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Aberrant DNA methylation, which is primarily maintained by DNA methyltransferase 1 (DNMT1), has been linked to metabolic dysregulation; however, its contribution to MASLD pathogenesis remains poorly defined. This study aimed to elucidate the role of DNMT1-mediated methylation in transcriptional regulation during MASLD progression and to determine whether DNMT1 inhibition can reverse disease-associated epigenetic and transcriptional alterations.Methods: We conducted integrated analyses of the liver transcriptome (n=131) and DNA methylome (n=106) of patients with biopsy-proven MASLD. We evaluated the effect of DNMT1 inhibition with 5-aza-4\u2032-thio-2\u2032-deoxycytidine (Aza-TdC) on a diet-induced MASLD mouse model. Multiomics approaches, including DNA methylome profiling, lipidomics, RNA sequencing, and chromatin immunoprecipitation sequencing, were applied to elucidate the role of DNMT1-mediated DNA methylation in regulating pathogenic gene expression.Results: DNA methylome profiling revealed increased methylation variability associated with increased DNMT1 expression in MASLD patients. DNMT1 inhibition ameliorated dysregulated lipid metabolism by reducing hepatic triacylglycerol accumulation and inflammation. Aza-TdC treatment partially reversed MASLD-related hypermethylation of hepatocyte nuclear factor 4 alpha (HNF4\u03b1)- and peroxisome proliferator-activated receptor alpha (PPAR\u03b1)-regulated genes, restoring their transcriptional activity. Notably, Aza-TdC reactivated the gluconeogenic enzyme-encoding gene &lt;i&gt;phosphoenolpyruvate carboxykinase 1&lt;\/i&gt; (&lt;i&gt;PCK1&lt;\/i&gt;), which was hypermethylated and transcriptionally repressed in MASLD. Targeted DNA methylation of the PCK1 promoter using CRISPRoff confirmed the direct epigenetic regulation of PCK1 expression.Conclusions: Targeting DNMT1 may mitigate lipid dysregulation and inflammation by reversing hypermethylation and restoring HNF4\u03b1- and PPAR\u03b1-dependent gene transcription, highlighting DNMT1 as a potential therapeutic target for MASLD.<\/jats:p>","DOI":"10.3350\/cmh.2025.1099","type":"journal-article","created":{"date-parts":[[2026,1,27]],"date-time":"2026-01-27T23:46:37Z","timestamp":1769557597000},"page":"1201-1224","update-policy":"https:\/\/doi.org\/10.3350\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["DNMT1 facilitates the progression of metabolic dysfunction-associated steatotic liver disease by impeding transcription mediated by HNF4\u03b1 and PPAR\u03b1"],"prefix":"10.3350","volume":"32","author":[{"given":"Hyun Ahm","family":"Sohn","sequence":"first","affiliation":[{"name":"Aging Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hanyong","family":"Go","sequence":"additional","affiliation":[{"name":"Aging Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, Korea"},{"name":"Department of Functional Genomics, KRIBB School of Bioscience, Korea University of Science and Technology (UST), Daejeon, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tae Hyeon","family":"An","sequence":"additional","affiliation":[{"name":"Metabolic Regulation Research Center, KRIBB, Daejeon, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jun Min","family":"Lee","sequence":"additional","affiliation":[{"name":"Aging Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, Korea"},{"name":"Department of Functional Genomics, KRIBB School of Bioscience, Korea University of Science and Technology (UST), Daejeon, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hee-Jin","family":"Kim","sequence":"additional","affiliation":[{"name":"Aging Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Keeok","family":"Haam","sequence":"additional","affiliation":[{"name":"Korea Bioinformation Center (KOBIC), KRIBB, Daejeon, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amal","family":"Magdy","sequence":"additional","affiliation":[{"name":"Aging Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, Korea"},{"name":"Department of Functional Genomics, KRIBB School of Bioscience, Korea University of Science and Technology (UST), Daejeon, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hyo-Jung","family":"Jung","sequence":"additional","affiliation":[{"name":"Aging Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yang-Ji","family":"Shin","sequence":"additional","affiliation":[{"name":"Smart Farm Research Center, Korea Institute of Science and Technology (KIST), Gangneung Institute of Natural Products, Gangneung, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hyun Jung","family":"Lim","sequence":"additional","affiliation":[{"name":"Aging Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, Korea"},{"name":"Department of Functional Genomics, KRIBB School of Bioscience, Korea University of Science and Technology (UST), Daejeon, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yujin","family":"Jeong","sequence":"additional","affiliation":[{"name":"Aging Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, Korea"},{"name":"Department of Functional Genomics, KRIBB School of Bioscience, Korea University of Science and Technology (UST), Daejeon, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yejin","family":"Bae","sequence":"additional","affiliation":[{"name":"Integrated Metabolomics Research Group, Metropolitan Seoul Center, Korea Basic Science Institute, Seoul, Korea"},{"name":"Department of Chemistry, Sungkyunkwan University, Suwon, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Youngae","family":"Jung","sequence":"additional","affiliation":[{"name":"Integrated Metabolomics Research Group, Metropolitan Seoul Center, Korea Basic Science Institute, Seoul, 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