Engineered CXCR3-A expression enhances the trafficking and efficacy of intracerebroventricularly delivered B7-H3-targeting CAR T cells against diffuse intrinsic pontine glioma

Engineered CXCR3-A expression enhances the trafficking and efficacy of intracerebroventricularly delivered B7-H3-targeting CAR T cells against diffuse intrinsic pontine glioma | 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 Engineered CXCR3-A expression enhances the trafficking and efficacy of intracerebroventricularly delivered B7-H3-targeting CAR T cells against diffuse intrinsic pontine glioma Nicholas Vitanza, Edward Song, Andrea Timpanaro, Michael Meechan, and 13 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6272325/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 11 Nov, 2025 Read the published version in Nature Communications → Version 1 posted You are reading this latest preprint version Abstract Diffuse intrinsic pontine glioma (DIPG) is a fatal brainstem tumor desperately in need of better treatments. CAR T cell therapies for DIPG have demonstrated clinical tolerability and bioactivity, but not universal benefit. A major obstacle is insufficient CAR T cell trafficking to the tumor. As our clinical trials have demonstrated locoregional elevation of CXCL10, a ligand of the chemokine receptor CXCR3, we aimed to leverage this CXCL10 to enhance cell trafficking by engineering our B7-H3-targeting CAR T cells to overexpress CXCR3 variants. Here, we demonstrate that, compared to unmodified B7-H3 CAR T cells, CXCR3-A-modified CAR T cells migrate more efficiently toward CXCR3 ligands in vitro, and when delivered intracerebroventricularly in orthotopic DIPG mouse models, CXCR3-A-modified CAR T cells show enhanced trafficking into the tumor and improved therapeutic efficacy. Overall, our data support the potential for engineering CXCR3-A expression to enhance CAR T cell trafficking and efficacy against DIPG. Biological sciences/Cancer/CNS cancer Biological sciences/Cancer/Paediatric cancer Biological sciences/Cancer/Cancer therapy/Cancer immunotherapy Biological sciences/Immunology/Chemokines Biological sciences/Cell biology/Cell migration/Chemotaxis Full Text Additional Declarations Yes there is potential Competing Interest. E.Z.S., J.B.F., M.C.J., and N.A.V. are inventors on issued and pending patents related to CAR T cell therapies. M.C.J. holds equity in and is the Chief Scientific Officer of BrainChild Bio, Inc. M.C.J. holds equity in, is a Board Observer for and serves as a member of the Joint Steering Committee of Umoja Biopharma, Inc. N.A.V. holds equity in and serves as the Scientific Advisory Board Chair for BrainChild Bio, Inc. All other authors declare no competing interests. Cite Share Download PDF Status: Published Journal Publication published 11 Nov, 2025 Read the published version in Nature Communications → 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-6272325","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":435481056,"identity":"4bfdd11b-845b-4301-8ddd-7c99d00baeb1","order_by":0,"name":"Nicholas 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there is potential Competing Interest.\nE.Z.S., J.B.F., M.C.J., and N.A.V. are inventors on issued and pending patents related to CAR T cell therapies. M.C.J. holds equity in and is the Chief Scientific Officer of BrainChild Bio, Inc. M.C.J. holds equity in, is a Board Observer for and serves as a member of the Joint Steering Committee of Umoja Biopharma, Inc. N.A.V. holds equity in and serves as the Scientific Advisory Board Chair for BrainChild Bio, Inc. 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CAR T cell therapies for DIPG have demonstrated clinical tolerability and bioactivity, but not universal benefit. A major obstacle is insufficient CAR T cell trafficking to the tumor. As our clinical trials have demonstrated locoregional elevation of CXCL10, a ligand of the chemokine receptor CXCR3, we aimed to leverage this CXCL10 to enhance cell trafficking by engineering our B7-H3-targeting CAR T cells to overexpress CXCR3 variants. Here, we demonstrate that, compared to unmodified B7-H3 CAR T cells, CXCR3-A-modified CAR T cells migrate more efficiently toward CXCR3 ligands in vitro, and when delivered intracerebroventricularly in orthotopic DIPG mouse models, CXCR3-A-modified CAR T cells show enhanced trafficking into the tumor and improved therapeutic efficacy. 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