FTO O-GlcNAcylation downregulates NLRP3 inflammasome and pyroptosis in macrophages

FTO O-GlcNAcylation downregulates NLRP3 inflammasome and pyroptosis in macrophages | 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 Article FTO O-GlcNAcylation downregulates NLRP3 inflammasome and pyroptosis in macrophages Xiao-Lian zhang, Lu Zhang, Min Liu, Yan Xie, Bi-Feng Yuan, Peng Zhiyong, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5654474/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract The fat mass and obesity-associated protein (FTO), a key RNA N6-methyladenosine (m6A) demethylase, has recently emerged as a significant regulator of inflammatory response. However, the specific role and underlying mechanisms of FTO, especially FTO O-GlcNAcylation, in inflammation remain elusive. Here we report a mechanism that O-GlcNAcylation modification controls the biological functions of the FTO protein in macrophage inflammatory response. For the first time, we demonstrate that FTO undergoes O-GlcNAcylation specifically at the Ser95 site. The bacterial endotoxin (lipopolysaccharide, LPS) stimulation enhances FTO O-GlcNAcylation in macrophages through the FOXO1-GFAT2 axis. LPS-enhanced O-GlcNAcylation of FTO promotes TRIM21-mediated FTO ubiquitination degradation and suppresses NLRP3-ASC-Casp1 inflammasome activation and gasdermin D pyroptosis in murine macrophages. FTO O-GlcNAcylation deficiency aggravates S. Typhimurium or LPS-induced sepsis and dextran sulfate sodium (DSS)-induced inflammatory bowel diseases (IBD), while FTO O-GlcNAcylation suppresses the hyperinflammatory phenotype in mice. These findings reveal a novel mechanism that FTO O-GlcNAcylation promotes its degradation and downregulates FTO-mediated inflammatory response. Regulation of FTO O-GlcNAcylation may offer a potential therapeutic strategy for combating endotoxin-induced inflammatory disease and other FTO abnormal expression-associated diseases. Biological sciences/Cell biology/Post-translational modifications/Glycosylation Biological sciences/Chemical biology/Post-translational modifications/Ubiquitylation Biological sciences/Chemical biology/Glycobiology Biological sciences/Immunology/Inflammation/Inflammasome Biological sciences/Immunology/Inflammation/Sepsis O-GlcNAcylation FTO Macrophage Inflammatory response Full Text Additional Declarations There is NO Competing Interest. Supplementary Files SupplementaryInformation.pdf Cite Share Download PDF Status: Posted Version 1 posted 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. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-5654474","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":399592559,"identity":"703b3b84-c285-49cc-b8fd-ac6e4361c86c","order_by":0,"name":"Xiao-Lian 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However, the specific role and underlying mechanisms of FTO, especially FTO O-GlcNAcylation, in inflammation remain elusive. Here we report a mechanism that O-GlcNAcylation modification controls the biological functions of the FTO protein in macrophage inflammatory response. For the first time, we demonstrate that FTO undergoes O-GlcNAcylation specifically at the Ser95 site. The bacterial endotoxin (lipopolysaccharide, LPS) stimulation enhances FTO O-GlcNAcylation in macrophages through the FOXO1-GFAT2 axis. LPS-enhanced O-GlcNAcylation of FTO promotes TRIM21-mediated FTO ubiquitination degradation and suppresses NLRP3-ASC-Casp1 inflammasome activation and gasdermin D pyroptosis in murine macrophages. FTO O-GlcNAcylation deficiency aggravates S. Typhimurium or LPS-induced sepsis and dextran sulfate sodium (DSS)-induced inflammatory bowel diseases (IBD), while FTO O-GlcNAcylation suppresses the hyperinflammatory phenotype in mice. 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