CK-666 protects against ferroptosis and renal ischemia-reperfusion injury through a microfilament-independent mechanism

Abstract Ferroptosis is a type of regulated cell death caused by iron-dependent accumulation of lipid peroxidation, exhibiting unique morphological changes. Actin microfilaments are crucial for various cellular processes, including morphogenesis, motility, endocytosis, and cell death. However, the role of actin microfilaments in ferroptosis is not well understood. Here, we found that actin microfilaments undergo remodeling and disassembly during ferroptosis. Interestingly, inhibitors targeting actin microfilament remodeling did not impact cell sensitivity to ferroptosis, except for CK-666. Notably, CK-666 attenuated ferroptosis independently of its canonical function of inhibiting the ARP2/3 complex. Further investigation revealed that CK-666 modulates the ferroptosis proteome and prevents lipid degradation during ferroptosis. Liquid chromatography-mass spectrometry analysis and functional assays demonstrated that CK-666 mitigates ferroptosis by directly eliminating lipid peroxidation. Importantly, CK-666 significantly ameliorated renal ischemia-reperfusion injury and ferroptosis in renal tissue. In summary, our findings provide new insights into the relationship between cytoskeleton and ferroptosis, and suggest potential applications of CK-666 in the treatment of ferroptosis-related diseases.