IFIT3 Knockdown Attenuates Pressure-Overload-Induced Cardiac Inflammation and Remodeling Through a JNK/H3K9 Lactylation-Associated Mechanism

IFIT3 Knockdown Attenuates Pressure-Overload-Induced Cardiac Inflammation and Remodeling Through a JNK/H3K9 Lactylation-Associated Mechanism | 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 IFIT3 Knockdown Attenuates Pressure-Overload-Induced Cardiac Inflammation and Remodeling Through a JNK/H3K9 Lactylation-Associated Mechanism Qingyi Luo, Chunrui Zhou, Yuying Wang, Li Zhao, Jian Zhang, Yunchuan Ding, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9292460/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 Aims: Heart failure (HF) is characterized by sustained inflammation and adverse cardiac remodeling, yet the mechanisms underlying macrophage-associated inflammatory activation remain incompletely understood. This study investigated whether the interferon-induced protein IFIT3 contributes to pressure-overload-induced HF and explored the potential involvement of JNK signaling and histone H3 lysine 9 lactylation (H3K9la). Methods and Results: We performed integrated bioinformatics analyses of publicly available single-cell RNA sequencing data from human failing hearts (GEO: GSE145154) and identified IFIT3 as a macrophage-enriched hub gene. In vivo , we used AAV-mediated IFIT3 knockdown in a transverse aortic constriction (TAC) mouse model. In vitro , we performed siRNA-mediated IFIT3 silencing in RAW264.7 macrophage-like cells and rescue treatments with anisomycin and sodium lactate. Single-cell transcriptomic analysis revealed that IFIT3 was significantly upregulated in macrophages from failing human hearts. In TAC-induced heart failure in mice, myocardial IFIT3 expression was increased and showed partial colocalization with CD68-positive cells. AAV-mediated IFIT3 knockdown improved cardiac function, reduced inflammatory cytokine production, and attenuated myocardial fibrosis in vivo . In RAW264.7 macrophage-like cells, IFIT3 silencing suppressed lipopolysaccharide-induced inflammatory cytokine secretion and reduced JNK phosphorylation and H3K9la levels. Rescue experiments showed that anisomycin partially restored p-JNK, H3K9la, and inflammatory cytokine expression, whereas sodium lactate supplementation partially rescued H3K9la levels and inflammatory responses in IFIT3 -silenced cells. Conclusion: These findings suggest that IFIT3 contributes to pressure-overload-induced cardiac inflammation and remodeling and that this effect is associated with JNK activation and H3K9 lactylation. IFIT3 may represent a potential target for immunoinflammatory modulation in heart failure. Heart failure Macrophages IFIT3 JNK signaling pathway H3K9 lactylation Inflammation Full Text Additional Declarations No competing interests reported. Supplementary Files Additionalfile1.docx 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-9292460","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":619997564,"identity":"f342e78f-9973-482d-a9b9-bb1ce6d48c18","order_by":0,"name":"Qingyi Luo","email":"","orcid":"","institution":"Yan'an Hospital Affiliated To Kunming Medical University","correspondingAuthor":false,"prefix":"","firstName":"Qingyi","middleName":"","lastName":"Luo","suffix":""},{"id":619997565,"identity":"4766b866-a9b4-44de-9601-656e48afda0d","order_by":1,"name":"Chunrui Zhou","email":"","orcid":"","institution":"Yan'an Hospital Affiliated To Kunming 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Mechanism","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"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":"Heart failure, Macrophages, IFIT3, JNK signaling pathway, H3K9 lactylation, Inflammation","lastPublishedDoi":"10.21203/rs.3.rs-9292460/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-9292460/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eAims:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eHeart failure (HF) is characterized by sustained inflammation and adverse cardiac remodeling, yet the mechanisms underlying macrophage-associated inflammatory activation remain incompletely understood. This study investigated whether the interferon-induced protein \u003cem\u003eIFIT3\u003c/em\u003e contributes to pressure-overload-induced HF and explored the potential involvement of JNK signaling and histone H3 lysine 9 lactylation (H3K9la).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods and Results:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe performed integrated bioinformatics analyses of publicly available single-cell RNA sequencing data from human failing hearts (GEO: GSE145154) and identified \u003cem\u003eIFIT3\u003c/em\u003eas a macrophage-enriched hub gene. \u003cem\u003eIn vivo\u003c/em\u003e, we used AAV-mediated \u003cem\u003eIFIT3\u003c/em\u003eknockdown in a transverse aortic constriction (TAC) mouse model. \u003cem\u003eIn vitro\u003c/em\u003e, we performed siRNA-mediated \u003cem\u003eIFIT3\u003c/em\u003e silencing in RAW264.7 macrophage-like cells and rescue treatments with anisomycin and sodium lactate.\u003c/p\u003e\n\u003cp\u003eSingle-cell transcriptomic analysis revealed that \u003cem\u003eIFIT3\u003c/em\u003e was significantly upregulated in macrophages from failing human hearts. In TAC-induced heart failure in mice, myocardial \u003cem\u003eIFIT3\u003c/em\u003e expression was increased and showed partial colocalization with CD68-positive cells. AAV-mediated \u003cem\u003eIFIT3\u003c/em\u003eknockdown improved cardiac function, reduced inflammatory cytokine production, and attenuated myocardial fibrosis \u003cem\u003ein vivo\u003c/em\u003e. In RAW264.7 macrophage-like cells, \u003cem\u003eIFIT3\u003c/em\u003e silencing suppressed lipopolysaccharide-induced inflammatory cytokine secretion and reduced JNK phosphorylation and H3K9la levels. Rescue experiments showed that anisomycin partially restored p-JNK, H3K9la, and inflammatory cytokine expression, whereas sodium lactate supplementation partially rescued H3K9la levels and inflammatory responses in \u003cem\u003eIFIT3\u003c/em\u003e-silenced cells.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThese findings suggest that \u003cem\u003eIFIT3\u003c/em\u003e contributes to pressure-overload-induced cardiac inflammation and remodeling and that this effect is associated with JNK activation and H3K9 lactylation. \u003cem\u003eIFIT3\u003c/em\u003e may represent a potential target for immunoinflammatory modulation in heart failure.\u003c/p\u003e","manuscriptTitle":"IFIT3 Knockdown Attenuates Pressure-Overload-Induced Cardiac Inflammation and Remodeling Through a JNK/H3K9 Lactylation-Associated Mechanism","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-04-11 00:58:56","doi":"10.21203/rs.3.rs-9292460/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":"319a7064-98cf-4bf9-aade-139862717fbe","owner":[],"postedDate":"April 11th, 2026","published":true,"recentEditorialEvents":[{"type":"decision","content":"Rejected","date":"2026-05-14T17:16:16+00:00","index":"","fulltext":""}],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-05-14T17:25:13+00:00","versionOfRecord":[],"versionCreatedAt":"2026-04-11 00:58:56","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-9292460","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-9292460","identity":"rs-9292460","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}