Multi-Omics Characterization of an Alcohol-Induced Hepatocellular Carcinoma Mouse Model

Hepatocellular carcinoma (HCC) is a major global health problem ranking as the sixth most commonly diagnosed cancer , and the fourth leading cause of cancer-related deaths worldwide. Although the incidence of viral infection-mediated HCC has decreased in recent years, the incidence of alcohol- and metabolic dysfunction-associated HCC has increased, which is driven by changes in lifestyle and diet. Excessive alcohol consumption contributes to advanced liver disease, including liver fibrosis, cirrhosis, and HCC. Despite its clinical relevance, there is a lack of suitable animal models that adequately reflect the pathophysiological features of alcohol-associated HCC in humans. To address these limitations, we established a novel mouse model of alcohol-associated HCC through the combined administration of N-diethylnitrosamine (DEN) and carbon tetrachloride (CCl₄), followed by alcohol-containing Lieber-DeCarli diet. The results indicated that chronic alcohol administration in the presence of DEN and CCl₄ significantly accelerated HCC development, which was characterized by increased oxidative stress, inflammation, and severe fibrosis. Furthermore, we found that chronic ethanol exposure disrupted hepatic immunity, which was characterized by natural killer (NK)/NKT cell depletion, increased PD-1⁺CD8⁺ cells, reduced cytotoxicity, and elevated inflammation. We also observed significant alterations in the gut microbiome following chronic alcohol consumption. Such immunological and microbiome alternations fostered an immunosuppressive microenvironment that accelerates HCC progression. In conclusion, our newly developed mouse model effectively induced liver tumorigenesis within a relatively short time and recapitulated the clinical and pathological features of alcohol-associated HCC. This model represents a valuable tool for studying the mechanisms of alcohol-related liver cancer and for developing targeted therapies.