Targeting cuproptosis by FDX1 in acetaminophen-induced liver injury

Abstract

Acetaminophen (APAP) overdose is the most common etiology of acute drug-induced liver injury (DILI) worldwide. Although oxidative stress, apoptosis, and necrosis are well-recognised mechanisms, the involvement of cuproptosis, a newly identified, copper-dependent form of regulated cell death, remains poorly understood. Here, the contribution of cuproptosis to APAP hepatotoxicity is elucidated, identifying ferredoxin 1 (FDX1) as its master regulator. In vivo, the copper chelator ammonium tetrathiomolybdate (TM) markedly attenuated APAP-induced liver damage, as evidenced by decreased serum ALT and AST levels, reduced hepatocyte death, and diminished inflammatory cell infiltration and cytokine release. Transcriptomic mining of GEO datasets revealed significant dysregulation of cuproptosis-associated genes in APAP-exposed hepatocytes, with FDX1 being the most prominently up-regulated. Genetic ablation of FDX1 in mice and FDX1 knockdown in human HepaRG cells both conferred robust protection against APAP-induced lethality, phenocopying the effects of TM. Collectively, our findings establish FDX1-mediated cuproptosis as a critical driver of APAP hepatotoxicity and position copper homeostasis as a tractable therapeutic target for DILI.