Targeting Artemis Sensitizes B Cells to Topoisomerase 2 Poisons by Disrupting DNA-PKcs-Dependent Repair

Targeting Artemis Sensitizes B Cells to Topoisomerase 2 Poisons by Disrupting DNA-PKcs-Dependent Repair | 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 Targeting Artemis Sensitizes B Cells to Topoisomerase 2 Poisons by Disrupting DNA-PKcs-Dependent Repair Nicholas Pannunzio, Melissa Folkerts, Cameron Hom, Angie Nguyen, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6317145/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 Topoisomerase 2 (Top2) poisons are widely used in cancer therapy but are associated with toxicity and secondary malignancies. Removing Top2 adducts requires endonuclease activity and repair by non-homologous end joining (NHEJ). We show that the NHEJ enzyme Artemis is a promising target for co-treatment with Top2 poisons. Inhibition of the Artemis activator, DNA-PKcs, with peposertib (M3814) sensitizes B cells to Top2 poisons while ATM or ATR inhibition does not. Interestingly, while M3814 treatment blocks Artemis endonuclease activity, Artemis phosphorylation is still detectible and is only affected upon inhibiting ATM, suggestive of an additional role for Artemis in DNA damage response signaling. Additionally, Artemis loss results in a significant accumulation of Top2 DNA adducts following treatment, indicating Artemis may act outside its canonical role in NHEJ to reduce adduct burden. Clinical data demonstrates that high Artemis expression correlates with poor survival in several cancers, and we demonstrate that Artemis function is critical for survival following combination drug treatment. These insights can be leveraged to unlock new avenues for the treatment of aggressive cancers by enhancing the cytotoxicity of agents through blockade of DNA break repair. Biological sciences/Cancer/Cancer genetics Biological sciences/Molecular biology/DNA damage and repair/Non-homologous-end joining NHEJ DNA repair Artemis topoisomerase poison DNA-PKcs ATM B cell peposertib (M3814) Full Text Additional Declarations There is NO Competing Interest. Supplementary Files FolkertsetalSupplementaryInformation.pdf Supplementary Data 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-6317145","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":440026689,"identity":"8e3ca31b-afd9-4642-8e78-3e00978ba23b","order_by":0,"name":"Nicholas Pannunzio","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAu0lEQVRIiWNgGAWjYPACmwQwBSQZG4jUkka6lsMJMBZhLebSZw9+LtxxPk++gcfwwwMGG9kNBwhosezLS5aeeeZ2scEBHmOJBIY0Y4JaDM7wGEjztt1O3MDAYwb0y+FEYrQY/+ZtO5c4vwGs5T9RWsyAthxIbDgA1nKAGC18ada8bcmJGw6zFUskGCQbzySshffwbd42u8T57c0bP/6osJPtI6SFgYEHSjODTSCoHFnLKBgFo2AUjAJcAACYLD+r5wkpDQAAAABJRU5ErkJggg==","orcid":"https://orcid.org/0000-0002-8290-8236","institution":"University of Califormia Irvine","correspondingAuthor":true,"prefix":"","firstName":"Nicholas","middleName":"","lastName":"Pannunzio","suffix":""},{"id":440026690,"identity":"295d86ca-a191-48c2-8bd4-60543330d0ca","order_by":1,"name":"Melissa Folkerts","email":"","orcid":"https://orcid.org/0009-0009-9657-6146","institution":"University of Califormia Irvine","correspondingAuthor":false,"prefix":"","firstName":"Melissa","middleName":"","lastName":"Folkerts","suffix":""},{"id":440026691,"identity":"db0b3ccb-d4fc-4868-9907-7cf8f8614038","order_by":2,"name":"Cameron Hom","email":"","orcid":"","institution":"University of Califormia Irvine","correspondingAuthor":false,"prefix":"","firstName":"Cameron","middleName":"","lastName":"Hom","suffix":""},{"id":440026692,"identity":"3b49839c-9b69-4913-b421-1e70fa324c94","order_by":3,"name":"Angie Nguyen","email":"","orcid":"https://orcid.org/0000-0002-9200-3510","institution":"University of Califormia Irvine","correspondingAuthor":false,"prefix":"","firstName":"Angie","middleName":"","lastName":"Nguyen","suffix":""},{"id":440026693,"identity":"b424dd0d-f799-4b15-9818-784acfe56418","order_by":4,"name":"Kaiyuan Shen","email":"","orcid":"https://orcid.org/0009-0007-1224-8811","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Kaiyuan","middleName":"","lastName":"Shen","suffix":""},{"id":440026694,"identity":"9d3d0fa3-67b1-4cd0-b608-d7bab5030a16","order_by":5,"name":"Valeria Rangel","email":"","orcid":"","institution":"University of Califormia Irvine","correspondingAuthor":false,"prefix":"","firstName":"Valeria","middleName":"","lastName":"Rangel","suffix":""},{"id":440026695,"identity":"a7bbee85-8c62-4199-a72b-a55bd7204726","order_by":6,"name":"Pedro Ortega","email":"","orcid":"https://orcid.org/0000-0003-4216-3695","institution":"University of California, Irvine","correspondingAuthor":false,"prefix":"","firstName":"Pedro","middleName":"","lastName":"Ortega","suffix":""},{"id":440026696,"identity":"fdf153e8-cf15-452e-a008-46d93092af91","order_by":7,"name":"Rémi Buisson","email":"","orcid":"https://orcid.org/0000-0002-7196-8209","institution":"University of California, Irvine","correspondingAuthor":false,"prefix":"","firstName":"Rémi","middleName":"","lastName":"Buisson","suffix":""},{"id":440026697,"identity":"3624ff14-4ace-4631-80bf-b22baabe5cf3","order_by":8,"name":"Selma Masri","email":"","orcid":"https://orcid.org/0000-0002-8619-8331","institution":"University