gedatolisib at low doses (2.5-10 nM) were enhanced by 61-71% in combination with nab-sirolimus (20 or 80 nM) in MDA-MB-453 cells. Limited cell death (4.2-9.6%) was observed in HR- MDA-MB-453 cells with low doses (2.5, 5, and 10 nM) of gedatolisib alone. In contrast, and despite the well-established cytostatic action of mTOR inhibitors, the addition of 20 and 80 nM nab-sirolimus led to high levels of cell death (up to 48.9%). Similar results were observed with alpelisib at 1 µM in combination with nab-sirolimus (20 or 80 nM), and also with capivasertib (1 µM) or miransertib (250 nM) combined with 80 nM nab-sirolimus. Western blot analysis demonstrated that gedatolisib or alpelisib alone each triggered upregulation of p4EBP1 whereas nab-sirolimus treatment alone activated a negative feedback loop through increased pAKT. Importantly, these compensatory feedback activation loops were both reversed by the combination of PI3K inhibitors and nab-sirolimus. Similar cell death and cell viability results were observed with nab-sirolimus plus PI3K inhibitors (gedatolisib or alpelisib) and AKT inhibitors (capivasertib or miransertib) in HR+ MDA-MB-361 cells. Conclusions: The antitumor effects of PI3K and AKT inhibitors were enhanced by the addition of the novel mTOR inhibitor nab-sirolimus. The improved effectiveness of the PI3K and mTOR inhibitor combination was due to the reciprocal overcoming of resistance mechanisms induced by single agent treatment. These data support a vertical PI3K pathway inhibition strategy using nab-sirolimus and PI3K/AKT inhibitors in PIK3CA-mutated BrCa, regardless of HR status. Citation Format: Sean Wallace, Khine Nyein Myint, Shihe Hou, Maria Zalath, Andrew Kwon, Brian McMorran, Igor Vivanco. Evaluation of nab-sirolimus in combination with PI3K pathway inhibitors to overcome PI3K/mTOR resistance in PI3K-mutant breast cancer cell lines [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr A117.