Intermittent fasting reverses T cell lymphopenia via inosine to mitigate brain tumors

Intermittent fasting reverses T cell lymphopenia via inosine to mitigate brain tumors | 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 Article Intermittent fasting reverses T cell lymphopenia via inosine to mitigate brain tumors Yucai Wang, Chengkun Ye, Yang Liu, Qiuhong Zhu, Fangzhen Li, Qin Wang, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6080923/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 Brain tumors are characterized by a paucity of tumor-infiltrating T cells and systemic T cell lymphopenia, both of which contribute to immunotherapy resistance. While dietary restriction modulates immunity, how intermittent fasting (IF) reshapes T cell dynamics in brain tumors remains unknown. Here, we demonstrate that IF reverses T cell lymphopenia by liberating T cells sequestered in the bone marrow, enabling their infiltration into brain tumors and synergizing with anti-PD-1 therapy. Strikingly, this redistribution occurs specifically during the refeeding phase, which is marked by purine metabolic reprogramming and an elevation in circulating inosine levels. Mechanistically, inosine activates adenosine receptor (AR)-PKA signaling in T cells, driving phosphorylation and subsequent lysosomal degradation of CXCR4, the chemokine receptor responsible for bone marrow sequestration. Administering inosine alone recapitulates IF’s effects, restoring peripheral T cell levels and enhancing intratumoral T cell infiltration. Combined with anti-PD-1, inosine achieves robust tumor suppression in glioma and brain metastasis models (breast and lung cancer). Our work uncovers a transformative link between dietary cycles and systemic immunity: fasting-refeeding metabolic fluctuations reprogram T cell trafficking via a purine metabolite, overcoming local and systemic immunosuppression. These insights position inosine as a readily translatable therapeutic strategy to augment immunotherapy efficacy in brain tumors and beyond. Health sciences/Oncology/Cancer/CNS cancer Biological sciences/Cancer/CNS cancer Full Text Additional Declarations There is NO Competing Interest. Supplementary Files ExtendedData.pdf 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-6080923","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":422427862,"identity":"7ce1b2eb-2475-4a27-9c52-3271ac00ed38","order_by":0,"name":"Yucai Wang","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA5UlEQVRIiWNgGAWjYLCCBBDB3gDlHSBaC88BqGqitICBRAKRWgxuJD/88LDNLk8+8o3x5w81DHJ8NxIYPxfg1ZJmLJHYllxseDvHTOLAMQZjyRsJzNIz8GrJYZBI3MacuHF2jhnDwQaGxA03EtiYefBrYf6RuK0+cePMM8YfgFrqidHCBrTlcOJ8CR4DCaCWBANCWiTPPDOzSPx3PHEDT1qZxJljEoYzzzxslsanhe948uObP85UJ85vP7z5Q0WNjTxQ5OBnfFoUDsBcCGFIADFjAx4NDAzyDeiMUTAKRsEoGAXoAAAyXFOEI7Bo3AAAAABJRU5ErkJggg==","orcid":"","institution":"University of Science and Technology of China","correspondingAuthor":true,"prefix":"","firstName":"Yucai","middleName":"","lastName":"Wang","suffix":""},{"id":422427863,"identity":"fc8897ad-3137-43d6-ba72-f20b07dccfb3","order_by":1,"name":"Chengkun Ye","email":"","orcid":"","institution":"The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, 230001, China","correspondingAuthor":false,"prefix":"","firstName":"Chengkun","middleName":"","lastName":"Ye","suffix":""},{"id":422427864,"identity":"a536afcd-59d4-4391-8349-e4c7fcb020d3","order_by":2,"name":"Yang Liu","email":"","orcid":"","institution":"The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, 230001, China","correspondingAuthor":false,"prefix":"","firstName":"Yang","middleName":"","lastName":"Liu","suffix":""},{"id":422427865,"identity":"f00fcfe0-5137-46d6-9a1e-5aaa0fee807b","order_by":3,"name":"Qiuhong Zhu","email":"","orcid":"","institution":"University of Science and Technology of China","correspondingAuthor":false,"prefix":"","firstName":"Qiuhong","middleName":"","lastName":"Zhu","suffix":""},{"id":422427866,"identity":"f7a02a15-7d78-4754-b549-f90a4a2d26b8","order_by":4,"name":"Fangzhen Li","email":"","orcid":"","institution":"University of Science and Technology of China","correspondingAuthor":false,"prefix":"","firstName":"Fangzhen","middleName":"","lastName":"Li","suffix":""},{"id":422427867,"identity":"c73befe9-8718-4e41-a1b3-766508d55c01","order_by":5,"name":"Qin Wang","email":"","orcid":"","institution":"The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, 