A Combined Bioinformatics and Clinical Validation Approach Identifies CEACAM1 and RRAS2 as Key SASP-Related Biomarkers in Neuropathic Pain

A Combined Bioinformatics and Clinical Validation Approach Identifies CEACAM1 and RRAS2 as Key SASP-Related Biomarkers in Neuropathic Pain | 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 A Combined Bioinformatics and Clinical Validation Approach Identifies CEACAM1 and RRAS2 as Key SASP-Related Biomarkers in Neuropathic Pain Lulu Yan, Junlong Yang, Xiaoping Shen, Hongguang Lu This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8471120/v1 This work is licensed under a CC BY 4.0 License Status: Under Revision Version 1 posted 13 You are reading this latest preprint version Abstract Background: Neuropathic pain (NP) represents a persistent pain syndrome that develops following injury to or pathological alterations in the somatosensory pathways. Inflammatory signaling associated with cellular senescence has recently been proposed as an important factor in the development of NP. However, the precise molecular underpinnings of this relationship remain poorly defined. Methods: Differential gene expression analysis of the public dataset GSE150408 (dorsal root ganglia from a rodent neuropathic pain model) identified differentially expressed genes (DEGs). Candidate genes were identified through cross-referencing with the SASP gene collection documented in the SenMayo repository. Two computational approaches were employed for feature selection: the Least Absolute Shrinkage and Selection Operator (LASSO) regression model alongside Support Vector Machine-Recursive Feature Elimination (SVM-RFE) methodology, which collectively narrowed down the gene candidates. Subsequently, a comprehensive analytical framework was applied, which included constructing a diagnostic nomogram, performing gene set enrichment analysis (GSEA), profiling immune cell infiltration (via CIBERSORT), and conducting in silico drug prediction. Experimental verification of biomarker expression levels was conducted via quantitative reverse transcription polymerase chain reaction (RT-qPCR) analysis. The validation utilized peripheral blood mononuclear cells (PBMCs) obtained from an independent sample cohort consisting of five NP patients and five age-matched healthy individuals serving as controls. Between-group comparisons were performed through unpaired two-tailed Student's t-test to determine statistical significance. Results: Carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) and Ras-related protein R-Ras2 (RRAS2) were identified as core SASP-related biomarkers for NP. A diagnostic nomogram incorporating these two genes exhibited strong discriminatory power, achieving an area under the curve (AUC) of 0.92. Subsequent experimental validation in human blood samples confirmed a significant upregulation of CEACAM1 and concomitant downregulation of RRAS2 in NP patients compared to healthy controls ( p <0.05), thereby corroborating our initial computational predictions. Conclusion: Combining computational biology with clinical validation, this study definitively identifies CEACAM1 and RRAS2 as robust SASP-associated biomarkers for NP. Our findings not only shed light on the intricate interplay between cellular senescence and neuroinflammation in NP pathogenesis but also provide a foundational framework for developing novel diagnostic strategies and targeted therapies. The identified biomarkers demonstrate considerable potential for enhancing diagnostic and therapeutic strategies in managing this clinically challenging condition. Neuropathic pain (NP) Senescence-associated secretory phenotype (SASP) Biomarker Neuroinflammation CEACAM1 RRAS2 Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Revision Version 1 posted Editorial decision: Revision requested 26 Jan, 2026 Reviews received at journal 25 Jan, 2026 Reviews received at journal 21 Jan, 2026 Reviewers agreed at journal 19 Jan, 2026 Reviewers agreed at journal 17 Jan, 2026 Reviews received at journal 16 Jan, 2026 Reviewers agreed at journal 16 Jan, 2026 Reviewers agreed at journal 15 Jan, 2026 Reviewers agreed at journal 14 Jan, 2026 Reviewers invited by journal 14 Jan, 2026 Editor assigned by journal 13 Jan, 2026 Submission checks completed at journal 13 Jan, 2026 First submitted to journal 29 Dec, 2025 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-8471120","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":576165315,"identity":"effabc48-f579-478b-b4b4-81d3fd642754","order_by":0,"name":"Lulu Yan","email":"","orcid":"","institution":"Affiliated Hospital of Guizhou Medical University","correspondingAuthor":false,"prefix":"","firstName":"Lulu","middleName":"","lastName":"Yan","suffix":""},{"id":576165316,"identity":"5755365d-9084-4107-8b81-3655a2681558","order_by":1,"name":"Junlong Yang","email":"","orcid":"","institution":"Affiliated Hospital of Guizhou Medical 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Pain","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"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":"molecular-neurobiology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"moln","sideBox":"Learn more about [Molecular Neurobiology](https://www.springer.com/journal/12035)","snPcode":"12035","submissionUrl":"https://submission.nature.com/new-submission/12035/3","title":"Molecular Neurobiology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer 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Inflammatory signaling associated with cellular senescence has recently been proposed as an important factor in the development of NP. However, the precise molecular underpinnings of this relationship remain poorly defined.