Oxygen Delivery Reprograms Hypoxic Tumor Niche to Rejuvenate CAR-NKT Cells for Improved Cancer Immunotherapy

Oxygen Delivery Reprograms Hypoxic Tumor Niche to Rejuvenate CAR-NKT Cells for Improved Cancer Immunotherapy | 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 Oxygen Delivery Reprograms Hypoxic Tumor Niche to Rejuvenate CAR-NKT Cells for Improved Cancer Immunotherapy Song Li, Parvaiz A. Shiekh, Yan-Ruide Li, Haochen Nan, Lili Yang, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8296891/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted You are reading this latest preprint version Abstract Adoptive cell therapies have revolutionized the treatment of hematologic malignancies, but their success in solid tumors remains limited. Chimeric antigen receptor (CAR)-engineered invariant natural killer T (CAR-NKT) cells represent a promising frontier in immunotherapy, combining innate immune properties with engineered specificity against cancer. Despite their potential, the therapeutic efficacy of CAR lymphocytes is often compromised by tumor hypoxia. A hallmark of solid tumors, hypoxia reprograms immune cell metabolism, promotes immunosuppression, drives exhaustion and impairs cytotoxicity. To address this, we introduce Hypoxia-attenuating Oxygenating Platform for Enhanced-Immunotherapy (HOPE), an oxygen delivering modular platform that alleviates hypoxic tumor microenvironment and enhances CAR-NKT cell functionality in vivo . Using in vitro hypoxia model, we demonstrate how HOPE reverses hypoxia-induced suppression, restores mitochondrial membrane potential, synergizes CAR-NKT cell functionality, and reduces immune exhaustion. Furthermore, in vivo xenograft solid tumor model shows that HOPE increases intratumoral CAR-NKT persistence and significantly reduces tumor burden. Together, these results establish HOPE as a translatable immunoengineering strategy that rejuvenates CAR-NKT activation, metabolism and checkpoint phenotype and improves antitumor efficacy in hypoxic solid tumors, which has broad potential as an enabling platform for next-generation adoptive cell therapies. Biological sciences/Cancer/Cancer therapy/Cancer immunotherapy Physical sciences/Engineering/Biomedical engineering Invariant natural Killer T (NKT) cells hypoxia induced exhaustion chimeric antigen receptor (CAR) tumor microenvironment reprogramming cancer immunotherapy Full Text Additional Declarations Yes there is potential Competing Interest. P.A.S., Y.-R.L., L.Y. and S.L. are filing a patent application as inventors. L.Y. is a scientific advisor to AlzChem and Amberstone Biosciences, and a co-founder, stockholder, and advisory board member of Appia Bio. None of the declared companies contributed to or directed any of the research reported in this article. The other authors declare no competing interests. Supplementary Files Supplementaryinformation.docx Supplementart Information Cite Share Download PDF Status: Under Review 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-8296891","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":607499386,"identity":"75d25689-6437-46e1-8b61-944d3df0f2dc","order_by":0,"name":"Song Li","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAu0lEQVRIiWNgGAWjYLCCBwY2DAzsIBYbsVoSDNIYGJhJ08JwmAQt8u29h18kFJxP7G9mfsDwoewwYS0GZ86lWSQY3E6ccZjNgHHGOWK0SOSYGYC0bGBmMGDmbSNCi/wMsJZzQC3sH5j/EqOF4UaO8YMEgwNALTwGzIzEaDE4c8YMGMjJxjMO8xQc7DmXToTD2nuMP3z4Yyfb396+8cGPMmsiHAaMCwkY6wBR6oGA+QOxKkfBKBgFo2CEAgBzSTnSi4KsOAAAAABJRU5ErkJggg==","orcid":"https://orcid.org/0000-0002-4760-8828","institution":"University of California, Los Angeles","correspondingAuthor":true,"prefix":"","firstName":"Song","middleName":"","lastName":"Li","suffix":""},{"id":607499387,"identity":"3d183f18-53dd-4907-b324-eb500707a943","order_by":1,"name":"Parvaiz A. 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Chimeric antigen receptor (CAR)-engineered invariant natural killer T (CAR-NKT) cells represent a promising frontier in immunotherapy, combining innate immune properties with engineered specificity against cancer. Despite their potential, the therapeutic efficacy of CAR lymphocytes is often compromised by tumor hypoxia. A hallmark of solid tumors, hypoxia reprograms immune cell metabolism, promotes immunosuppression, drives exhaustion and impairs cytotoxicity. To address this, we introduce Hypoxia-attenuating Oxygenating Platform for Enhanced-Immunotherapy (HOPE), an oxygen delivering modular platform that alleviates hypoxic tumor microenvironment and enhances CAR-NKT cell functionality \u003cem\u003ein vivo\u003c/em\u003e. Using in vitro hypoxia model, we demonstrate how HOPE reverses hypoxia-induced suppression, restores mitochondrial membrane potential, synergizes CAR-NKT cell functionality, and reduces immune exhaustion. Furthermore, \u003cem\u003ein vivo\u003c/em\u003e xenograft solid tumor model shows that HOPE increases intratumoral CAR-NKT persistence and significantly reduces tumor burden. 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