Early Ferroptosis Detection Targeting Lipid Hydrogen Abstraction

Ferroptosis is an Fe2+-dependent and lipid peroxidation-mediated regulated cell death. Its early detection is critical for diagnosis and timely intervention. Lipid hydrogen abstraction is the initiation step of the "lipid peroxidation-ferroptosis" cascade and an ideal target for early detection of ferroptosis. We developed the first set of fluorogenic probes for lipid H-abstraction, i.e., LHA585 for red, LHA675 for deep-red, and LHA930 for near-infrared. The key to their design is the use of 4-phenyl-3-methylbut-2-enyl, a close mimic of the polyunsaturated lipid, to specifically detect the highly oxidative radicals involved in ferroptosis. Notably, LHA585 yielded a fluorescence turn-on ca. 8 h earlier than the current gold-standard probe in an in vitro OGD/R model, highlighting its superiority for early detection of ferroptosis. While LHA585/675 is intended for in vitro studies, LHA930 was feasible for in vivo ferroptosis detection. Collectively, these findings establish LHAs as a robust advance for ferroptosis sensing, enabling in-depth mechanistic studies of lipid-peroxidation-driven cell death.