Pathogenic Role of Cytoplasmic IKKα in Meibomian Gland Dysfunction and Dry Eye Disease

Pathogenic Role of Cytoplasmic IKKα in Meibomian Gland Dysfunction and Dry Eye Disease | 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 Pathogenic Role of Cytoplasmic IKKα in Meibomian Gland Dysfunction and Dry Eye Disease M. Llanos Casanova, Verónica García-García, Angustias Page, Alejandra Conde, and 8 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9300712/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted You are reading this latest preprint version Abstract Dry eye disease (DED) is a multifactorial ocular surface disorder affecting over 20% of adults over 40, with higher prevalence in the elderly. Meibomian gland dysfuncton(MGD) is its leading cause. The meibomian glands (MGs) are holocrine sebaceous glands composed of acini, where basal cells differentiate into meibocytes that produce meibum, a lipid-rich substance essential for tear film stability and preventing evaporation. Inflammation is central to DED/MGD pathophysiology, driving a self-perpetuating cycle that disrupts innate and adaptive immunity and promotes chronicity. Overactivation of NF-κB -a key mediator of inflammation and aging- on the ocular surface likely plays a critical role in DED/MGD, though the mechanisms of its activation remain unclear, potentially limiting treatment efficacy. Here, we demonstrate that C-IKKα transgenic mice, which express exogenous IKKα specifically in the cytoplasm of basal acinar cells of the MGs, spontaneously develop DED/MGD. These mice display basal acinar cell hyperplasia, impaired meibocyte differentiation, reduced meibum production, and chronic inflammation. We found that the key mechanisms driving DED/MGD onset and progression-whose severity increases with age- include early NF-κB activation through both canonical and alternative pathways, upregulation of Cyclin D1, and downregulation of PPARγ. Transcriptomic analysis reveals in our murine model upregulation of inflammatory and immune-related genes, mirroring patterns observed in human DED and MGD, and, interestingly, it also uncover genetic changes characteristic of age-related DED and MGD, including the deregulation of genes associated with hormone and lipid metabolism, acinar cell proliferation and differentiation, and cell junctions. Overall, our data establish a direct link between increased cytoplasmic IKKα levels, impaired epithelial basal acinar cell differentiation, NF-κB activation, and the development of DED/MGD. Notably, elevated IKKα protein and mRNA levels are also detected in conjunctival tissues from DED patients, validating C-IKKα mice as a valuable model for investigating disease mechanisms, biomarkers, and therapeutic strategies. Biological sciences/Cell biology Health sciences/Pathogenesis/Inflammation/Chronic inflammation IKKα NF-κB Meibomian gland inflammation dry eye disease meibomian gland dysfunction aging Full Text Additional Declarations There is no duality of interest Tables are available in the Supplementary Files section. Supplementary Files Fulllengthuncroppedoriginalwesternblots.pdf Supplemental material: uncropped WBs Table1.xlsx Table 1 SuppleminformationGarciaGarciaVA.pdf Supplementary information Table2.xlsx Table 2 Cite Share Download PDF Status: Under Review 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-9300712","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":620160773,"identity":"ba549776-92e9-43f7-af19-2387ec6ab281","order_by":0,"name":"M. 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AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"IKKα, NF-κB, Meibomian gland, inflammation, dry eye disease, meibomian gland dysfunction, aging","lastPublishedDoi":"10.21203/rs.3.rs-9300712/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-9300712/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Dry eye disease (DED) is a multifactorial ocular surface disorder affecting over 20% of adults over 40, with higher prevalence in the elderly. Meibomian gland dysfuncton(MGD) is its leading cause. The meibomian glands (MGs) are holocrine sebaceous glands composed of acini, where basal cells differentiate into meibocytes that produce meibum, a lipid-rich substance essential for tear film stability and preventing evaporation. Inflammation is central to DED/MGD pathophysiology, driving a self-perpetuating cycle that disrupts innate and adaptive immunity and promotes chronicity. Overactivation of NF-κB -a key mediator of inflammation and aging- on the ocular surface likely plays a critical role in DED/MGD, though the mechanisms of its activation remain unclear, potentially limiting treatment efficacy.\r\nHere, we demonstrate that C-IKKα transgenic mice, which express exogenous IKKα specifically in the cytoplasm of basal acinar cells of the MGs, spontaneously develop DED/MGD. These mice display basal acinar cell hyperplasia, impaired meibocyte differentiation, reduced meibum production, and chronic inflammation. We found that the key mechanisms driving DED/MGD onset and progression-whose severity increases with age- include early NF-κB activation through both canonical and alternative pathways, upregulation of Cyclin D1, and downregulation of PPARγ. Transcriptomic analysis reveals in our murine model upregulation of inflammatory and immune-related genes, mirroring patterns observed in human DED and MGD, and, interestingly, it also uncover genetic changes characteristic of age-related DED and MGD, including the deregulation of genes associated with hormone and lipid metabolism, acinar cell proliferation and differentiation, and cell junctions. Overall, our data establish a direct link between increased cytoplasmic IKKα levels, impaired epithelial basal acinar cell differentiation, NF-κB activation, and the development of DED/MGD. Notably, elevated IKKα protein and mRNA levels are also detected in conjunctival tissues from DED patients, validating C-IKKα mice as a valuable model for investigating disease mechanisms, biomarkers, and therapeutic strategies.","manuscriptTitle":"Pathogenic Role of Cytoplasmic IKKα in Meibomian Gland Dysfunction and Dry Eye Disease","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-04-24 18:22:33","doi":"10.21203/rs.3.rs-9300712/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"cell-death-and-disease","isNatureJournal":false,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"cddis","sideBox":"Learn more about [Cell Death \u0026 Disease](http://www.nature.com/cddis/)","snPcode":"41419","submissionUrl":"https://mts-cddis.nature.com/cgi-bin/main.plex","title":"Cell Death \u0026 Disease","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Nature AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"bf78fba3-4c29-4c81-b427-48288e455c96","owner":[],"postedDate":"April 24th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":65994429,"name":"Biological sciences/Cell biology"},{"id":65994430,"name":"Health sciences/Pathogenesis/Inflammation/Chronic inflammation"}],"tags":[],"updatedAt":"2026-04-24T18:22:33+00:00","versionOfRecord":[],"versionCreatedAt":"2026-04-24 18:22:33","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-9300712","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-9300712","identity":"rs-9300712","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}