Inhibiting sirtuin-dependent DNA repair and oxidative stress responses impairs DIPG cell survival

Inhibiting sirtuin-dependent DNA repair and oxidative stress responses impairs DIPG cell survival | 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 Research Article Inhibiting sirtuin-dependent DNA repair and oxidative stress responses impairs DIPG cell survival Sarah A. King, Rana Rheem, Kathryn M. Spitler, Brianne R. O’Leary, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7723497/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 8 You are reading this latest preprint version Abstract Background : Diffuse intrinsic pontine gliomas (DIPG) are universally fatal pediatric brain tumors often characterized by a H3 histone mutation (H3K27M) that leads to epigenetic alterations and gene dysregulation. Histone deacetylases (HDAC), key players in modulating epigenetic pathways in cancer, have emerged as potential therapeutic targets for DIPG. This study evaluates the class III HDAC inhibitor, sirtinol, as a potential therapeutic for DIPG. Methods : Sirtinol’s efficacy and cellular mechanism were investigated using two patient-derived DIPG cell lines and one immortalized normal human astrocyte (NHAi) cell line as a comparator. Clonogenic survival was assessed under different experimental conditions, and changes in protein expression were measured via western blot and enzymatic assays. Liquid chromatography tandem mass spectrometry (LC-MS-MS) was used to measure brain and blood penetrance of sirtinol in mice. Results : We found that sirtinol dose-dependently decreased clonogenic cell survival in DIPG cells while having negligible effects on NHAi cells. Combined treatment of sirtinol and radiation resulted in additive toxicity in DIPG, but not NHAi cells. Sirtinol was less toxic in DIPG cells overexpressing mitochondrial-targeted catalase, suggesting peroxide generation as one mode of cell killing. Moreover, sirtinol was found to increase DNA double strand breaks in DIPG cells and decrease phosphorylation of SIRT2-mediated DNA repair pathway kinase ataxia-telangiectasia and Rad3-related protein (ATR). Sirtinol was detected in the brains of sirtinol-treated mice, suggesting blood brain barrier penetrance. Conclusions : Based on our results, sirtinol shows promise as a selective, redox-modulating therapeutic for DIPG that enhances oxidative stress and interferes with DNA repair. Pediatric Brain Cancer Oxidative Stress DNA Damage DIPG H3K27M Histone Deacetylase Inhibitor Sirtinol Glioma Full Text Additional Declarations No competing interests reported. Supplementary Files SupplementalFig.1.tif SupplementalFigure2UncroppedWesternsFinal.pdf Cite Share Download PDF Status: Under Review Version 1 posted Reviews received at journal 14 Nov, 2025 Reviewers agreed at journal 12 Nov, 2025 Reviewers agreed at journal 12 Nov, 2025 Reviewers invited by journal 11 Nov, 2025 Editor invited by journal 11 Nov, 2025 Editor assigned by journal 08 Oct, 2025 Submission checks completed at journal 08 Oct, 2025 First submitted to journal 08 Oct, 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-7723497","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":547177988,"identity":"50682d2e-e15c-4367-a94d-c3abf44addc7","order_by":0,"name":"Sarah A. 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survival\u003c/p\u003e","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"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":"bmc-cancer","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"bcan","sideBox":"Learn more about [BMC Cancer](http://bmccancer.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/bcan/default.aspx","title":"BMC Cancer","twitterHandle":"BMC_series","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Pediatric Brain Cancer, Oxidative Stress, DNA Damage, DIPG, H3K27M, Histone Deacetylase Inhibitor, Sirtinol, Glioma","lastPublishedDoi":"10.21203/rs.3.rs-7723497/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7723497/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground\u003c/strong\u003e: Diffuse intrinsic pontine gliomas (DIPG) are universally fatal pediatric brain tumors often characterized by a H3 histone mutation (H3K27M) that leads to epigenetic alterations and gene dysregulation. Histone deacetylases (HDAC), key players in modulating epigenetic pathways in cancer, have emerged as potential therapeutic targets for DIPG. \u0026nbsp;This study evaluates the class III HDAC inhibitor, sirtinol, as a potential therapeutic for DIPG.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods\u003c/strong\u003e: Sirtinol’s efficacy and cellular mechanism were investigated using two patient-derived DIPG cell lines and one immortalized normal human astrocyte (NHAi) cell line as a comparator. Clonogenic survival was assessed under different experimental conditions, and changes in protein expression were measured via western blot and enzymatic assays. Liquid chromatography tandem mass spectrometry (LC-MS-MS) was used to measure brain and blood penetrance of sirtinol in mice.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults\u003c/strong\u003e: We found that sirtinol dose-dependently decreased clonogenic cell survival in DIPG cells while having negligible effects on NHAi cells. Combined treatment of sirtinol and radiation resulted in additive toxicity in DIPG, but not NHAi cells. Sirtinol was less toxic in DIPG cells overexpressing mitochondrial-targeted catalase, suggesting peroxide generation as one mode of cell killing. Moreover, sirtinol was found to increase DNA double strand breaks in DIPG cells and decrease phosphorylation of SIRT2-mediated DNA repair pathway kinase ataxia-telangiectasia and Rad3-related protein (ATR). Sirtinol was detected in the brains of sirtinol-treated mice, suggesting blood brain barrier penetrance.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusions\u003c/strong\u003e: Based on our results, sirtinol shows promise as a selective, redox-modulating therapeutic for DIPG that enhances oxidative stress and interferes with DNA repair.\u003c/p\u003e","manuscriptTitle":"Inhibiting sirtuin-dependent DNA repair and oxidative stress responses impairs DIPG cell survival","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-11-21 06:22:22","doi":"10.21203/rs.3.rs-7723497/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"editorInvitedReview","content":"","date":"2025-11-14T22:23:39+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"62608772480560787556066758021669575470","date":"2025-11-12T18:02:52+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"285160750363453160736643688493232685300","date":"2025-11-12T16:55:42+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-11-11T16:33:07+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2025-11-11T15:48:14+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-10-08T18:06:46+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-10-08T13:48:11+00:00","index":"","fulltext":""},{"type":"submitted","content":"BMC Cancer","date":"2025-10-08T13:46:31+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"bmc-cancer","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"bcan","sideBox":"Learn more about [BMC Cancer](http://bmccancer.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/bcan/default.aspx","title":"BMC Cancer","twitterHandle":"BMC_series","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"6396c6c9-bf39-4c8e-b006-c54414c737c7","owner":[],"postedDate":"November 21st, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2025-11-21T06:22:22+00:00","versionOfRecord":[],"versionCreatedAt":"2025-11-21 06:22:22","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7723497","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7723497","identity":"rs-7723497","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}