NuMA1 controls myonuclear motility in striated skeletal muscle through AMPK activity and is impaired in Duchenne Muscular Dystrophy

NuMA1 controls myonuclear motility in striated skeletal muscle through AMPK activity and is impaired in Duchenne Muscular Dystrophy | 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 NuMA1 controls myonuclear motility in striated skeletal muscle through AMPK activity and is impaired in Duchenne Muscular Dystrophy Vincent Gache, Nathalie Couturier, Léa Castellano, Alireza Ghasemizadeh, and 12 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7713210/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 4 You are reading this latest preprint version Abstract Skeletal muscle consists in a bundle of thousands post-mitotic multinucleated cells, called myofibers, in which myonuclei are evenly spaced and positioned at the periphery. This myonuclear positioning i) shapes myonuclear domain (MND) in myofibers, essential for the transcriptional integrity of myofibers, ii) is driven by cytoskeleton and associated proteins and iii) is required for proper myofiber functions. In numerous of muscle diseases (i.e. myopathies), alteration of myonuclei localization (internalized and/or mispositioned) contributes to myofiber misfunctioning, supporting the need to better understand the fundamental mechanisms that regulates myonuclei dynamics and to restore the establishment of myonuclear domains in these diseases. In this study, we show that in Duchenne muscular dystrophy (DMD) myofibers, myonuclei are more dynamic and contribute to the failure in MNDs settings, suggesting that in pathological conditions enhanced myonuclear motility impact myonuclear distribution. To identify new actors regulating this MND settings, we performed a mass spectrometry (MS)-based proteomic analysis to identify microtubule associated proteins (MAPs) in myotubes/myofibers and performed a siRNA screening on selected candidates. This approach allowed to highlight NuMA1 as a new factor controlling myoblast fusion and myonuclear positioning through the control of nuclear-microtubule-organizing-center (n-MTOC) integrity and microtubule network orientation. Strikingly, while NuMA1 is restrained to myonuclei in mononucleated myoblasts, it progressively accumulates in the cytoplasm during muscle cell differentiation, preferentially with microtubule (MT) nucleation spots at the vicinity of the nuclear membrane. We identified that AMP Kinase activity has an essential role in NuMA1 nuclear accumulation through the specific phosphorylation on serine-1853 and the ability of myonuclei to accumulate NuMA1 is correlated to their motility in myofibers. Finally, we show that nuclear NuMA1 content is increased in DMD patients and mdx mouse model, contributing to more dynamic myonuclei that can manipulated pharmacologically through the control of AMPK activity. Altogether, our data identify a novel mechanism by which nuclear sequestration of a MAP allows to couple nuclear positioning and motion to MT organization. Biological sciences/Cell biology/Cytoskeleton/Microtubules Biological sciences/Developmental biology/Differentiation Biological sciences/Cell biology/Nuclear organization/Nucleoskeleton Biological sciences/Developmental biology/Disease model Biological sciences/Cell biology/Mechanisms of disease Full Text Additional Declarations There is NO Competing Interest. Cite Share Download PDF Status: Under Review Version 1 posted Unknown event 16 Oct, 2025 Editorial decision: Reject before peer review 14 Oct, 2025 Editor assigned by journal 25 Sep, 2025 First submitted to journal 25 Sep, 2025 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-7713210","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":520614926,"identity":"5118d1f8-6e14-4917-9690-4a9754cc8f0e","order_by":0,"name":"Vincent 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This myonuclear positioning i) shapes myonuclear domain (MND) in myofibers, essential for the transcriptional integrity of myofibers, ii) is driven by cytoskeleton and associated proteins and iii) is required for proper myofiber functions. In numerous of muscle diseases (i.e. myopathies), alteration of myonuclei localization (internalized and/or mispositioned) contributes to myofiber misfunctioning, supporting the need to better understand the fundamental mechanisms that regulates myonuclei dynamics and to restore the establishment of myonuclear domains in these diseases. In this study, we show that in Duchenne muscular dystrophy (DMD) myofibers, myonuclei are more dynamic and contribute to the failure in MNDs settings, suggesting that in pathological conditions enhanced myonuclear motility impact myonuclear distribution. To identify new actors regulating this MND settings, we performed a mass spectrometry (MS)-based proteomic analysis to identify microtubule associated proteins (MAPs) in myotubes/myofibers and performed a siRNA screening on selected candidates. This approach allowed to highlight NuMA1 as a new factor controlling myoblast fusion and myonuclear positioning through the control of nuclear-microtubule-organizing-center (n-MTOC) integrity and microtubule network orientation. Strikingly, while NuMA1 is restrained to myonuclei in mononucleated myoblasts, it progressively accumulates in the cytoplasm during muscle cell differentiation, preferentially with microtubule (MT) nucleation spots at the vicinity of the nuclear membrane. We identified that AMP Kinase activity has an essential role in NuMA1 nuclear accumulation through the specific phosphorylation on serine-1853 and the ability of myonuclei to accumulate NuMA1 is correlated to their motility in myofibers. Finally, we show that nuclear NuMA1 content is increased in DMD patients and mdx mouse model, contributing to more dynamic myonuclei that can manipulated pharmacologically through the control of AMPK activity. Altogether, our data identify a novel mechanism by which nuclear sequestration of a MAP allows to couple nuclear positioning and motion to MT organization.","manuscriptTitle":"NuMA1 controls myonuclear motility in striated skeletal muscle through AMPK activity and is impaired in Duchenne Muscular Dystrophy","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-09-29 02:46:38","doi":"10.21203/rs.3.rs-7713210/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"transferred","content":"Cell Death \u0026 Differentiation","date":"2025-10-16T15:04:20+00:00","index":"","fulltext":""},{"type":"decision","content":"Reject before peer review","date":"2025-10-14T15:45:14+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-09-25T13:38:14+00:00","index":"","fulltext":""},{"type":"submitted","content":"Nature Cell Biology","date":"2025-09-25T12:53:29+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"cell-death-and-disease","isNatureJournal":false,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"cddis","sideBox":"Learn more about [Cell Death \u0026 Disease](http://www.nature.com/cddis/)","snPcode":"41419","submissionUrl":"https://mts-cddis.nature.com/cgi-bin/main.plex","title":"Cell Death \u0026 Disease","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Nature AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"f51f91a8-d2af-4a86-be71-bf7bdf3fb024","owner":[],"postedDate":"September 29th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":55340351,"name":"Biological sciences/Cell biology/Cytoskeleton/Microtubules"},{"id":55340352,"name":"Biological sciences/Developmental biology/Differentiation"},{"id":55340353,"name":"Biological sciences/Cell biology/Nuclear organization/Nucleoskeleton"},{"id":55340354,"name":"Biological sciences/Developmental biology/Disease model"},{"id":55340355,"name":"Biological sciences/Cell biology/Mechanisms of disease"}],"tags":[],"updatedAt":"2025-10-28T17:05:00+00:00","versionOfRecord":[],"versionCreatedAt":"2025-09-29 02:46:38","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7713210","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7713210","identity":"rs-7713210","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}