Multiscale identification of DNASE1L3 as a key target in MASLD progression to hepatocellular carcinoma.

Background: The rising incidence of hepatocellular carcinoma (HCC) linked to metabolic dysfunction-associated steatotic liver disease (MASLD) underscores the need to identify key drivers of malignant transformation. This study aimed to discover critical genes governing MASLD-to-HCC progression for early intervention.

Methods: Bulk and single-cell transcriptomic data were analyzed using pseudotime algorithms (Mfuzz, Monocle3) to pinpoint progression-related genes. The scissor algorithm identified target cells, and diagnostic models were developed to validate MASLD-associated genes. Functional validation included siRNA knockdown and plasmid-driven overexpression of DNASE1L3 in MASLD and MASLD-HCC cell models. Protein and mRNA expression was assessed via Western blotting and RTqPCR. Lipid accumulation (Oil Red O staining) and proliferation (CCK8 assays) were evaluated post-intervention.

Results: DNASE1L3 emerged as a pivotal gene, with marked downregulation in MASLD and HCC models. Overexpression suppressed HCC cell proliferation while enhancing lipid accumulation, whereas knockdown showed no significant effects.

Conclusions: DNASE1L3 is a potential therapeutic target in MASLD-driven HCC, regulating malignant progression through dual modulation of proliferation and lipid metabolism. These findings provide mechanistic insights for future interventions.