Spatial transcriptomic profiling of gene expression alterations following ACBP/DBI inhibition in a MASH-Driven HCC mouse model
We identified ACBP/DBI as a critical regulator in the pathogenesis of hepatocellular carcinoma (HCC), with its inhibition significantly impairing hepatocarcinogenesis in MASH-driven HCC models. To further elucidate the underlying transcriptional mechanisms, we performed spatial transcriptomic analyses using formalin-fixed, paraffin-embedded (FFPE) liver tissues from the following two experimental models: (i) Western diet (WD) and CCl4-treated mice with tamoxifen-inducible ACBP/DBI knockout (Dbi?/? mice), with Dbi?/? littermates serving as controls, after a total of 27 weeks; (ii) WD and CCl4-treated mice immunized with either keyhole limpet hemocyanin (KLH) alone or KLH-ACBP conjugate, after a total of 34 weeks. Overall design: (i) In the WD+CCl4 model, Dbi?/? and Dbi?/? mice received intraperitoneal (i.p.) injections of tamoxifen at a dose of 75 mg/kg/day for five consecutive days to induce gene deletion. Three days after the final tamoxifen injection, mice were switched to a Western diet supplemented with high-sugar water (23.1 g/L D-fructose and 18.9 g/L D-glucose) and received weekly i.p. injections of carbon tetrachloride (CCl4) for 26 weeks. (ii) In the WD+CCl4 model with KLH/KLH-ACBP immunization, five-week-old mice were vaccinated intraperitoneally with a mixture of KLH/Montanide or KLH-ACBP/Montanide on days 0 (30 µg), 7 (30 µg), 14 (30 µg), and 21 (10 µg) to induce the generation of anti-ACBP autoantibodies. Following vaccination, mice were placed on a Western diet supplemented with high-sugar water (23.1 g/L D-fructose and 18.9 g/L D-glucose) and received weekly i.p. injections of CCl4 for 30 weeks. At the end of the experimental period, liver tissues were collected, placed into cassettes, fixed in 4% paraformaldehyde for 24 hours, and subsequently transferred to 70% ethanol. Following standard dehydration and processing steps, tissues were embedded in paraffin for spatial transcriptomic analyses. All mice used in these experiments were male and of C57BL/6 genetic background.