Opioid-expressing B Cells Silence Tumor-infiltrating Nociceptor Neurons

Opioid-expressing B Cells Silence Tumor-infiltrating Nociceptor Neurons | 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 Biological Sciences - Article Opioid-expressing B Cells Silence Tumor-infiltrating Nociceptor Neurons Sebastien Talbot, Tuany Eichwald, Maryam Ahmadi, Andre Martel Matos, and 11 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7389517/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 Nociceptor neurons, which transmit pain signals, also regulate immunity by releasing immunomodulatory neuropeptides. In head and neck squamous cell carcinoma (HNSCC) and melanoma, our research has shown that tumor-innervating nociceptors modulate anti-tumor immunity through the release of calcitonin gene-related peptide (CGRP) and its interaction with receptor activity-modifying protein 1 (RAMP1). A retrospective analysis of clinical charts from HNSCC patients revealed that higher pain levels correlated with increased opioid use, perineural invasion, and decreased B-cell infiltration—factors associated with poorer survival outcomes. In silico single-cell RNA sequencing demonstrated that opioid use in HNSCC patients downregulates nociceptin/orphanin FQ (N/OFQ), an endogenous ligand for opioid receptor-like-1 (OPRL1). We identified B cells as the primary source of N/OFQ and observed that high expression of either Pnoc or Oprl1 correlates with better survival in both melanoma and HNSCC. In a mouse model of oral squamous cell carcinoma (oSCC), we found that nociceptor neurons in tongue tumors overexpress Oprl1 and exhibit severe mechanical pain hypersensitivity. Compared to healthy tissue, oSCC tumors have dense infiltration of nociceptor fibers and N/OFQ-expressing B cells. Pharmacological blockade of Oprl1 reduced HNSCC-induced mechanical pain. In a melanoma mouse model, tumor-innervating neurons also overexpressed Oprl1, and similar overexpression was observed when DRG neurons were co-cultured with B16F10 cells. Activating OPRL1 reduced tumor size, enhanced cytotoxic T-cell infiltration, and relieved cancer-induced thermal hypersensitivity. In contrast, depleting CD19+ B cells or blocking OPRL1 led to increased tumor growth, reduced CD8+ T-cell infiltration and cytotoxic potential, exacerbated pain, and elevated CGRP levels. Moreover, we discovered that Ramp1+ B cells express Pnoc, but CGRP suppresses this expression. Blocking RAMP1 reduced tumor growth and promoted B-cell Pnoc expression. Overall, these findings suggest that targeting the N/OFQ and RAMP1 pathways could bolster anti-tumor immunity while simultaneously alleviating cancer-induced pain. Biological sciences/Cancer/Tumour immunology/Immunosurveillance Biological sciences/Neuroscience/Neuroimmunology Cancer Neuroscience Neuro-Immunology B Cells Melanoma Pain Nociceptin Head and Neck Cancer Full Text Additional Declarations There is NO Competing Interest. 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-7389517","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Biological Sciences - Article","associatedPublications":[],"authors":[{"id":507813663,"identity":"f4101408-edca-4dcd-ae8c-4f57f1a554f1","order_by":0,"name":"Sebastien Talbot","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA7UlEQVRIiWNgGAWjYFACHgjFJwEkPgAxGztxWgwY2CQYGBtngLQwk6KlGcwmpEW+vffgxx8Mf+TZpJuPP7b5tU2ej5mB8cPHHNxaDM6cS5bmYTAwbJM5ltic23fbsI2ZgVly5jY8WiRyDKSBFGObRI5hc27PbUagFjZmXjxa5Oe/Mf75g8HAHqzFsue2PUEtDDd4zCSADksEa2H4cTuRoBaDM3lp1jwGxskgv8zsbbid3MbM2IzXL/LtZw/f/FEhZ9sv3Xzgw48/t23ntzcf/PARn8MgdkFpxjYw2UBIPTL4Q4riUTAKRsEoGCkAAFxUSppSxmpnAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0001-9932-7174","institution":"Karolinska Institute","correspondingAuthor":true,"prefix":"","firstName":"Sebastien","middleName":"","lastName":"Talbot","suffix":""},{"id":507813664,"identity":"0df48a88-4145-4865-82c5-c7977b1be0aa","order_by":1,"name":"Tuany Eichwald","email":"","orcid":"","institution":"Universite de Montreal","correspondingAuthor":false,"prefix":"","firstName":"Tuany","middleName":"","lastName":"Eichwald","suffix":""},{"id":507813665,"identity":"735fc2b0-c359-45da-b8c1-c2294d481aef","order_by":2,"name":"Maryam Ahmadi","email":"","orcid":"","institution":"Department of Biomedical and Molecular Sciences, Queen’s University","correspondingAuthor":false,"prefix":"","firstName":"Maryam","middleName":"","lastName":"Ahmadi","suffix":""},{"id":507813666,"identity":"2ba81bc5-6e39-4f22-b226-99831fcd0cde","order_by":3,"name":"Andre