Single-cell transcriptomics reveal the crosstalk between platelet-associated TRAF4high endothelial cell subpopulation and tumor microenvironment in colorectal cancer

Single-cell transcriptomics reveal the crosstalk between platelet-associated TRAF4high endothelial cell subpopulation and tumor microenvironment in colorectal cancer | 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 Single-cell transcriptomics reveal the crosstalk between platelet-associated TRAF4high endothelial cell subpopulation and tumor microenvironment in colorectal cancer Guoqing Su, Nannan Wang, Yuepeng Cao, Jinghang Qian, Yi Wang, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4506500/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 Background Platelet-tumor endothelial cell interactions are pivotal in the colorectal cancer microenvironment, driving tumor invasion, metastasis and shaping the immune landscape. Investigating these interactions sheds light on the underlying mechanisms of colorectal cancer progression and opens avenues for novel therapeutic interventions. Methods By interrogating 10x Genomics single-cell transcriptomic data from 23 publicly available samples, we constructed a paired cancerous and adjacent non-cancerous single-cell atlas for colorectal cancer (CRC). Utilizing a comprehensive suite of analyses, including area under the curve (AUC), copy number variation, pseudotime trajectory, enrichment, and intercellular communication assessments, we delineated the heterogeneity within CRC endothelial cells. Furthermore, our SCENIC transcription factor analysis pinpointed the most active transcription factors within the platelet-associated endothelial cell subpopulation. The functional relevance of TRAF4 in endothelial cell was subsequently confirmed by in vitro validation and large cohort clinical samples. Results We delineated a distinct subpopulation of endothelial cells, designated as C2, that is profoundly influenced by platelet education and is intricately linked to tumor metastasis, adverse prognosis, and immune cell infiltration. This subpopulation exhibits a strong association with the immune cell infiltration characteristic of the tumor microenvironment. Notably, TRAF4 emerged as a pivotal platelet-related transcription factors within this subgroup, with its downregulation leading to diminished migration, angiogenic potential, and chemotactic responsiveness of endothelial cells to CD8 + T cells. Additionally, the expression of TRAF4 and its correlation with CD8 + T cell infiltration were corroborated in clinical specimens. Conclusions Our investigation has identified a platelet-associated subpopulation of tumor endothelial cells that contributes to the suppression of CD8 + T cell infiltration within the tumor microenvironment and promotes tumor angiogenesis, thereby facilitating malignant tumor progression. Furthermore, we have pinpointed TRAF4 as a pivotal transcription factor within this subpopulation, underscoring its significant promise as an innovative therapeutic target for CRC. colorectal cancer platelet endothelial cell TRAF4 CD8 + T cell Single-cell transcriptomics metastasis Full Text Supplementary Files SupplementaryFile1.csv SupplementaryFile2.csv SupplementaryFile3.csv SupplementaryFile4.csv SupplementaryFile5.csv SupplementaryFile6.csv SupplementaryTable1.pdf Supplementaryfigures.docx 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. 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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-4506500","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":321029990,"identity":"920b5242-2764-46c0-822c-4c8872c3da94","order_by":0,"name":"Guoqing Su","email":"","orcid":"","institution":"Jiangsu Cancer Institute and Hospital: Jiangsu Cancer Hospital","correspondingAuthor":false,"prefix":"","firstName":"Guoqing","middleName":"","lastName":"Su","suffix":""},{"id":321029991,"identity":"e1703712-82fd-47e6-b1f5-1678612fc841","order_by":1,"name":"Nannan Wang","email":"","orcid":"","institution":"Jiangsu Cancer Institute and 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cancer","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":"colorectal cancer, platelet, endothelial cell, TRAF4, CD8 + T cell, Single-cell transcriptomics, metastasis","lastPublishedDoi":"10.21203/rs.3.rs-4506500/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4506500/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e \u003cp\u003ePlatelet-tumor endothelial cell interactions are pivotal in the colorectal cancer microenvironment, driving tumor invasion, metastasis and shaping the immune landscape. Investigating these interactions sheds light on the underlying mechanisms of colorectal cancer progression and opens avenues for novel therapeutic interventions.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e \u003cp\u003eBy interrogating 10x Genomics single-cell transcriptomic data from 23 publicly available samples, we constructed a paired cancerous and adjacent non-cancerous single-cell atlas for colorectal cancer (CRC). Utilizing a comprehensive suite of analyses, including area under the curve (AUC), copy number variation, pseudotime trajectory, enrichment, and intercellular communication assessments, we delineated the heterogeneity within CRC endothelial cells. Furthermore, our SCENIC transcription factor analysis pinpointed the most active transcription factors within the platelet-associated endothelial cell subpopulation. The functional relevance of TRAF4 in endothelial cell was subsequently confirmed by in vitro validation and large cohort clinical samples.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eWe delineated a distinct subpopulation of endothelial cells, designated as C2, that is profoundly influenced by platelet education and is intricately linked to tumor metastasis, adverse prognosis, and immune cell infiltration. This subpopulation exhibits a strong association with the immune cell infiltration characteristic of the tumor microenvironment. Notably, TRAF4 emerged as a pivotal platelet-related transcription factors within this subgroup, with its downregulation leading to diminished migration, angiogenic potential, and chemotactic responsiveness of endothelial cells to CD8\u0026thinsp;+\u0026thinsp;T cells. Additionally, the expression of TRAF4 and its correlation with CD8\u0026thinsp;+\u0026thinsp;T cell infiltration were corroborated in clinical specimens.\u003c/p\u003e\u003ch2\u003eConclusions\u003c/h2\u003e \u003cp\u003eOur investigation has identified a platelet-associated subpopulation of tumor endothelial cells that contributes to the suppression of CD8\u0026thinsp;+\u0026thinsp;T cell infiltration within the tumor microenvironment and promotes tumor angiogenesis, thereby facilitating malignant tumor progression. Furthermore, we have pinpointed TRAF4 as a pivotal transcription factor within this subpopulation, underscoring its significant promise as an innovative therapeutic target for CRC.\u003c/p\u003e","manuscriptTitle":"Single-cell transcriptomics reveal the crosstalk between platelet-associated TRAF4high endothelial cell subpopulation and tumor microenvironment in colorectal cancer","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-07-23 13:45:12","doi":"10.21203/rs.3.rs-4506500/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":"443e1ab9-9f94-4fc9-9587-71f970ce7e32","owner":[],"postedDate":"July 23rd, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-07-23T15:10:06+00:00","versionOfRecord":[],"versionCreatedAt":"2024-07-23 13:45:12","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4506500","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4506500","identity":"rs-4506500","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}