Tetraspanin-10 as a Potential Marker for Disease-Associated Extracellular Vesicles

Tetraspanin-10 as a Potential Marker for Disease-Associated Extracellular Vesicles | 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 Tetraspanin-10 as a Potential Marker for Disease-Associated Extracellular Vesicles Makoto Sumazaki This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7690699/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 This exploratory study introduces the concept of disease-associated Extracellular Vesicles (EVs), defined as EVs secreted under pathological conditions, and identifies a potential molecular marker for their characterization. However, disease-associated EVs constitute only a small fraction of circulating EVs, making their selective isolation and characterization a critical challenge. Identifying specific surface markers is essential for isolating these vesicles and advancing both fundamental EV biology research and clinical applications. It is valuable to analyze EVs that are derived directly from pathological tissues, rather than the heterogeneous EVs in the blood. A comprehensive proteome of tissue-exudative EVs (Te-EVs) obtained from the culture supernatant of freshly resected colorectal cancer tissues has been reported. We used this EV proteome dataset to identify candidates for novel EV surface marker specific to disease-associated EVs in cancer. We screened multi-pass membrane proteins that were suitable for immunoaffinity EV isolation and discovered that Tetraspanin-10 (TSPAN10) was specific to cancer Te-EVs. As a result of referring to RNA sequencing published by The Cancer Genome Atlas and FANTOM5, the physiological expression of TSPAN10 was localized to the retina, while almost all cases of solid tumor expressed TSPAN10 mRNA. In colorectal cancer, TSPAN10 mRNA expression was significantly associated with tumor stage and prognosis. We evaluated the diagnostic potential of TSPAN10 using serum from patients with colorectal cancer. TSPAN10 signals measured by enzyme-linked immunosorbent assay increased in a stage-dependent manner and demonstrated favorable diagnostic potential with 83% sensitivity and 84% specificity. Transmission electron microscopy showed the membrane localization of TSPAN10 on an EV derived from a patient with advanced colorectal cancer. TSPAN10-expressing EVs may represent a subset of disease-associated EVs, offering diagnostic utility and advancing biological insights into EVs in cancer and other pathological conditions. Oncology Disease-associated extracellular vesicles TSPAN10 Colorectal cancer Multi-pass membrane protein Full Text Additional Declarations The authors declare no competing interests. Supplementary Figures S1 and S2 are not available with this version. Fig. S1. Workflow for the isolation and strategic use of tissue-exudative extracellular vesicles (Te-EVs) derived from freshly resected colorectal cancer (CRC) and adjacent normal tissues. This schematic was newly prepared based on the experimental procedures described in the previous report (10), with additional annotations to highlight the strategic rationale used in this study. Fig. S2. Representative images of immunohistological staining of TSPAN10 in normal adjacent colon mucosa (A) and CRC tissue (B), 4 × 10. TSPAN10, Tetraspanin-10; CRC, colorectal cancer 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. 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-7690699","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":519189424,"identity":"9e9c3e6b-465a-4b6c-812d-703cdb763e59","order_by":0,"name":"Makoto 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Workflow for the isolation and strategic use of tissue-exudative extracellular vesicles (Te-EVs) derived from freshly resected colorectal cancer (CRC) and adjacent normal tissues. This schematic was newly prepared based on the experimental procedures described in the previous report (10), with additional annotations to highlight the strategic rationale used in this study.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eFig. S2. Representative images of immunohistological staining of TSPAN10 in normal adjacent colon mucosa (A) and CRC tissue (B), 4 × 10. 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","order_by":1},{"identity":"baed3428-0bd3-4864-bdf5-f49b5735d48d","identifier":"10.13039/100007392","name":"Japan Association for the Advancement of Medical Equipment","awardNumber":"JAAME-2025Seed009","order_by":2}],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"Toho University","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":"Disease-associated extracellular vesicles, TSPAN10, Colorectal cancer, Multi-pass membrane protein","lastPublishedDoi":"10.21203/rs.3.rs-7690699/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7690699/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThis exploratory study introduces the concept of disease-associated Extracellular Vesicles (EVs), defined as EVs secreted under pathological conditions, and identifies a potential molecular marker for their characterization. However, disease-associated EVs constitute only a small fraction of circulating EVs, making their selective isolation and characterization a critical challenge. Identifying specific surface markers is essential for isolating these vesicles and advancing both fundamental EV biology research and clinical applications.\u003c/p\u003e\u003cp\u003eIt is valuable to analyze EVs that are derived directly from pathological tissues, rather than the heterogeneous EVs in the blood. A comprehensive proteome of tissue-exudative EVs (Te-EVs) obtained from the culture supernatant of freshly resected colorectal cancer tissues has been reported. We used this EV proteome dataset to identify candidates for novel EV surface marker specific to disease-associated EVs in cancer. We screened multi-pass membrane proteins that were suitable for immunoaffinity EV isolation and discovered that Tetraspanin-10 (TSPAN10) was specific to cancer Te-EVs.\u003c/p\u003e\u003cp\u003eAs a result of referring to RNA sequencing published by The Cancer Genome Atlas and FANTOM5, the physiological expression of TSPAN10 was localized to the retina, while almost all cases of solid tumor expressed TSPAN10 mRNA. In colorectal cancer, TSPAN10 mRNA expression was significantly associated with tumor stage and prognosis.\u003c/p\u003e\u003cp\u003eWe evaluated the diagnostic potential of TSPAN10 using serum from patients with colorectal cancer. TSPAN10 signals measured by enzyme-linked immunosorbent assay increased in a stage-dependent manner and demonstrated favorable diagnostic potential with 83% sensitivity and 84% specificity. Transmission electron microscopy showed the membrane localization of TSPAN10 on an EV derived from a patient with advanced colorectal cancer.\u003c/p\u003e\u003cp\u003eTSPAN10-expressing EVs may represent a subset of disease-associated EVs, offering diagnostic utility and advancing biological insights into EVs in cancer and other pathological conditions.\u003c/p\u003e","manuscriptTitle":"Tetraspanin-10 as a Potential Marker for Disease-Associated Extracellular Vesicles","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-09-25 23:13:59","doi":"10.21203/rs.3.rs-7690699/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":"17c01c88-c44f-4708-9250-e75336a042fe","owner":[],"postedDate":"September 25th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":55168423,"name":"Oncology"}],"tags":[],"updatedAt":"2025-09-25T23:13:59+00:00","versionOfRecord":[],"versionCreatedAt":"2025-09-25 23:13:59","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7690699","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7690699","identity":"rs-7690699","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}