230001, China","correspondingAuthor":false,"prefix":"","firstName":"Qin","middleName":"","lastName":"Wang","suffix":""},{"id":422427868,"identity":"ab444324-9a68-4c73-bfe7-21048cd0a62b","order_by":6,"name":"Niu Chaoshi","email":"","orcid":"","institution":"The First Affiliated Hospital of USTC","correspondingAuthor":false,"prefix":"","firstName":"Niu","middleName":"","lastName":"Chaoshi","suffix":""},{"id":422427869,"identity":"10b35774-fd8a-4ded-b977-a759d99b1b61","order_by":7,"name":"Wenchao Zhou","email":"","orcid":"https://orcid.org/0000-0002-6650-1831","institution":"University of Science and Technology of China","correspondingAuthor":false,"prefix":"","firstName":"Wenchao","middleName":"","lastName":"Zhou","suffix":""}],"badges":[],"createdAt":"2025-02-21 15:59:12","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6080923/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6080923/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":78753267,"identity":"294cb294-ebcc-4bff-b744-9b1249cb07a1","added_by":"auto","created_at":"2025-03-18 12:15:16","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1367839,"visible":true,"origin":"","legend":"Article File","description":"","filename":"Manuscript20250221.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6080923/v1_covered_d47bce7b-c71c-41dc-b046-8ffff5f3bc83.pdf"},{"id":78752365,"identity":"1c94a37b-37e1-491a-a51f-56cb0360f1a3","added_by":"auto","created_at":"2025-03-18 12:07:11","extension":"pdf","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":1072820,"visible":true,"origin":"","legend":"","description":"","filename":"ExtendedData.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6080923/v1/a368481aa89ece9c51dd2937.pdf"}],"financialInterests":"There is \u003cb\u003eNO\u003c/b\u003e Competing Interest.","formattedTitle":"Intermittent fasting reverses T cell lymphopenia via inosine to mitigate brain tumors","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"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":"","lastPublishedDoi":"10.21203/rs.3.rs-6080923/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6080923/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Brain tumors are characterized by a paucity of tumor-infiltrating T cells and systemic T cell lymphopenia, both of which contribute to immunotherapy resistance. While dietary restriction modulates immunity, how intermittent fasting (IF) reshapes T cell dynamics in brain tumors remains unknown. Here, we demonstrate that IF reverses T cell lymphopenia by liberating T cells sequestered in the bone marrow, enabling their infiltration into brain tumors and synergizing with anti-PD-1 therapy. Strikingly, this redistribution occurs specifically during the refeeding phase, which is marked by purine metabolic reprogramming and an elevation in circulating inosine levels. Mechanistically, inosine activates adenosine receptor (AR)-PKA signaling in T cells, driving phosphorylation and subsequent lysosomal degradation of CXCR4, the chemokine receptor responsible for bone marrow sequestration. Administering inosine alone recapitulates IF’s effects, restoring peripheral T cell levels and enhancing intratumoral T cell infiltration. Combined with anti-PD-1, inosine achieves robust tumor suppression in glioma and brain metastasis models (breast and lung cancer). Our work uncovers a transformative link between dietary cycles and systemic immunity: fasting-refeeding metabolic fluctuations reprogram T cell trafficking via a purine metabolite, overcoming local and systemic immunosuppression. These insights position inosine as a readily translatable therapeutic strategy to augment immunotherapy efficacy in brain tumors and beyond.","manuscriptTitle":"Intermittent fasting reverses T cell lymphopenia via inosine to mitigate brain tumors","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-03-18 12:07:06","doi":"10.21203/rs.3.rs-6080923/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":"fd94e84f-5fa6-4beb-bcbb-55be05ba3c6e","owner":[],"postedDate":"March 18th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":45030956,"name":"Health sciences/Oncology/Cancer/CNS cancer"},{"id":45030957,"name":"Biological sciences/Cancer/CNS cancer"}],"tags":[],"updatedAt":"2025-03-18T12:07:06+00:00","versionOfRecord":[],"versionCreatedAt":"2025-03-18 12:07:06","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6080923","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6080923","identity":"rs-6080923","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}