CK-666 protects against ferroptosis and renal ischemia-reperfusion injury through a microfilament-independent mechanism
Ferroptosis is a form of regulated cell death characterized by iron-dependent lipid peroxidation and distinct morphological changes. While actin microfilaments play essential roles in processes like morphogenesis, motility, endocytosis, and cell death, their involvement in ferroptosis is not well understood. In this study, we demonstrate that actin microfilaments undergo remodeling and disassembly during ferroptosis. Surprisingly, inhibitors targeting actin microfilament remodeling do not influence the sensitivity of cells to ferroptosis, except for CK-666 and its structural analog CK-636. Mechanistically, CK-666 reduces ferroptosis without affecting its known role in inhibiting the Arp2/3 complex. Further analysis shows that CK-666 alters the ferroptotic transcriptome, prevents lipid degradation, and reduces lipid peroxidation. Additionally, CK-666 does not affect the labile iron pool within cells, and FSP1 inhibition does not alter its anti-ferroptotic effects. Notably, both DPPH and liposome leakage assays indicate that CK-666 directly reduces lipid peroxidation, mitigating ferroptosis. Importantly, CK-666 significantly improved renal ischemia-reperfusion injury and ferroptosis in renal tissue, highlighting its potential as a therapeutic agent.