Multi-omic characterization of MAP4 in pancreatic adenocarcinoma progression: epitranscriptomic networks, EMT Activation, and stromal-immune dynamics

Multi-omic characterization of MAP4 in pancreatic adenocarcinoma progression: epitranscriptomic networks, EMT Activation, and stromal-immune dynamics | 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 Multi-omic characterization of MAP4 in pancreatic adenocarcinoma progression: epitranscriptomic networks, EMT Activation, and stromal-immune dynamics Longyan Zuo, Yunliang Yu, Aining Ju, Xinmiao Xian, Yingfei Liang, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6702603/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 Background Pancreatic adenocarcinoma (PAAD), particularly pancreatic ductal adenocarcinoma (PDAC), is a lethal malignancy with limited therapeutic advancements. Microtubule-associated protein 4 (MAP4), implicated in tumor progression in other cancers, has not been systematically explored in PAAD. This study aims to investigate the role of MAP4 in the progression of PAAD, its association with tumor stemness, signaling pathways, and the tumor microenvironment (TME). Methods MAP4 expression was analyzed in 80 clinical specimens (60 PAAD and 20 normal tissues) using immunofluorescence. Bioinformatics analyses of 179 TCGA-PAAD samples were conducted to assess the prognostic significance of MAP4, its relationship with tumor heterogeneity, stemness, and immune profiles. Functional studies included silencing MAP4 in PAAD cell lines to evaluate effects on proliferation, migration, invasion, and signaling pathway activation, particularly TGF-β and epithelial-mesenchymal transition (EMT) pathways. Results MAP4 was significantly upregulated in PAAD tissues, correlating with poor differentiation ( P = 0.031) and shorter disease-free ( P = 4.0e − 4) and progression-free intervals ( P = 8.5e − 3). MAP4 silencing suppressed proliferation, migration, and invasion ( P 0.7), EMT markers (MMP2: r = 0.61), and RNA modification pathways (m 6 A/m 5 C/m 1 A regulators, P < 0.05). Negative associations with mRNA stemness indices (mRNAsi: r = − 0.48) suggested stemness modulation via epitranscriptomic networks. TME analysis revealed MAP4’s stromal dominance, with strong cancer-associated fibroblasts (CAFs) correlations (PDGFRB: r = 0.71; FAP: r = 0.68) and immunosuppressive features (PD-L1: r = 0.59; TIDE scores: P = 0.016). Single-cell data localized MAP4 to fibroblasts and endothelial cells, supporting stromal crosstalk. Conclusions MAP4 drives the progression of PAAD through multiple mechanisms, including the enhancement of TGF-β/EMT signaling, modulation of RNA modification networks to reduce stemness, and promotion of an immunosuppressive TME through CAFs and upregulation of immune checkpoints. These findings highlight the potential of MAP4 as both a prognostic biomarker and a therapeutic target, suggesting opportunities for combined strategies that address both stromal and immune components in treating PAAD. Microtubule-associated protein 4 Pancreatic adenocarcinoma Prognosis Tumor stemness Tumor heterogeneity Immune infiltration Full Text Additional Declarations No competing interests reported. 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. 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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-6702603","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":459312225,"identity":"34b78dc0-6641-4ca5-b873-04347d3286eb","order_by":0,"name":"Longyan Zuo","email":"","orcid":"","institution":"Liaocheng People’s Hospital","correspondingAuthor":false,"prefix":"","firstName":"Longyan","middleName":"","lastName":"Zuo","suffix":""},{"id":459312226,"identity":"46a2e257-b5b9-401d-9f77-47b331e03cac","order_by":1,"name":"Yunliang Yu","email":"","orcid":"","institution":"Yantai Affiliated Hospital of Binzhou Medical 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01:38:12","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6702603/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6702603/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":85621722,"identity":"a0c16183-9df6-4f75-9337-363953c207e6","added_by":"auto","created_at":"2025-06-29 16:16:40","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2069132,"visible":true,"origin":"","legend":"","description":"","filename":"Manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6702603/v1_covered_a5881057-960c-4567-90f2-4a64cef7f6c0.