Loss of hepatic DRP1 exacerbates alcoholic hepatitis by inducing megamitochondria and mitochondrial maladaptation

<jats:sec> <jats:title>Background and Aims:</jats:title> <jats:p>Increased megamitochondria formation and impaired mitophagy in hepatocytes have been linked to the pathogenesis of alcohol‐associated liver disease (ALD). This study aims to determine the mechanisms by which alcohol consumption increases megamitochondria formation in the pathogenesis of ALD.</jats:p> </jats:sec> <jats:sec> <jats:title>Approach and Results:</jats:title> <jats:p>Human alcoholic hepatitis (AH) liver samples were used for electron microscopy, histology, and biochemical analysis. Liver‐specific dynamin‐related protein 1 (DRP1; gene name <jats:italic toggle="yes">DNM1L, an essential gene regulating mitochondria fission</jats:italic>) knockout (L‐DRP1 KO) mice and wild‐type mice were subjected to chronic plus binge alcohol feeding. Both human AH and alcohol‐fed mice had decreased hepatic DRP1 with increased accumulation of hepatic megamitochondria. Mechanistic studies revealed that alcohol feeding decreased DRP1 by impairing transcription factor EB–mediated induction of <jats:italic toggle="yes">DNM1L</jats:italic>. L‐DRP1 KO mice had increased megamitochondria and decreased mitophagy with increased liver injury and inflammation, which were further exacerbated by alcohol feeding. Seahorse flux and unbiased metabolomics analysis showed alcohol intake increased mitochondria oxygen consumption and hepatic nicotinamide adenine dinucleotide (NAD<jats:sup>+</jats:sup>), acylcarnitine, and ketone levels, which were attenuated in L‐DRP1 KO mice, suggesting that loss of hepatic DRP1 leads to maladaptation to alcohol‐induced metabolic stress. RNA‐sequencing and real‐time quantitative PCR analysis revealed increased gene expression of the cGAS–stimulator of interferon genes (STING)–interferon pathway in L‐DRP1 KO mice regardless of alcohol feeding. Alcohol‐fed L‐DRP1 KO mice had increased cytosolic mtDNA and mitochondrial dysfunction leading to increased activation of cGAS‐STING‐interferon signaling pathways and liver injury.</jats:p> </jats:sec> <jats:sec> <jats:title>Conclusion:</jats:title> <jats:p>Alcohol consumption decreases hepatic DRP1 resulting in increased megamitochondria and mitochondrial maladaptation that promotes AH by mitochondria‐mediated inflammation and cell injury.</jats:p> </jats:sec>