Targeted inhibition NCOA4-mediated ferritinophagy limits ferroptosis to ameliorate alcohol-induced liver disease in vivo and in vitro

Alcohol-induced liver disease (ALD) is a serious liver disease resulting from prolonged and excessive alcohol consumption, with significant morbidity and mortality and closely relate to iron accumulation and ferroptosis. Ferritinophagy is the process of autophagic degradation of ferritin and results in labile iron accumulation and ferroptosis. However, whether ferritinophagy involves in ALD and ferroptosis remains unexplored. Here, we demonstrated the involvement of ferroptosis in ALD by modulating nuclear receptor coactivator 4 (NCOA4)-mediated ferritinophagy. Utilizing both in vivo and in vitro models, we observed activation of autophagy and ferroptosis following alcohol administration. Intriguingly, we found that alcohol-induced ferroptosis is autophagy-dependent, because autophagy inhibitor 3-methyladenine (3-MA, 30 mg/kg bw in vivo, 1 mmol/L in vitro, respectively) profoundly mitigated alcohol-induced ferroptosis. We further demonstrated that alcohol-induced ferritinophagy was derived by iron overload, by showing that iron chelator deferoxamine (DFO, 100 mg/kg bw in vivo, 10 μmol/L in vitro, respectively) can abolish alcohol-induced ferritinophagy and ferroptosis. Moreover, we observed an upregulation of NCOA4 expression, and knockdown of NCOA4 significantly reversed alcohol-induced disturbances in iron metabolism and subsequently blocking ferroptosis. Our findings suggest a connection between ferritinophagy and alcohol-induced ferroptosis in mouse liver and HepG2 cells, highlighting NCOA4-mediated ferritinophagy as a novel mechanism of ALD. This study enhances our understanding of the molecular mechanisms underlying ALD, and targeting ferritinophagy may provide new strategies for preventing or treating ALD.