Piezo1-mediated mechano-energetics regulate CAR T cell function

Piezo1-mediated mechano-energetics regulate CAR T cell function | 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 Piezo1-mediated mechano-energetics regulate CAR T cell function Weiqiang Chen, Ngoc Luu, Rui Li, Yifei Fang, Huishu Wang, Yujing Song, and 10 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7776704/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 CAR T cell cytotoxicity requires generating immense mechanical force, but the energetic costs of this process remain poorly defined. While metabolic reprogramming fuels effector function, its mechanistic connection to mechanotransduction remains unclear. By directly measuring the synaptic force and mechanical energy of single CAR T cells and linking them to their metabolic state, we proved that the mechano-energetic efficiency is a fundamental determinant of cytotoxic potency. We discovered that the mechanosensitive ion channel Piezo1 couples cytoskeletal dynamics to metabolic rewiring via Ca²⁺-Wnt-Rac1 signaling. Disrupting Piezo1 cripples glycolytic and mitochondrial ATP production, causing energetic stress and impaired cytotoxicity. Notably, Piezo1 activity follows a Goldilocks principle: intermediate level maximizes activation and cytotoxicity, whereas either hypoactive or hyperactive Piezo1 states impair mechano-metabolic fitness and drive dysfunction in patient and exhausted CAR T cells. Our work establishes mechano-metabolic coupling as a core regulator of CAR T cell fitness and pinpoints Piezo1 tuning as a new strategy to enhance cancer immunotherapy. Biological sciences/Cell biology/Cell signalling/Calcium signalling Biological sciences/Biophysics/Bioenergetics Biological sciences/Immunology/Immunotherapy Biological sciences/Biological techniques/Biophysical methods Full Text Additional Declarations Yes there is potential Competing Interest. W.C. and N.L. are inventors (New York University) on a U.S. Provisional Patent Application (63/684,051) for mechano-genetic engineering of piezo1-enhanced CAR T cells for efficient immunotherapy technology. The other authors declare no competing interests. Supplementary Files 20250929CARTSI.pdf Supplementary Information 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-7776704","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":533446617,"identity":"ce73a635-39a8-421e-9633-3bf1d265afab","order_by":0,"name":"Weiqiang 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