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods:\u003c/strong\u003e Differential gene expression analysis of the public dataset GSE150408 (dorsal root ganglia from a rodent neuropathic pain model) identified differentially expressed genes (DEGs). Candidate genes were identified through cross-referencing with the SASP gene collection documented in the SenMayo repository. Two computational approaches were employed for feature selection: the Least Absolute Shrinkage and Selection Operator (LASSO) regression model alongside Support Vector Machine-Recursive Feature Elimination (SVM-RFE) methodology, which collectively narrowed down the gene candidates. Subsequently, a comprehensive analytical framework was applied, which included constructing a diagnostic nomogram, performing gene set enrichment analysis (GSEA), profiling immune cell infiltration (via CIBERSORT), and conducting \u003cem\u003ein silico\u003c/em\u003e drug prediction. Experimental verification of biomarker expression levels was conducted via quantitative reverse transcription polymerase chain reaction (RT-qPCR) analysis. The validation utilized peripheral blood mononuclear cells (PBMCs) obtained from an independent sample cohort consisting of five NP patients and five age-matched healthy individuals serving as controls. Between-group comparisons were performed through unpaired two-tailed Student's t-test to determine statistical significance.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults:\u003c/strong\u003e Carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) and Ras-related protein R-Ras2 (RRAS2) were identified as core SASP-related biomarkers for NP. A diagnostic nomogram incorporating these two genes exhibited strong discriminatory power, achieving an area under the curve (AUC) of 0.92. Subsequent experimental validation in human blood samples confirmed a significant upregulation of CEACAM1 and concomitant downregulation of RRAS2 in NP patients compared to healthy controls (\u003cem\u003ep\u003c/em\u003e\u0026lt;0.05), thereby corroborating our initial computational predictions.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion:\u003c/strong\u003e Combining computational biology with clinical validation, this study definitively identifies CEACAM1 and RRAS2 as robust SASP-associated biomarkers for NP. Our findings not only shed light on the intricate interplay between cellular senescence and neuroinflammation in NP pathogenesis but also provide a foundational framework for developing novel diagnostic strategies and targeted therapies. The identified biomarkers demonstrate considerable potential for enhancing diagnostic and therapeutic strategies in managing this clinically challenging condition.\u003c/p\u003e","manuscriptTitle":"A Combined Bioinformatics and Clinical Validation Approach Identifies CEACAM1 and RRAS2 as Key SASP-Related Biomarkers in Neuropathic Pain","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-01-19 14:23:35","doi":"10.21203/rs.3.rs-8471120/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-01-26T19:04:16+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-01-25T21:50:57+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-01-21T13:41:36+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"148036339315071461974450165857999329081","date":"2026-01-20T03:46:18+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"273816014901798573263518648961906175795","date":"2026-01-17T16:54:41+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-01-17T03:52:39+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"305024401673228969550995473978267281772","date":"2026-01-17T03:25:31+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"137506844277200537481366680821948877163","date":"2026-01-15T09:21:10+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"138482873814707935527991227048022965245","date":"2026-01-15T04:08:31+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-01-15T03:20:51+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-01-13T09:03:53+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-01-13T08:56:54+00:00","index":"","fulltext":""},{"type":"submitted","content":"Molecular Neurobiology","date":"2025-12-29T08:46:31+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"molecular-neurobiology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"moln","sideBox":"Learn more about [Molecular Neurobiology](https://www.springer.com/journal/12035)","snPcode":"12035","submissionUrl":"https://submission.nature.com/new-submission/12035/3","title":"Molecular Neurobiology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"555f6b16-b3ac-45ce-a89d-06e188d4c1f9","owner":[],"postedDate":"January 19th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"in-revision","subjectAreas":[],"tags":[],"updatedAt":"2026-04-21T15:38:14+00:00","versionOfRecord":[],"versionCreatedAt":"2026-01-19 14:23:35","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8471120","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8471120","identity":"rs-8471120","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}