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>i.e.</i>, <b>LHA</b>585 for red, <b>LHA675</b> for deep-red, and <b>LHA930</b> 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, <b>LHA585</b> yielded a fluorescence turn-on ca. 8 h earlier than the current gold-standard probe in an <i>in vitro</i> OGD/R model, highlighting its superiority for early detection of ferroptosis. While <b>LHA585/675</b> is intended for <i>in vitro</i> studies, <b>LHA930</b> was feasible for <i>in vivo</i> ferroptosis detection. Collectively, these findings establish <b>LHA</b>s as a robust advance for ferroptosis sensing, enabling in-depth mechanistic studies of lipid-peroxidation-driven cell death.