FTO O-GlcNAcylation induces inactive form Pyrin expression, which suppresses NLRP3 inflammasome activation during LPS stimulation

FTO O-GlcNAcylation induces inactive form Pyrin expression, which suppresses NLRP3 inflammasome activation during LPS stimulation | 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 induces inactive form Pyrin expression, which suppresses NLRP3 inflammasome activation during LPS stimulation Xiao-Lian Zhang, Lu Zhang, Min Liu, Yan Xie, Bi-Feng Yuan, Zhiyong Peng, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5408730/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), as a key RNA N6-methyladenosine (m6A) demethylase, has been recently highlighted in modulating inflammatory response. However, the role and underlying mechanisms of FTO, especially FTO O-GlcNAcylation, in LPS-induced inflammasome remain elusive. Here we uncover that FTO undergoes O-GlcNAcylation specifically at the Ser95 site. LPS-enhanced O-GlcNAcylation of FTO promotes TRIM21-mediated FTO ubiquitination degradation, subsequently increasing m6A-modified Mefv (encoding Pyrin) expression in macrophages. LPS stimulation or FTO O-GlcNAcylation increases inactive form Pyrin expression and can not cause Pyrin-ASC-Casp1 inflammasome activation. The Pyrin competitively blocks NLRP3-ASC-Casp1 inflammasome assembly and activation through the Pyrin-PYD domain, reduces gasdermin D (GSDMD) pyroptosis and restrains TNF-α/IL-1β/IL-6 production. FTO O-GlcNAcylation-deficiency or Pyrin knockdown aggravates the S. Typhimurium or LPS-induced sepsis and dextran sulfate sodium (DSS)-induced inflammatory bowel diseases (IBD). Our findings clarify that there is a regulatory network among protein O-GlcNAcylation, m6A modification, and inflammasome activation. FTO O-GlcNAcylation or Pyrin acts as a negative regulator of NLPR3-ASC-Casp1 inflammasome. The Pyrin plays an anti-inflammatory role in LPS-treated macrophages. Enhancement of FTO O-GlcNAcylation or inactive form Pyrin may offer a potential therapeutic strategy for combating endotoxin-induced NLRP3-mediated inflammatory responses and sepsis. Biological sciences/Immunology/Inflammation/Inflammasome Biological sciences/Cell biology/Glycobiology Biological sciences/Molecular biology/Post-translational modifications/Glycosylation O-GlcNAcylation FTO Inflammation Pyrin (Mefv) m6A 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-5408730","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":377386105,"identity":"e5d59617-78e0-42a9-a4c5-a6d427a1dd1d","order_by":0,"name":"Xiao-Lian Zhang","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABBUlEQVRIiWNgGAWjYDACCRjJztjAwFDBkIAQJKiFGaTlDPFagIAZiBnbiNAiP7v52cOvbRby/MzMjZ8L5x3OMzjAfPA2D4NdHi4tjHOOmRvLnJEwnNnM2Cw9c9vhYoMDbMnWPAzJxbi0MEskmElLVEgwbjjM2CDNu+124oYDPGbSPAwHEhtwaGGTSP8mLWEgYQ/U0vybdw5IC/83vFp4JHLMJD9USCQCtbRJ8zaAbWHDq0VCIqdMmuGMRDLQL23WPMf+J848zGZsOccgGacW+Rnp2yR/ttXZ9rO3P77NU5OW2He8+eGNNxV2OLWAg4AHlQsiDPCoBwLGH/jlR8EoGAWjYKQDAF9+UoUAl4krAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0002-8283-9381","institution":"Wuhan University Taikang Medical School (School of Basic Medical Sciences)","correspondingAuthor":true,"prefix":"","firstName":"Xiao-Lian","middleName":"","lastName":"Zhang","suffix":""},{"id":377386106,"identity":"938d2105-911c-42e6-9398-8fc0ffb67bbb","order_by":1,"name":"Lu Zhang","email":"","orcid":"","institution":"Department of Immunology Wuhan University Taikang Medical School (School of Basic Medical Sciences)","correspondingAuthor":false,"prefix":"","firstName":"Lu","middleName":"","lastName":"Zhang","suffix":""},{"id":377386107,"identity":"f2680147-d2d2-4b63-abb1-7bca936fbf98","order_by":2,"name":"Min Liu","email":"","orcid":"","institution":"Wuhan University School of Medicine","correspondingAuthor":false,"prefix":"","firstName":"Min","middleName":"","lastName":"Liu","suffix":""},{"id":377386108,"identity":"1bc86ab3-3be3-4d08-acb6-984a172f8c15","order_by":3,"name":"Yan Xie","email":"","orcid":"","institution":"Wuhan University","correspondingAuthor":false,"prefix":"","firstName":"Yan","middleName":"","lastName":"Xie","suffix":""},{"id":377386109,"identity":"6ff81377-68e0-49c3-a3be-f3eccb65948e","order_by":4,"name":"Bi-Feng Yuan","email":"","orcid":"","institution":"School of Public Health, Wuhan University, Wuhan","correspondingAuthor":false,"prefix":"","firstName":"Bi-Feng","middleName":"","lastName":"Yuan","suffix":""},{"id":377386110,"identity":"b43d58fd-d07c-4526-8356-04d4c91d6fac","order_by":5,"name":"Zhiyong Peng","email":"","orcid":"","institution":"Zhongnan