Mouse embryonic stem cells exhibit cancer-like DNA methylation landscapes upon DNMT1 overexpression

Mouse embryonic stem cells exhibit cancer-like DNA methylation landscapes upon DNMT1 overexpression | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Mouse embryonic stem cells exhibit cancer-like DNA methylation landscapes upon DNMT1 overexpression Elizabeth Elder, Anthony Lemieux, Serge McGraw This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9052961/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 10 You are reading this latest preprint version Abstract DNA methyltransferase 1 (DNMT1) overexpression is a recurrent feature and an emerging therapeutic target in cancer, yet its direct impact on DNA methylation regulation remains poorly defined. Using enzymatic methyl sequencing, we show that DNMT1 overexpression in mouse embryonic stem cells leads to DNA methylation landscapes strikingly reminiscent of cancer methylomes, characterized by global hypomethylation, focal hypermethylation in promoters and CpG islands, and increased CpG-to-CpG variability. Differentially methylated regions in promoters correlate with altered gene expression and intersect with binding sites of key transcriptional regulators, pointing to potential underlying mechanisms of DNMT1 overexpression. We also show that DNMT3A and DNMT3B levels become lowered, suggesting that the broader DNA methylation machinery is perturbed when DNMT1 is in excess. Finally, we reveal that hypermethylation is partially retained following acute DNMT1 depletion and conserved across diverse human cancer methylomes, highlighting the translational relevance of this study for understanding the pathological mechanisms of DNMT1 overexpression in cancer and the limitations of DNMT1-targeted therapeutic strategies. Full Text Additional Declarations No competing interests reported. Supplementary Files SupplementarymaterialsElderetal2026.pdf SupplementarydataElderetal2026.xlsx Cite Share Download PDF Status: Under Review Version 1 posted Reviews received at journal 27 Apr, 2026 Reviews received at journal 20 Apr, 2026 Reviews received at journal 09 Apr, 2026 Reviewers agreed at journal 03 Apr, 2026 Reviewers agreed at journal 29 Mar, 2026 Reviewers agreed at journal 29 Mar, 2026 Reviewers invited by journal 29 Mar, 2026 Editor assigned by journal 16 Mar, 2026 Submission checks completed at journal 09 Mar, 2026 First submitted to journal 06 Mar, 2026 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-9052961","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":614872812,"identity":"ebbf3619-01f5-4f39-8869-40def26eb8b3","order_by":0,"name":"Elizabeth Elder","email":"","orcid":"","institution":"University of Montreal","correspondingAuthor":false,"prefix":"","firstName":"Elizabeth","middleName":"","lastName":"Elder","suffix":""},{"id":614872813,"identity":"822549e8-8975-480a-9ccb-ad0d308baaae","order_by":1,"name":"Anthony Lemieux","email":"","orcid":"","institution":"Azrieli Research Centre of Sainte-Justine University 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