Clofoctol as a novel senolytic drug eliminating therapy-induced senescent glioma cells.

Background: Glioblastoma is the most common malignant brain glioma, accounting for ∼48% of malignant central nervous system tumors. Targeting glioma stem cells and senescent glioma cells represents promising therapeutic strategies. In our previous study, we identified the clofoctol as a candidate drug targeting glioma stem cells with good blood-brain barrier permeability and potent anti-glioblastoma efficacy. Comprehensively demonstrating the impact of clofoctol on glioblastoma might provide novel strategies for the treatment of glioblastoma. Methods: By utilizing single-cell RNA sequencing of tumor tissue, we demonstrated the suppression effect of clofoctol on senescent glioma cell. Cellular RNA sequencing, molecular docking and CETSA were used to further confirm target of clofoctol. Ultimately, GL261 and orthotopic patient-derived xenograft animal models was performed to assess whether the senolytic effect of clofoctol could enhance TMZ therapy. Results: Clofoctol treatment reduced the senescence level (SASPs, senescence-related genes, and the proportion of senescent cells) in GL261-derived tumor single-cell RNA sequencing. In vitro, clofoctol could target senescent glioma cells and induce cell death through apoptosis and ferroptosis. P53 was identified as the functional protein which elicited the effect of clofoctol. In vivo, clofoctol exhibited senolytic activity and synergized with TMZ, leading to extended survival in glioblastoma mouse model. Conclusions: Our study demonstrated the clinical drug clofoctol could target chemotherapy-induced senescent glioma cells through P53 and trigger cell apoptotic and ferroptotic death. We further confirmed a synergistic effect between clofoctol and temozolomide which could be a novel therapeutic approach for glioblastoma therapy.