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Multi-omic characterization of MAP4 in pancreatic adenocarcinoma progression: epitranscriptomic networks, EMT Activation, and stromal-immune dynamics","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":"Microtubule-associated protein 4, Pancreatic adenocarcinoma, Prognosis, Tumor stemness, Tumor heterogeneity, Immune infiltration","lastPublishedDoi":"10.21203/rs.3.rs-6702603/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6702603/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e \u003cp\u003ePancreatic adenocarcinoma (PAAD), particularly pancreatic ductal adenocarcinoma (PDAC), is a lethal malignancy with limited therapeutic advancements. Microtubule-associated protein 4 (MAP4), implicated in tumor progression in other cancers, has not been systematically explored in PAAD. This study aims to investigate the role of MAP4 in the progression of PAAD, its association with tumor stemness, signaling pathways, and the tumor microenvironment (TME).\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e \u003cp\u003eMAP4 expression was analyzed in 80 clinical specimens (60 PAAD and 20 normal tissues) using immunofluorescence. Bioinformatics analyses of 179 TCGA-PAAD samples were conducted to assess the prognostic significance of MAP4, its relationship with tumor heterogeneity, stemness, and immune profiles. Functional studies included silencing MAP4 in PAAD cell lines to evaluate effects on proliferation, migration, invasion, and signaling pathway activation, particularly TGF-β and epithelial-mesenchymal transition (EMT) pathways.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eMAP4 was significantly upregulated in PAAD tissues, correlating with poor differentiation (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.031) and shorter disease-free (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;4.0e\u0026thinsp;\u0026minus;\u0026thinsp;4) and progression-free intervals (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.5e\u0026thinsp;\u0026minus;\u0026thinsp;3). MAP4 silencing suppressed proliferation, migration, and invasion (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). MAP4 strongly correlated with TGF-β signaling (\u003cem\u003er\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.7), EMT markers (MMP2: \u003cem\u003er\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.61), and RNA modification pathways (m\u003csup\u003e6\u003c/sup\u003eA/m\u003csup\u003e5\u003c/sup\u003eC/m\u003csup\u003e1\u003c/sup\u003eA regulators, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Negative associations with mRNA stemness indices (mRNAsi: \u003cem\u003er\u003c/em\u003e\u0026thinsp;=\u0026thinsp;\u0026minus;\u0026thinsp;0.48) suggested stemness modulation via epitranscriptomic networks. TME analysis revealed MAP4\u0026rsquo;s stromal dominance, with strong cancer-associated fibroblasts (CAFs) correlations (PDGFRB: \u003cem\u003er\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.71; FAP: \u003cem\u003er\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.68) and immunosuppressive features (PD-L1: \u003cem\u003er\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.59; TIDE scores: \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.016). Single-cell data localized MAP4 to fibroblasts and endothelial cells, supporting stromal crosstalk.\u003c/p\u003e\u003ch2\u003eConclusions\u003c/h2\u003e \u003cp\u003eMAP4 drives the progression of PAAD through multiple mechanisms, including the enhancement of TGF-β/EMT signaling, modulation of RNA modification networks to reduce stemness, and promotion of an immunosuppressive TME through CAFs and upregulation of immune checkpoints. These findings highlight the potential of MAP4 as both a prognostic biomarker and a therapeutic target, suggesting opportunities for combined strategies that address both stromal and immune components in treating PAAD.\u003c/p\u003e","manuscriptTitle":"Multi-omic characterization of MAP4 in pancreatic adenocarcinoma progression: epitranscriptomic networks, EMT Activation, and stromal-immune dynamics","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-05-21 15:44:32","doi":"10.21203/rs.3.rs-6702603/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":"c86bad9b-c8d8-4075-9b1c-8adbd9dc7903","owner":[],"postedDate":"May 21st, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-08-07T17:53:18+00:00","versionOfRecord":[],"versionCreatedAt":"2025-05-21 15:44:32","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6702603","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6702603","identity":"rs-6702603","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}