Abstract A031: Multi-omics profiling reveals cancer-relevant gut–liver axis alterations during MASL to MASH transition

Abstract Background: Metabolic dysfunction-associated steatotic liver disease (MASLD) and its advanced form, metabolic dysfunction-associated steatohepatitis (MASH), are global health concerns driven by the obesity epidemic. Beyond their roles in cirrhosis and hepatocellular carcinoma (HCC), emerging evidence links MASLD and MASH to increased colorectal neoplasia and colorectal cancer (CRC). However, the molecular and cellular mechanisms by which steatotic liver disease fosters a pro-tumorigenic gut-liver axis remain poorly defined. Methods: Male C57BL/6NTac mice were fed a high-fat diet for 16 weeks (MASLD) or 29 weeks (MASH). We performed untargeted metabolomics and lipidomics in liver, colon, and stool; quantified circulating enteroendocrine (EEC) hormones and cytokines; examined EEC lineage in colonic organoids; and assessed hepatic serotonin and fibrosis by immunofluorescence. Results: Histology confirmed progression from MASLD to MASH. Untargeted metabolomics revealed consistent elevation of cholic acid across tissues and disease stages; mapping analyses linked metabolite changes to CRC-associated pathways. Lipidomics showed stage-specific remodeling: MASLD favored lipotoxic species (free fatty acids, diacylglycerols, LPC/LPE) associated with insulin resistance and oxidative stress, whereas MASH accumulated acylcarnitines, markers of β-oxidation overload tied to hepatocarcinogenesis. In early disease (MASLD), circulating GLP-1, GIP, and PYY were elevated and correlated with colonic cholic acid, while MASH lost this incretin response but exhibited higher TNFα and IL-6. Colonic organoids showed increased CHGA staining, suggesting EEC expansion that was lost in MASH. In the liver, serotonin accumulated and partially colocalized with fibronectin, consistent with a fibrotic niche implicated in tumor initiation. Conclusions: These findings reveal progressive metabolic and hormonal reprogramming from MASLD to MASH that converges on cancer-relevant pathways. Rising cholic acid, early incretin activation followed by EEC dysfunction, and serotonergic fibrosis together establish a pro-tumorigenic gut-liver axis. This integrative framework highlights new mechanistic links between steatotic liver disease and both CRC and HCC, offering potential avenues for prevention and therapy. Citation Format: Jaclyn A. Rivas, Alexandria C. Murphy, Praveena Prasad, Siem S. Goitom, Aaron S. Romero, Crystal M . Enriquez, Brianna B. Maes, Prithvi R. Akepati, Marcus A. Garcia, Fredine T. Lauer, Rama R. Gullapalli, Kristen M. Gonzales, Jessica M . Gross, Jing Pu, Shuguang Leng, Julie G. In, Melanie R. McReynolds, Eliseo F. Castillo. Multi-omics profiling reveals cancer-relevant gut–liver axis alterations during MASL to MASH transition [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Cancer Evolution: The Dynamics of Progression and Persistence; 2025 Dec 4-6; Albuquerque, NM. Philadelphia (PA): AACR; Cancer Res 2025;85(23_Suppl):Abstract nr A031.