Abstract 1591: Mechanisms of BPM31510-induced cell death in cancer: lipid peroxidation and apoptosis pathways

Abstract BPM31510, an oxidized Coenzyme Q10 (CoQ10) nanodispersion, has shown potential as an anticancer agent through its ability to induce the generation of reactive oxygen species (ROS) within the mitochondria of cancer cells. Recent studies have highlighted ferroptosis — a form of regulated cell death driven by iron-mediated lipid peroxidation, as a promising mechanism for cancer therapy, particularly for cancers that are highly vulnerable to oxidative stress. Ferroptosis differs from apoptosis in that it involves the accumulation of ROS, leading to lipid peroxidation and cell membrane damage, rather than DNA fragmentation and caspase activation characteristic of apoptosis. Given the strong correlation between BPM31510-induced cytotoxicity and elevated ROS levels, we investigated if BPM31510 induces cancer cell death through ferroptosis. To evaluate lipid peroxidation, we assessed both cancerous (HT1080, U87, GL261 and C6) and non-cancerous cells (NIH 3T3, TNC1, rat astrocytes, and human astrocytes). Lipid peroxidation was quantified using the fluorescent probe BODIPY 581/591. Cells were treated with BPM31510 and Erastin (a ferroptosis inducer) for 3 hours, and fluorescence intensity was measured to assess lipid peroxidation levels. To address whether BPM31510-induced cytotoxicity involves ferroptosis, we performed a cytotoxicity assay with additional conditions including Ferrostatin-1 (a ferroptosis inhibitor) combined with BPM31510 or Erastin. Inhibition of lipid peroxidation by Ferrostatin-1 would suggest ferroptosis involvement. Lastly, we assessed cell death pathways using annexin V and propidium iodide (PI) staining to distinguish apoptotic versus necrotic cell death in cells treated with BPM31510. BPM31510 treatment resulted in significantly elevated lipid peroxidation in cancer cell lines (HT1080, U87, Gl261, C6) compared to healthy cells (NIH 3T3, TNC1, astrocytes), suggesting differential vulnerability of cancer cells to BPM31510-induced oxidative stress. Although Ferrostatin-1 significantly reduced lipid peroxidation induced by Erastin, it did not reverse the lipid peroxidation caused by BPM31510. This resistance to Ferrostatin-1 indicates that BPM31510’s cytotoxic effects are independent of ferroptosis, distinguishing its mechanism from that of traditional ferroptosis inducers. Apoptosis assays further confirmed that BPM31510 predominantly activates apoptotic pathways, with limited necrotic cell death observed. Our findings demonstrate that BPM31510 differentially induces lipid peroxidation in cancer cells through a mechanism independent of ferroptosis, instead activating apoptosis as the primary pathway of cell death. The ferroptosis-independent cytotoxicity of BPM31510, combined with its selective targeting of cancer cells, highlights its potential as a unique therapeutic agent in glioblastoma. Citation Format: Abbas Khojasteh, Michael A. Kiebish, Lisa Ghiglieri, Sylwia Stopka, Juan J. Aristizabal-Henao, Seema Nagpal, Maria-D Nastke, Stephane Gesta, Niven R. Narain, Vijay Modur, Lawrence Recht. Mechanisms of BPM31510-induced cell death in cancer: lipid peroxidation and apoptosis pathways [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 1591.