Activation of Wnt/β-catenin signaling in histologically non-dysplastic Proliferative Verrucous Leukoplakia

Activation of Wnt/β-catenin signaling in histologically non-dysplastic Proliferative Verrucous Leukoplakia | 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 Activation of Wnt/β-catenin signaling in histologically non-dysplastic Proliferative Verrucous Leukoplakia Daniel Peña-Oyarzún, Andrea Maturana-Ramírez, Montserrat Reyes This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8846248/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 01 May, 2026 Read the published version in BMC Oral Health → Version 1 posted 10 You are reading this latest preprint version Abstract Purpose: Oral potentially malignant disorders (OPMD) are lesions that precede oral cancer in approximately 15-40% of cases. Leukoplakia is the most frequent type of OPMD, and according to the roughness of the lesion surface is clinically classified as homogeneous oral leukoplakia (HOL) or proliferative verrucous leukoplakia (PVL). Compared to HOL, PVL is less frequent and is associated with higher malignant transformation rates. This difference is even more evident in lesions histopathologically diagnosed as non-dysplastic, including hyperplasia and hyperkeratosis. To the date, there is no signaling pathway that may explain this. We and others have shown that the Wnt/β-catenin pathway, characterized by stabilization and nuclear translocation of β-catenin for expression proliferation and survival genes, is key for dysplastic HOL carcinogenesis. The role of the Wnt/β-catenin pathway in dysplastic and non-dysplastic PVL remains unknown. Methods: 53 retrospective biopsies from patients with PVL and HOL were used. Histological diagnosis was performed with Hematoxylin & Eosin (H&E) staining, while Wnt/β-catenin activity of signaling pathway was evaluated by immunohistochemical expression of the ligand Wnt3a and nuclear localization of β-catenin, as well as downstream proliferation markers Cyclin D1 and Ki-67. Results: PVL lesions histologically diagnosed as hyperplasia/hyperkeratosis (non-dysplastic), displayed significantly higher expression of Wnt3a and nuclear β-catenin, as well as Cyclin D1 and Ki-67, compared to non-dysplastic HOL and healthy oral mucosa. No significant differences were observed when comparing Wnt3a, nuclear β-catenin, Cyclin D1 and Ki-67 expression between non-dysplastic HOL and healthy oral mucosa. Both dysplastic PVL and HOL samples, compared to healthy oral mucosa, presented significant upregulation of Wnt3a, nuclear β-catenin, Cyclin D1 and Ki-67. No significant differences in Wnt3a, nuclear β-catenin, Cyclin D1 and Ki-67 expression were observed between dysplastic PVL and HOL. Conclusions: We conclude that, in clear contrast to non-dysplastic HOL, Wnt/β-catenin activity is increased in non-dysplastic PVL, which correlates with the expression of downstream malignant proliferation markers. Additionally, both dysplastic PVL and HOL present high Wnt/β-catenin activity, regardless of the dysplasia grade. Proliferative verrucous leukoplakia homogeneous leukoplakia dysplasia Wnt/β-catenin signaling pathway Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 01 May, 2026 Read the published version in BMC Oral Health → Version 1 posted Editorial decision: Revision requested 30 Mar, 2026 Reviews received at journal 24 Mar, 2026 Reviews received at journal 23 Mar, 2026 Reviewers agreed at journal 23 Mar, 2026 Reviewers agreed at journal 17 Mar, 2026 Reviewers invited by journal 26 Feb, 2026 Editor assigned by journal 26 Feb, 2026 Editor invited by journal 18 Feb, 2026 Submission checks completed at journal 16 Feb, 2026 First submitted to journal 16 Feb, 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-8846248","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":599268587,"identity":"986231f7-7d7d-4a59-96ce-c0c2f6be44c0","order_by":0,"name":"Daniel Peña-Oyarzún","email":"","orcid":"","institution":"San Sebastián University","correspondingAuthor":false,"prefix":"","firstName":"Daniel","middleName":"","lastName":"Peña-Oyarzún","suffix":""},{"id":599268588,"identity":"63929a20-e5bb-45e5-ba6d-e425ed2a53d0","order_by":1,"name":"Andrea Maturana-Ramírez","email":"","orcid":"","institution":"University of Chile","correspondingAuthor":false,"prefix":"","firstName":"Andrea","middleName":"","lastName":"Maturana-Ramírez","suffix":""},{"id":599268589,"identity":"103c5085-b395-449e-b3ef-afcc339194f4","order_by":2,"name":"Montserrat Reyes","email":"data:image/png;base64,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","orcid":"","institution":"University of Chile","correspondingAuthor":true,"prefix":"","firstName":"Montserrat","middleName":"","lastName":"Reyes","suffix":""}],"badges":[],"createdAt":"2026-02-11 02:23:45","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8846248/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8846248/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1186/s12903-026-08361-z","type":"published","date":"2026-05-01T15:57:22+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":108438056,"identity":"42c3fe53-6dc3-4391-9500-f06682b86001","added_by":"auto","created_at":"2026-05-04 16:06:21","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1225782,"visible":true,"origin":"","legend":"","description":"","filename":"Manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8846248/v1_covered_3063825c-33a3-4ed7-b7ac-22b4c99a6f1d.