Rewired NAD ⁺ metabolism promotes NF- κ B-mediated oxidative stress and disrupts lipid homeostasis in liver fibrosis progression

Chronic liver fibrosis is a major risk factor for the development of hepatocellular carcinoma (HCC), a leading cause of cancer deaths worldwide. However, the underlying molecular mechanisms linking fibrosis to inflammation-associated HCC development remain elusive, hindering early diagnosis and intervention. In this study, we combined unbiased multi-omics analyses, including metabolomics, lipidomics, and transcriptomics, in a mouse model of chemical-induced liver fibrosis and HCC, and integrated publicly available transcriptome data from LX-2 human hepatic stellate cell (HSC) line. Our results revealed a profound rewiring of NAD ⁺ metabolism as a central driver of metabolic disturbance. Multi-omics analysis of the bulk liver tissue showed increased activity of the kynurenine pathway of tryptophan metabolism, boosting the production of NAD ⁺ precursors. Hepatic nicotinamide (NAM) level decreased due to the elevated expression of NAM N -methyltransferase ( Nnmt ) in HSCs. Despite the decreased hepatic NAM, serum NAD ⁺ level was compartmentalized, with increased NAD ⁺ in circulation, and hepatic sirtuins, including Sirt1 , were downregulated to reduce conversion of NAD ⁺ into NAM. This NAM consumption/salvage imbalance derived the disruption of NAD ⁺ homeostasis, triggering the activation of NF- κ B-mediated oxidative stress pathways. Furthermore, lipid dysregulation was evidenced by the domination of NF- κ B regulation to SIRT1/SREBP-controlled lipogenic and cholesterogenic genes, leading to hepatic and serum lipid imbalance. These multi-omics insights elucidate mechanistic links between rewired NAD ⁺ metabolism, inflammation, and lipid dysregulation during fibrosis-HCC progression. Our findings may facilitate the development of novel diagnostic biomarkers and therapeutic targets for this deadly disease.