The Role of Progranulin in Glioblastoma Progression and Tumor Microenvironment Modulation

Progranulin Promotes Glioblastoma Malignancy and Immunosuppression via M2-TGF-β Axis: Therapeutic Efficacy of SORT1-Loaded HAMA Hydrogel Microspheres | 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 Progranulin Promotes Glioblastoma Malignancy and Immunosuppression via M2-TGF-β Axis: Therapeutic Efficacy of SORT1-Loaded HAMA Hydrogel Microspheres He Zhang, Shuai Yuan, Yawen Pan This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7403403/v2 This work is licensed under a CC BY 4.0 License Status: Posted Version 2 posted You are reading this latest preprint version Show more versions Abstract Glioblastoma multiforme (GBM) is the most aggressive primary brain tumor, characterized by rapid progression, therapeutic resistance, and a complex immunosuppressive tumor microenvironment (TME). Progranulin (GRN), a secreted glycoprotein implicated in various cancers, plays an underexplored role in GBM. This study investigates GRN's contributions to GBM malignancy and TME modulation, aiming to identify novel therapeutic targets. Integrated bioinformatics analyses of GEO (GSE90598) and TCGA datasets revealed significant GRN upregulation in GBM tissues, correlating with poor overall and progression-free survival (p 1). GRN expression showed strong positive associations with M2 macrophage marker CD163 at both protein (IHC, R = 0.56, p < 0.001) and mRNA levels (TCGA, R = 0.53, p < 0.001). CIBERSORT and ESTIMATE analyses indicated elevated M2 macrophage infiltration and higher immune/stromal scores in high-GRN groups, underscoring GRN's role in fostering immunosuppression. In vitro, GRN knockdown (siGRN) in GBM cell lines (A172) significantly suppressed proliferation (CCK-8), migration (scratch assay), invasion (Transwell), and clonogenicity (p < 0.05). Coculture with macrophages demonstrated that siGRN shifted polarization from M2 (CD206⁺) to M1 (CD86⁺), reducing immunosuppressive cytokines IL-10 and TGF-β1. Transwell assays confirmed GRN's macrophage-dependent promotion of GBM invasion, abrogated by TGF-β receptor inhibitor LY3200882, highlighting the GRN-TGF-β-macrophage axis. In vivo, SORT1-loaded hyaluronic acid methacryloyl (HAMA) hydrogel microspheres, fabricated via microfluidics (70-90 μm diameter, 6-day degradation), were intratumorally injected into nude mouse GBM xenografts. This significantly inhibited tumor growth compared to controls (p < 0.05), demonstrating GRN-targeted therapy's potential. Collectively, GRN drives GBM aggressiveness and TME immunosuppression via M2 polarization and TGF-β signaling. Utilizing SORT1-loaded HAMA microspheres to target and clear GRN proteins provides a promising postoperative treatment strategy for inhibiting GBM recurrence and improving prognosis. Glioblastoma Progranulin Tumor Microenvironment M2 Macrophage Polarization TGF-β Signaling Immunosuppression Full Text Additional Declarations The authors declare no competing interests. Cite Share Download PDF Status: Posted Version 2 posted You are reading this latest preprint version Show more versions 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-7403403","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":521231622,"identity":"0c8caefb-46bf-4e15-aee0-fb60b7920234","order_by":0,"name":"He Zhang","email":"","orcid":"","institution":"Lanzhou University Second Hospital","correspondingAuthor":false,"prefix":"","firstName":"He","middleName":"","lastName":"Zhang","suffix":""},{"id":521231623,"identity":"d8811795-1241-43aa-a074-fed13ef57fca","order_by":1,"name":"Shuai Yuan","email":"","orcid":"","institution":"Lanzhou University Second 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Microspheres\u003c/strong\u003e\u003c/p\u003e","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":"Glioblastoma, Progranulin, Tumor Microenvironment, M2 Macrophage Polarization, TGF-β Signaling, Immunosuppression","lastPublishedDoi":"10.21203/rs.3.rs-7403403/v2","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7403403/v2","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eGlioblastoma multiforme (GBM) is the most aggressive primary brain tumor, characterized by rapid progression, therapeutic resistance, and a complex immunosuppressive tumor microenvironment (TME). Progranulin (GRN), a secreted glycoprotein implicated in various cancers, plays an underexplored role in GBM. This study investigates GRN's contributions to GBM malignancy and TME modulation, aiming to identify novel therapeutic targets. Integrated bioinformatics analyses of GEO (GSE90598) and TCGA datasets revealed significant GRN upregulation in GBM tissues, correlating with poor overall and progression-free survival (p \u0026lt; 0.05, HR \u0026gt; 1). GRN expression showed strong positive associations with M2 macrophage marker CD163 at both protein (IHC, R = 0.56, p \u0026lt; 0.001) and mRNA levels (TCGA, R = 0.53, p \u0026lt; 0.001). CIBERSORT and ESTIMATE analyses indicated elevated M2 macrophage infiltration and higher immune/stromal scores in high-GRN groups, underscoring GRN's role in fostering immunosuppression. In vitro, GRN knockdown (siGRN) in GBM cell lines (A172) significantly suppressed proliferation (CCK-8), migration (scratch assay), invasion (Transwell), and clonogenicity (p \u0026lt; 0.05). Coculture with macrophages demonstrated that siGRN shifted polarization from M2 (CD206⁺) to M1 (CD86⁺), reducing immunosuppressive cytokines IL-10 and TGF-β1. Transwell assays confirmed GRN's macrophage-dependent promotion of GBM invasion, abrogated by TGF-β receptor inhibitor LY3200882, highlighting the GRN-TGF-β-macrophage axis. In vivo, SORT1-loaded hyaluronic acid methacryloyl (HAMA) hydrogel microspheres, fabricated via microfluidics (70-90 μm diameter, 6-day degradation), were intratumorally injected into nude mouse GBM xenografts. This significantly inhibited tumor growth compared to controls (p \u0026lt; 0.05), demonstrating GRN-targeted therapy's potential. Collectively, GRN drives GBM aggressiveness and TME immunosuppression via M2 polarization and TGF-β signaling. Utilizing SORT1-loaded HAMA microspheres to target and clear GRN proteins provides a promising postoperative treatment strategy for inhibiting GBM recurrence and improving prognosis.\u003c/p\u003e","manuscriptTitle":"Progranulin Promotes Glioblastoma Malignancy and Immunosuppression via M2-TGF-β Axis: Therapeutic Efficacy of SORT1-Loaded HAMA Hydrogel Microspheres","msid":"","msnumber":"","nonDraftVersions":[{"code":2,"date":"2025-12-10 21:42:12","doi":"10.21203/rs.3.rs-7403403/v2","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}},{"code":1,"date":"2025-09-26 16:09:43","doi":"10.21203/rs.3.rs-7403403/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":"8e4ddc41-af0a-447d-ac38-74e884f0b300","owner":[],"postedDate":"December 10th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-11-06T06:38:33+00:00","versionOfRecord":[],"versionCreatedAt":"2025-12-10 21:42:12","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v2","identity":"rs-7403403","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7403403","identity":"rs-7403403","version":["v2"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}