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Activation of Wnt/β-catenin signaling in histologically non-dysplastic Proliferative Verrucous Leukoplakia","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"bmc-oral-health","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"ohea","sideBox":"Learn more about [BMC Oral Health](http://bmcoralhealth.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/ohea/default.aspx","title":"BMC Oral Health","twitterHandle":"BMC_series","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Proliferative verrucous leukoplakia, homogeneous leukoplakia, dysplasia, Wnt/β-catenin signaling pathway","lastPublishedDoi":"10.21203/rs.3.rs-8846248/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8846248/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003ePurpose:\u003c/strong\u003e Oral potentially malignant disorders (OPMD) are lesions that precede oral cancer in approximately 15-40% of cases. Leukoplakia is the most frequent type of OPMD, and according to the roughness of the lesion surface is clinically classified as homogeneous oral leukoplakia (HOL) or proliferative verrucous leukoplakia (PVL). Compared to HOL, PVL is less frequent and is associated with higher malignant transformation rates. This difference is even more evident in lesions histopathologically diagnosed as non-dysplastic, including hyperplasia and hyperkeratosis. To the date, there is no signaling pathway that may explain this. We and others have shown that the Wnt/β-catenin pathway, characterized by stabilization and nuclear translocation of β-catenin for expression proliferation and survival genes, is key for dysplastic HOL carcinogenesis. The role of the Wnt/β-catenin pathway in dysplastic and non-dysplastic PVL remains unknown.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods:\u003c/strong\u003e 53 retrospective biopsies from patients with PVL and HOL were used. Histological diagnosis was performed with Hematoxylin \u0026amp; Eosin (H\u0026amp;E) staining, while Wnt/β-catenin activity of signaling pathway was evaluated by immunohistochemical expression of the ligand Wnt3a and nuclear localization of β-catenin, as well as downstream proliferation markers Cyclin D1 and Ki-67.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults:\u003c/strong\u003e PVL lesions histologically diagnosed as hyperplasia/hyperkeratosis (non-dysplastic), displayed significantly higher expression of Wnt3a and nuclear β-catenin, as well as Cyclin D1 and Ki-67, compared to non-dysplastic HOL and healthy oral mucosa. No significant differences were observed when comparing Wnt3a, nuclear β-catenin, Cyclin D1 and Ki-67 expression between non-dysplastic HOL and healthy oral mucosa. Both dysplastic PVL and HOL samples, compared to healthy oral mucosa, presented significant upregulation of Wnt3a, nuclear β-catenin, Cyclin D1 and Ki-67. No significant differences in Wnt3a, nuclear β-catenin, Cyclin D1 and Ki-67 expression were observed between dysplastic PVL and HOL.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusions:\u003c/strong\u003e We conclude that, in clear contrast to non-dysplastic HOL, Wnt/β-catenin activity is increased in non-dysplastic PVL, which correlates with the expression of downstream malignant proliferation markers. Additionally, both dysplastic PVL and HOL present high Wnt/β-catenin activity, regardless of the dysplasia grade.\u003c/p\u003e","manuscriptTitle":"Activation of Wnt/β-catenin signaling in histologically non-dysplastic Proliferative Verrucous Leukoplakia","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-03-03 19:59:06","doi":"10.21203/rs.3.rs-8846248/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-03-30T16:09:44+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-03-24T15:27:36+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-03-23T17:39:38+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"262762204590638565762562820919408624285","date":"2026-03-23T14:38:54+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"283856816166412983936599400834386198237","date":"2026-03-17T15:08:41+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-02-26T16:06:32+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-02-26T15:51:04+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2026-02-18T05:05:45+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-02-16T22:12:56+00:00","index":"","fulltext":""},{"type":"submitted","content":"BMC Oral Health","date":"2026-02-16T22:08:05+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"bmc-oral-health","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"ohea","sideBox":"Learn more about [BMC Oral Health](http://bmcoralhealth.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/ohea/default.aspx","title":"BMC Oral Health","twitterHandle":"BMC_series","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"0543afd9-e6c5-4e59-8b3e-34c49ce1e2f2","owner":[],"postedDate":"March 3rd, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2026-05-04T16:06:17+00:00","versionOfRecord":{"articleIdentity":"rs-8846248","link":"https://doi.org/10.1186/s12903-026-08361-z","journal":{"identity":"bmc-oral-health","isVorOnly":false,"title":"BMC Oral Health"},"publishedOn":"2026-05-01 15:57:22","publishedOnDateReadable":"May 1st, 2026"},"versionCreatedAt":"2026-03-03 19:59:06","video":"","vorDoi":"10.1186/s12903-026-08361-z","vorDoiUrl":"https://doi.org/10.1186/s12903-026-08361-z","workflowStages":[]},"version":"v1","identity":"rs-8846248","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8846248","identity":"rs-8846248","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}