Anti-VEGF vascular remodeling drives germinal center B cell-rich tertiary lymphoid structures during antibody-toxin and anti-CD40 combination therapy in glioblastoma

Anti-VEGF vascular remodeling drives germinal center B cell-rich tertiary lymphoid structures during antibody-toxin and anti-CD40 combination therapy in glioblastoma | 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 Anti-VEGF vascular remodeling drives germinal center B cell-rich tertiary lymphoid structures during antibody-toxin and anti-CD40 combination therapy in glioblastoma Anjali Barnwal, Weixuan Yan, Sarah Cook, Thomas Urup, Kevin Stevenson, and 11 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9350019/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 Glioblastoma (GBM) is characterized by aberrant neovascularization and angiogenesis. Bevacizumab (Avastin), a monoclonal antibody targeting vascular endothelial growth factor (VEGF), is clinically used with chemotherapy for patients with GBM, yet its impact on the tumor immune microenvironment remains poorly understood. Transcriptomic profiling of matched pre- and post-treatment tumors from patients with recurrent GBM treated with bevacizumab plus the topoisomerase inhibitor irinotecan revealed significant immune reprogramming in responders, with post-treatment tumors displaying features indicative of tertiary lymphoid structure (TLS) formation and enhanced antitumor immunity. In preclinical orthotopic glioma models, combining αVEGF therapy with intratumoral epidermal growth factor receptor (EGFR)-targeted cytotoxic therapy and αCD40 immunotherapy normalized the tumor vasculature, promoted lymphatic vessel growth, induced tumor cell killing, increased intratumoral T cells, plasma cells, germinal center B cells, and antigen-presenting and tissue-repair-associated macrophages, while reducing immunosuppression and supporting mature TLS formation with an antitumor immune phenotype. Functionally, the tumor-, immune-, and vascular-targeted combination therapy significantly improved tumor control, extended survival, and induced durable antitumor memory in glioma models. Collectively, these results identify VEGF as a central regulator of the vascular-immune axis during cytotoxic EGFR+αCD40 therapy in GBM and provide a strong rationale for combining vascular-, tumor-, and immune-targeted approaches to overcome therapeutic resistance and improve outcomes for patients with GBM. Biological sciences/Cancer/CNS cancer Health sciences/Oncology/Cancer/Cancer therapy/Cancer immunotherapy Full Text Additional Declarations Yes there is potential Competing Interest. V.C. and D.D.B. are inventors of ATC patent applications. A.D. serves as an advisory board member for Orbus Therapeutics, Telix Therapeutics, and Altido, and receives clinical research support (to the institution) from Orbus Therapeutics, Exvade Bioscience, Creosalus, and Biodexa. D.M.A. serves as an advisory board member for Immunogenesis, MAIA Biotechnology, and Diverse Biotech and as a consultant for the Jackson Laboratory. The other authors declare that they have no competing interests. Supplementary Files SupplementaryFigure1Final.tif Supplementary Figure 1 SupplementaryFigure2Final.tif Supplementary Figure 2 SupplementaryFigure3Final.tif Supplementary Figure 3 SupplementaryFigure4Final.tif Supplementary Figure 4 SupplementaryFigure5Final.tif Supplementary Figure 5 SupplementaryFigure6Final.tif Supplementary Figure 6 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-9350019","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":624420244,"identity":"c3c5341c-030c-4648-91f3-e5b51822e7ae","order_by":0,"name":"Anjali Barnwal","email":"","orcid":"https://orcid.org/0000-0001-7190-8444","institution":"Duke University Medical Center","correspondingAuthor":false,"prefix":"","firstName":"Anjali","middleName":"","lastName":"Barnwal","suffix":""},{"id":624420245,"identity":"30ba1199-c598-44c3-8468-355117cfa442","order_by":1,"name":"Weixuan Yan","email":"","orcid":"","institution":"Duke 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A.D. serves as an advisory board member for Orbus Therapeutics, Telix Therapeutics, and Altido, and receives clinical research support (to the institution) from Orbus Therapeutics, Exvade Bioscience, Creosalus, and Biodexa. D.M.A. serves as an advisory board member for Immunogenesis, MAIA Biotechnology, and Diverse Biotech and as a consultant for the Jackson Laboratory. \r\nThe other authors declare that they have no competing interests.","formattedTitle":"Anti-VEGF vascular remodeling drives germinal center B cell-rich tertiary lymphoid structures during antibody-toxin and anti-CD40 combination therapy in glioblastoma","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":false,"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":"nature-portfolio","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"","title":"Nature Portfolio","twitterHandle":"","acdcEnabled":false,"dfaEnabled":false,"editorialSystem":"ejp","reportingPortfolio":"","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-9350019/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-9350019/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Glioblastoma (GBM) is characterized by aberrant neovascularization and angiogenesis. Bevacizumab (Avastin), a monoclonal antibody targeting vascular endothelial growth factor (VEGF), is clinically used with chemotherapy for patients with GBM, yet its impact on the tumor immune microenvironment remains poorly understood. Transcriptomic profiling of matched pre- and post-treatment tumors from patients with recurrent GBM treated with bevacizumab plus the topoisomerase inhibitor irinotecan revealed significant immune reprogramming in responders, with post-treatment tumors displaying features indicative of tertiary lymphoid structure (TLS) formation and enhanced antitumor immunity. 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