Extracellular vesicles deliver functional extrachromosomal DNA in FGFR2-amplified cancer of unknown primary

Extracellular vesicles deliver functional extrachromosomal DNA in FGFR2-amplified cancer of unknown primary | 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 Extracellular vesicles deliver functional extrachromosomal DNA in FGFR2-amplified cancer of unknown primary Irene Salamon, Giulia Gallerani, Jens Luebeck, Gianluca Storci, and 20 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6089646/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 Cancer cells actively release extracellular vesicles (EVs) into the tumor microenvironment, where they interact with both malignant and non-malignant cells, activating signaling pathways and reshaping the microenvironment. In this study, we investigated EVs secreted by FGFR2 -amplified cancers of unknown primary (CUPs), which generate extrachromosomal circular DNA (ecDNA) as a mechanism of oncogene amplification. We found that FGFR2 -containing ecDNA is packaged into both small and large EVs, horizontally transferred to recipient cells, and remains functionally active. Upon exposure to CUP-derived EVs—either by direct administration or co-culture—cancer (NCI-N87, THP1) and non-cancer (HUVEC, fibroblasts) cells internalized FGFR2 ecDNA, which was subsequently transcribed and translated to some extent. Functionally, CUP-derived EVs polarized THP1 cells toward an M2-like phenotype and promoted HUVEC proliferation. In vivo , xenografts generated from CUP cell lines released circulating FGFR2 + EVs, which mediated the systemic transfer of FGFR2 ecDNA to distant organs. Collectively, these findings demonstrate that tumor-derived EVs can propagate and horizontally transfer oncogenic ecDNA both in vitro and in vivo , providing a possible mechanistic basis for the high metastatic potential of this tumor type. CUP EVs ecDNA horizontal transfer Full Text Additional Declarations The authors declare no competing interests. Supplementary Files JEVSupplementaryMaterial.docx 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. 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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-6089646","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":421725839,"identity":"8ad00848-665c-44d0-a4be-48d6c046a830","order_by":0,"name":"Irene Salamon","email":"","orcid":"","institution":"IRCCS Azienda Ospedaliero-Universitaria di Bologna","correspondingAuthor":false,"prefix":"","firstName":"Irene","middleName":"","lastName":"Salamon","suffix":""},{"id":421725840,"identity":"98dc5b95-9e3a-4b8f-816a-6e8a2e6311b7","order_by":1,"name":"Giulia Gallerani","email":"","orcid":"","institution":"University of Bologna","correspondingAuthor":false,"prefix":"","firstName":"Giulia","middleName":"","lastName":"Gallerani","suffix":""},{"id":598754466,"identity":"0f84ec9d-34b5-4d2a-9b91-3c3794a019e9","order_by":2,"name":"Jens Luebeck","email":"","orcid":"","institution":"Department of Computer Science and Engineering, University of California, San Diego; 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In this study, we investigated EVs secreted by \u003cem\u003eFGFR2\u003c/em\u003e-amplified cancers of unknown primary (CUPs), which generate extrachromosomal circular DNA (ecDNA) as a mechanism of oncogene amplification. We found that \u003cem\u003eFGFR2\u003c/em\u003e-containing ecDNA is packaged into both small and large EVs, horizontally transferred to recipient cells, and remains functionally active. Upon exposure to CUP-derived EVs—either by direct administration or co-culture—cancer (NCI-N87, THP1) and non-cancer (HUVEC, fibroblasts) cells internalized \u003cem\u003eFGFR2\u003c/em\u003e ecDNA, which was subsequently transcribed and translated to some extent. Functionally, CUP-derived EVs polarized THP1 cells toward an M2-like phenotype and promoted HUVEC proliferation. \u003cem\u003eIn vivo\u003c/em\u003e, xenografts generated from CUP cell lines released circulating FGFR2\u003csup\u003e+\u003c/sup\u003e EVs, which mediated the systemic transfer of \u003cem\u003eFGFR2\u003c/em\u003e ecDNA to distant organs. 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