of Califormia Irvine","correspondingAuthor":false,"prefix":"","firstName":"Selma","middleName":"","lastName":"Masri","suffix":""},{"id":440026698,"identity":"23cb35fd-bb88-4217-887d-231042a3dd3b","order_by":9,"name":"Noritaka Adachi","email":"","orcid":"https://orcid.org/0000-0002-1895-0747","institution":"Yokohama City Univ, Japan","correspondingAuthor":false,"prefix":"","firstName":"Noritaka","middleName":"","lastName":"Adachi","suffix":""},{"id":440026699,"identity":"c3cbd177-9f12-48ca-baaa-dcc868f47db6","order_by":10,"name":"Katheryn Meek","email":"","orcid":"https://orcid.org/0000-0002-0454-4414","institution":"Michigan State University","correspondingAuthor":false,"prefix":"","firstName":"Katheryn","middleName":"","lastName":"Meek","suffix":""}],"badges":[],"createdAt":"2025-03-27 05:40:13","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6317145/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6317145/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":84960736,"identity":"3de0aa11-b49f-4541-b0e0-2bec0366d722","added_by":"auto","created_at":"2025-06-19 09:03:43","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1078906,"visible":true,"origin":"","legend":"Article File","description":"","filename":"FolkerselalArtemis.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6317145/v1_covered_f44add42-1a39-420a-921c-bf570466172c.pdf"},{"id":84960090,"identity":"c451f4bf-3346-4a09-aabc-d63d92ddcdc2","added_by":"auto","created_at":"2025-06-19 08:55:40","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":1348298,"visible":true,"origin":"","legend":"Supplementary Data","description":"","filename":"FolkertsetalSupplementaryInformation.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6317145/v1/4eadba1b746f9853fdf6542d.pdf"}],"financialInterests":"There is \u003cb\u003eNO\u003c/b\u003e Competing Interest.","formattedTitle":"Targeting Artemis Sensitizes B Cells to Topoisomerase 2 Poisons by Disrupting DNA-PKcs-Dependent Repair","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"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":"nature-portfolio","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"","title":"Nature Portfolio","twitterHandle":"","acdcEnabled":false,"dfaEnabled":false,"editorialSystem":"ejp","reportingPortfolio":"","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"NHEJ, DNA repair, Artemis, topoisomerase poison, DNA-PKcs, ATM, B cell, peposertib (M3814)","lastPublishedDoi":"10.21203/rs.3.rs-6317145/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6317145/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Topoisomerase 2 (Top2) poisons are widely used in cancer therapy but are associated with toxicity and secondary malignancies. Removing Top2 adducts requires endonuclease activity and repair by non-homologous end joining (NHEJ). We show that the NHEJ enzyme Artemis is a promising target for co-treatment with Top2 poisons. Inhibition of the Artemis activator, DNA-PKcs, with peposertib (M3814) sensitizes B cells to Top2 poisons while ATM or ATR inhibition does not. Interestingly, while M3814 treatment blocks Artemis endonuclease activity, Artemis phosphorylation is still detectible and is only affected upon inhibiting ATM, suggestive of an additional role for Artemis in DNA damage response signaling. Additionally, Artemis loss results in a significant accumulation of Top2 DNA adducts following treatment, indicating Artemis may act outside its canonical role in NHEJ to reduce adduct burden. Clinical data demonstrates that high Artemis expression correlates with poor survival in several cancers, and we demonstrate that Artemis function is critical for survival following combination drug treatment. These insights can be leveraged to unlock new avenues for the treatment of aggressive cancers by enhancing the cytotoxicity of agents through blockade of DNA break repair.","manuscriptTitle":"Targeting Artemis Sensitizes B Cells to Topoisomerase 2 Poisons by Disrupting DNA-PKcs-Dependent Repair","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-06-19 08:55:35","doi":"10.21203/rs.3.rs-6317145/v1","editorialEvents":[],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"communications-biology","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"commsbio","sideBox":"Learn more about [Communications Biology](http://www.nature.com/commsbio/)","snPcode":"","submissionUrl":"","title":"Communications Biology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Communications Series","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"31cd2689-83a3-4a3b-8528-d202f2ff0228","owner":[],"postedDate":"June 19th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":46844075,"name":"Biological sciences/Cancer/Cancer genetics"},{"id":46844076,"name":"Biological sciences/Molecular biology/DNA damage and repair/Non-homologous-end joining"}],"tags":[],"updatedAt":"2026-04-14T10:56:53+00:00","versionOfRecord":[],"versionCreatedAt":"2025-06-19 08:55:35","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6317145","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6317145","identity":"rs-6317145","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}