Martel Matos","email":"","orcid":"","institution":"University of Pittsburgh","correspondingAuthor":false,"prefix":"","firstName":"Andre","middleName":"Martel","lastName":"Matos","suffix":""},{"id":507813667,"identity":"d8063b8c-039b-4884-975b-ca9309f2be89","order_by":4,"name":"Amin Reza Nikpoor","email":"","orcid":"","institution":"queen's university","correspondingAuthor":false,"prefix":"","firstName":"Amin","middleName":"Reza","lastName":"Nikpoor","suffix":""},{"id":507813668,"identity":"d0f0baaa-9611-4847-b061-52474c6c86df","order_by":5,"name":"Karine Roversi","email":"","orcid":"","institution":"Universite de Montreal","correspondingAuthor":false,"prefix":"","firstName":"Karine","middleName":"","lastName":"Roversi","suffix":""},{"id":507813669,"identity":"423becff-97b7-4e81-b6ca-6e573b8fd9b4","order_by":6,"name":"Mohammad Balood","email":"","orcid":"","institution":"Universite de Montreal","correspondingAuthor":false,"prefix":"","firstName":"Mohammad","middleName":"","lastName":"Balood","suffix":""},{"id":507813670,"identity":"00d79ed1-e874-4b6c-a32c-bf59f472a5a5","order_by":7,"name":"Fendi Obuekwe","email":"","orcid":"","institution":"University of Pittsburgh","correspondingAuthor":false,"prefix":"","firstName":"Fendi","middleName":"","lastName":"Obuekwe","suffix":""},{"id":507813671,"identity":"49788849-38a0-4bda-96f7-1b9c7ae9b1a0","order_by":8,"name":"Marci L. 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In head and neck squamous cell carcinoma (HNSCC) and melanoma, our research has shown that tumor-innervating nociceptors modulate anti-tumor immunity through the release of calcitonin gene-related peptide (CGRP) and its interaction with receptor activity-modifying protein 1 (RAMP1). A retrospective analysis of clinical charts from HNSCC patients revealed that higher pain levels correlated with increased opioid use, perineural invasion, and decreased B-cell infiltration—factors associated with poorer survival outcomes. In silico single-cell RNA sequencing demonstrated that opioid use in HNSCC patients downregulates nociceptin/orphanin FQ (N/OFQ), an endogenous ligand for opioid receptor-like-1 (OPRL1). We identified B cells as the primary source of N/OFQ and observed that high expression of either Pnoc or Oprl1 correlates with better survival in both melanoma and HNSCC. In a mouse model of oral squamous cell carcinoma (oSCC), we found that nociceptor neurons in tongue tumors overexpress Oprl1 and exhibit severe mechanical pain hypersensitivity. Compared to healthy tissue, oSCC tumors have dense infiltration of nociceptor fibers and N/OFQ-expressing B cells. Pharmacological blockade of Oprl1 reduced HNSCC-induced mechanical pain. In a melanoma mouse model, tumor-innervating neurons also overexpressed Oprl1, and similar overexpression was observed when DRG neurons were co-cultured with B16F10 cells. Activating OPRL1 reduced tumor size, enhanced cytotoxic T-cell infiltration, and relieved cancer-induced thermal hypersensitivity. In contrast, depleting CD19+ B cells or blocking OPRL1 led to increased tumor growth, reduced CD8+ T-cell infiltration and cytotoxic potential, exacerbated pain, and elevated CGRP levels. Moreover, we discovered that Ramp1+ B cells express Pnoc, but CGRP suppresses this expression. Blocking RAMP1 reduced tumor growth and promoted B-cell Pnoc expression. Overall, these findings suggest that targeting the N/OFQ and RAMP1 pathways could bolster anti-tumor immunity while simultaneously alleviating cancer-induced pain.","manuscriptTitle":"Opioid-expressing B Cells Silence Tumor-infiltrating Nociceptor Neurons","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-09-03 04:39:50","doi":"10.21203/rs.3.rs-7389517/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":"85b75d39-8f74-4f2c-a7d6-43bd1ec2cf71","owner":[],"postedDate":"September 3rd, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":54103230,"name":"Biological sciences/Cancer/Tumour immunology/Immunosurveillance"},{"id":54103231,"name":"Biological sciences/Neuroscience/Neuroimmunology"}],"tags":[],"updatedAt":"2025-10-20T14:51:15+00:00","versionOfRecord":[],"versionCreatedAt":"2025-09-03 04:39:50","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7389517","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7389517","identity":"rs-7389517","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}