Trained to repair: repeated muscle injury imprints a ‘healthy regenerative memory’

Trained to repair: repeated muscle injury imprints a ‘healthy regenerative memory’ | 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 Trained to repair: repeated muscle injury imprints a ‘healthy regenerative memory’ Leonit Kiriaev, Chrystal Tiong, Hannah Huckstep, Kathrin Mattes, and 8 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9585345/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 In healthy skeletal muscle, eccentric exercise confers protection against damage during subsequent bouts; the repeated bout effect. Repeated cycles of degeneration and regeneration drive pathological remodelling during ageing and in neuromuscular diseases such as Duchenne muscular dystrophy. Whether repeated injury in healthy muscle drives cumulative degradation or encodes a beneficial state remains unresolved. We tested whether repeated muscle injury and regeneration produce a mechanically resilient rather than progressively compromised phenotype. Adult male C57BL/10 mice underwent one (1X) or three (3X) rounds of myotoxic injury alongside undamaged controls and dystrophic mice, assessed using ex vivo contractile physiology, histology, RNA sequencing, and proteomics. Repeated injury produced persistent structural remodelling including reduced fibre cross-sectional area, increased mass, branching and fibre-type redistribution. Despite this, muscles exhibited preserved isometric force, resistance to eccentric damage, and superior recovery exceeding singly injured and mdx muscles . Multi omics profiling revealed extensive remodelling, characterised by ECM expansion, reduced oxidative metabolism, and widespread post transcriptional regulation in immune and calcium handling pathways. These findings define a healthy regenerative memory: a stably remodelled, mechanically resilient state induced by repeated regeneration, offering a molecular basis for distinguishing adaptive from pathological remodelling with relevance to the repeat bout effect in exercise physiology and neuromuscular disease. Health sciences/Neurology/Neurological disorders/Neuromuscular disease Health sciences/Anatomy/Musculoskeletal system/Muscle/Skeletal muscle Biological sciences/Physiology Biological sciences/Biotechnology/Regenerative medicine Biological sciences/Developmental biology/Self-renewal Full Text Additional Declarations There is NO Competing Interest. Supplementary Files Supplementary2.jpg Supplementary Figure S2 Supplementary1.jpg Supplementary Figure S1 Supplementary4.jpg Supplementary Figure S4 Supplementary3.jpg Supplementary Figure S3 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-9585345","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":632943494,"identity":"cf1a7d77-d041-4cf2-bf3c-b34ad3426eb9","order_by":0,"name":"Leonit 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Repeated cycles of degeneration and regeneration drive pathological remodelling during ageing and in neuromuscular diseases such as Duchenne muscular dystrophy. Whether repeated injury in healthy muscle drives cumulative degradation or encodes a beneficial state remains unresolved.\r\nWe tested whether repeated muscle injury and regeneration produce a mechanically resilient rather than progressively compromised phenotype. Adult male C57BL/10 mice underwent one (1X) or three (3X) rounds of myotoxic injury alongside undamaged controls and dystrophic mice, assessed using \u003ci\u003eex vivo\u003c/i\u003e contractile physiology, histology, RNA sequencing, and proteomics.\r\nRepeated injury produced persistent structural remodelling including reduced fibre cross-sectional area, increased mass, branching and fibre-type redistribution. Despite this, muscles exhibited preserved isometric force, resistance to eccentric damage, and superior recovery exceeding singly injured and \u003ci\u003emdx muscles\u003c/i\u003e. Multi omics profiling revealed extensive remodelling, characterised by ECM expansion, reduced oxidative metabolism, and widespread post transcriptional regulation in immune and calcium handling pathways.\r\nThese findings define a healthy regenerative memory: a stably remodelled, mechanically resilient state induced by repeated regeneration, offering a molecular basis for distinguishing adaptive from pathological remodelling with relevance to the repeat bout effect in exercise physiology and neuromuscular disease.","manuscriptTitle":"Trained to repair: repeated muscle injury imprints a ‘healthy regenerative memory’","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-05-04 18:24:12","doi":"10.21203/rs.3.rs-9585345/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":"6adda657-6610-4f5f-be49-d63e7a1592ab","owner":[],"postedDate":"May 4th, 2026","published":true,"recentEditorialEvents":[{"type":"decision","content":"Reject before peer review","date":"2026-05-07T12:34:42+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-05-05T18:07:04+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-05-01T11:54:17+00:00","index":"","fulltext":""},{"type":"submitted","content":"Nature Communications","date":"2026-05-01T10:54:38+00:00","index":"","fulltext":""}],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":67371175,"name":"Health sciences/Neurology/Neurological disorders/Neuromuscular disease"},{"id":67371176,"name":"Health sciences/Anatomy/Musculoskeletal system/Muscle/Skeletal muscle"},{"id":67371177,"name":"Biological sciences/Physiology"},{"id":67371178,"name":"Biological sciences/Biotechnology/Regenerative medicine"},{"id":67371179,"name":"Biological sciences/Developmental biology/Self-renewal"}],"tags":[],"updatedAt":"2026-05-07T12:36:23+00:00","versionOfRecord":[],"versionCreatedAt":"2026-05-04 18:24:12","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-9585345","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-9585345","identity":"rs-9585345","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}