Hospital of Wuhan University, Wuhan","correspondingAuthor":false,"prefix":"","firstName":"Zhiyong","middleName":"","lastName":"Peng","suffix":""},{"id":377386111,"identity":"387e8f78-9e8c-4680-a921-e6c62c28e6fa","order_by":6,"name":"Jun Xiong","email":"","orcid":"","institution":"School of Public Health, Wuhan University, Wuhan","correspondingAuthor":false,"prefix":"","firstName":"Jun","middleName":"","lastName":"Xiong","suffix":""}],"badges":[],"createdAt":"2024-11-07 09:45:16","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5408730/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5408730/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":69754412,"identity":"c96d0d08-7450-49b7-98f5-792049705da6","added_by":"auto","created_at":"2024-11-25 02:59:20","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":9451447,"visible":true,"origin":"","legend":"","description":"","filename":"ManuscriptforNatureImmunology.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5408730/v1_covered_885c9bf2-cdd1-4d55-b634-d94b413d162d.pdf"},{"id":69752879,"identity":"45e4ea2d-1eb9-4736-acdf-9c201619b6f0","added_by":"auto","created_at":"2024-11-25 02:35:04","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":2936102,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cbr\u003e\u003c/p\u003e","description":"","filename":"SupplementaryInformation.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5408730/v1/d8ba8cf395eb4a652e2b9700.pdf"}],"financialInterests":"There is \u003cb\u003eNO\u003c/b\u003e Competing Interest.","formattedTitle":"FTO O-GlcNAcylation induces inactive form Pyrin expression, which suppresses NLRP3 inflammasome activation during LPS stimulation","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"O-GlcNAcylation, FTO, Inflammation, Pyrin (Mefv), m6A","lastPublishedDoi":"10.21203/rs.3.rs-5408730/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5408730/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"The fat mass and obesity-associated protein (FTO), as a key RNA N6-methyladenosine (m6A) demethylase, has been recently highlighted in modulating inflammatory response. However, the role and underlying mechanisms of FTO, especially FTO O-GlcNAcylation, in LPS-induced inflammasome remain elusive. Here we uncover that FTO undergoes O-GlcNAcylation specifically at the Ser95 site. LPS-enhanced O-GlcNAcylation of FTO promotes TRIM21-mediated FTO ubiquitination degradation, subsequently increasing m6A-modified Mefv (encoding Pyrin) expression in macrophages. LPS stimulation or FTO O-GlcNAcylation increases inactive form Pyrin expression and can not cause Pyrin-ASC-Casp1 inflammasome activation. The Pyrin competitively blocks NLRP3-ASC-Casp1 inflammasome assembly and activation through the Pyrin-PYD domain, reduces gasdermin D (GSDMD) pyroptosis and restrains TNF-α/IL-1β/IL-6 production. FTO O-GlcNAcylation-deficiency or Pyrin knockdown aggravates the S. Typhimurium or LPS-induced sepsis and dextran sulfate sodium (DSS)-induced inflammatory bowel diseases (IBD). Our findings clarify that there is a regulatory network among protein O-GlcNAcylation, m6A modification, and inflammasome activation. FTO O-GlcNAcylation or Pyrin acts as a negative regulator of NLPR3-ASC-Casp1 inflammasome. The Pyrin plays an anti-inflammatory role in LPS-treated macrophages. Enhancement of FTO O-GlcNAcylation or inactive form Pyrin may offer a potential therapeutic strategy for combating endotoxin-induced NLRP3-mediated inflammatory responses and sepsis.","manuscriptTitle":"FTO O-GlcNAcylation induces inactive form Pyrin expression, which suppresses NLRP3 inflammasome activation during LPS stimulation","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-11-25 02:34:59","doi":"10.21203/rs.3.rs-5408730/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"ca547d44-6711-4acd-b139-b4c484d7ed33","owner":[],"postedDate":"November 25th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":40178147,"name":"Biological sciences/Immunology/Inflammation/Inflammasome"},{"id":40178148,"name":"Biological sciences/Cell biology/Glycobiology"},{"id":40178149,"name":"Biological sciences/Molecular biology/Post-translational modifications/Glycosylation"}],"tags":[],"updatedAt":"2024-11-25T02:35:02+00:00","versionOfRecord":[],"versionCreatedAt":"2024-11-25 02:34:59","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5408730","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5408730","identity":"rs-5408730","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}