CNPY2 controls Macrophage Metabolic Programming and Epigenetic Remodeling 2305267

Abstract Introduction Understanding the metabolic—epigenetic axis in macrophages is critical for human health. In cancer, metabolic and epigenetic reprogramming in tumor-associated macrophages (TAMs) can drive tumor-promoting phenotypes, aiding immune evasion and therapy resistance. However, the regulators linking these processes remain unclear. We previously found that Canopy homolog 2 (CNPY2) promotes chemical hepatocarcinogenesis by regulating macrophage cytokine production, migration, phagocytosis, and cytotoxicity. CNPY2 binds TLR4 and enhances NF-κB—dependent cytokine production in liver macrophages, thereby promoting DEN-induced HCC. Methods To investigate whether CNPY2 influences macrophage immunometabolism and associated chromatin modifications, we generated Cnpy2 knockout mice and used a carcinogen-induced liver cancer mouse model, combining with RNA-seq analysis, qRT-PCR and Western blotting. Results RNA-seq analysis revealed distinct transcriptional changes in metabolic pathways, including de novo lipogenesis, Fatty acid oxidation, and the TCA cycle between wild-type (WT) and Cnpy2-knockout (KO) liver macrophages. Gene-set enrichment analysis further showed altered expression of epigenetic regulators and histone modifying enzymes in Cnpy2-deficient macrophages. We then performed qPCR on selecting metabolic genes, confirming alterations of several RNA-seq-identified targets. Consistent with these findings, protein levels of metabolic enzymes, particularly in acetyl-coA pathway, were significantly increased in Cnpy2 KO liver macrophages. In addition, global histone H3 acetylation was also elevated in Cnpy2 KO liver macrophages compared with WT counterparts. Conclusion Our data suggest that CNPY2 plays a previously unrecognized role in coordinating macrophage metabolic programming and epigenetic regulation. These findings provide an initial framework for understanding how CNPY2 may integrate metabolic cues with chromatin state to shape macrophage function. Funding Source NCI R01CA270166 Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)