Oxidative damage reprograms the Hippo-WNT network via X-linked Kdm6a to activate blastocyst dormancy and prevent offspring tumorigenesis.

Oxidative damage reprograms the Hippo-WNT network via X-linked Kdm6a to activate blastocyst dormancy and prevent offspring tumorigenesis. | 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 Oxidative damage reprograms the Hippo-WNT network via X-linked Kdm6a to activate blastocyst dormancy and prevent offspring tumorigenesis. zhiling Li, Yue Huang, Siyao Ha, Xueru Wang, Youyin Pang, Jiliang Huang, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6242185/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 vitro fertilization (IVF) has been associated with an increased risk of tumorigenesis in offspring. Our previous research indicated that oxidative damage-induced X-chromosome aneuploidy (XCA) in IVF mouse embryos may contribute to tumorigenesis in offspring. However, the tumorigenic mechanisms underlying this phenomenon remain unclear. The present study elucidates that an elevated number of X chromosomes leads to excessive transcription of Xist , resulting in aberrant X-chromosome inactivation (XCI). This abnormal XCI subsequently inhibits the expression of the X-linked lysine demethylase 6A (Kdm6a), which is followed by an increase in repressive marker H3K27me3 and a decrease in active markers H3K27ac/H3K4me3. To investigate the epigenetic mechanisms involved in offspring tumorigenesis, we employed CUT&Tag technology to map genome-wide profiles of H3K27ac/H3K4me3/H3K27me3 in IVF blastocysts. We found that Kdm6a-dependent histone modifications exhibited a close relationship with leukemia by regulating cancer pathways, particularly Hippo/Yap1 and Wnt (Wnt/β-catenin and Wnt/RhoA) signalings in oxidatively damaged IVF embryos. Kdm6a plasmid and antioxidant EGCG were found to maintain epigenetic stability and antagonize the effects of ROS on the Hippo and Wnt pathways. We concluded that oxidative damage-induced loss of Kdm6a participated in offspring tumorigenesis via oncogenic RhoA/β-catenin activation and tumor-suppressive Hippo inactivation during IVF. However, leukemia or hepatic tumors was not increased in the offspring derived from oxidatively damaged IVF blastocysts. Our further analysis revealed that Kdm6a-dependent histone modifications play a crucial role in regulating pluripotency of embryonic stem cells. Kdm6a plasmid antagonized the effects of ROS on the formation of rosette-like structures and the expression of naive gene Oct4 as well as primed gene Otx2 in IVF blastocysts. This implies that Kdm6a is essentail for the naive-to-primed transition and activation of blastocysts during implantation. We hypothesize that loss of Kdm6a adversely affects blastocyst implantation, thereby to prevent the birth of offspring with an increased risk of tumorigenesis. Biological sciences/Developmental biology/Embryogenesis/Cell lineage Biological sciences/Molecular biology/Epigenetics/DNA methylation Biological sciences/Molecular biology/DNA damage and repair/DNA damage response Biological sciences/Cancer/Cancer prevention In vitro fertilization oxidative damage X chromosome inactivation X-linked Kdm6a Hippo/Wnt pathways offspring tumorigenesis blastocyst activation Full Text Additional Declarations There is NO Competing Interest. Supplemental tables 1 and 2 are not available with this version. 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-6242185","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":431009585,"identity":"f805aa86-796a-4cf1-abce-8b5ad5c50455","order_by":0,"name":"zhiling Li","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA20lEQVRIiWNgGAWjYFAC5oYDjA0SMmwMDIwPEipsiNDBxgjWwgPUwmzw4EwacVoYgIgHxJR82HaIsA75+Y2NB3/usODhk26/VpHAdoCBv707Aa8Wg2NAh0meATpM5kzZjQSeOwwSZ85uwK8F5BfDNqAWiZy0GwkSzxgMJHLxa5FvA2pJhGopSDA4TFgLA8hhB8Fa0o8xJCQQocXgWGLDwUaILcwSCQfSeAj6Rb758OGPP9vq5ORnpD/8+POfjRx/ey8BhyEAjwGYJFY5CLA/IEX1KBgFo2AUjCAAAPKzSSRw44BrAAAAAElFTkSuQmCC","orcid":"","institution":"The First Affiliated Hospital of Shantou University Medical College","correspondingAuthor":true,"prefix":"","firstName":"zhiling","middleName":"","lastName":"Li","suffix":""},{"id":431009586,"identity":"9fbd0325-7057-486e-9ec4-55be34fc6e7f","order_by":1,"name":"Yue Huang","email":"","orcid":"https://orcid.org/0000-0002-1120-0580","institution":"The First Affiliated Hospital of Shantou University Medical College","correspondingAuthor":false,"prefix":"","firstName":"Yue","middleName":"","lastName":"Huang","suffix":""},{"id":431009587,"identity":"79c2c3c2-98c0-41e5-9490-cb92215f900c","order_by":2,"name":"Siyao Ha","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Siyao","middleName":"","lastName":"Ha","suffix":""},{"id":431009588,"identity":"48f93b0d-6db4-4369-9ebf-ebc357b76855","order_by":3,"name":"Xueru Wang","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Xueru","middleName":"","lastName":"Wang","suffix":""},{"id":431009589,"identity":"80c95d6c-7a20-4be5-a39b-61aba3d550f0","order_by":4,"name":"Youyin Pang","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Youyin","middleName":"","lastName":"Pang","suffix":""},{"id":431009590,"identity":"816dd449-43ca-4ed6-b1e3-80c385d15a7a","order_by":5,"name":"Jiliang Huang","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Jiliang","middleName":"","lastName":"Huang","suffix":""},{"id":431009591,"identity":"4d61fce7-0bae-493b-ae26-b6ed7f39fcc2","order_by":6,"name":"Lihong Liu","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Lihong","middleName":"","lastName":"Liu","suffix":""},{"id":431009592,"identity":"a6fa0f31-b8c4-4749-bfa8-16338c096a28","order_by":7,"name":"Gaizhen Ru","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Gaizhen","middleName":"","lastName":"Ru","suffix":""},{"id":431009593,"identity":"2c306093-0d78-4380-b3d9-1e821c878bc3","order_by":8,"name":"Siyi Peng","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Siyi","middleName":"","lastName":"Peng","suffix":""}],"badges":[],"createdAt":"2025-03-17 07:51:05","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6242185/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6242185/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":79078150,"identity":"d03c8927-ce9a-4b3e-a838-501c3125d3a5","added_by":"auto","created_at":"2025-03-24 07:47:34","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3728577,"visible":true,"origin":"","legend":"Article File","description":"","filename":"344090artfile885661st85hj.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6242185/v1_covered_a833baa3-6834-4e0c-be15-cdadc45546db.pdf"}],"financialInterests":"\u003cp\u003eThere is \u003cstrong\u003eNO\u003c/strong\u003e Competing Interest.\u003c/p\u003e\n\u003cp\u003eSupplemental tables 1 and 2 are not available with this version.\u003c/p\u003e","formattedTitle":"Oxidative damage reprograms the Hippo-WNT network via X-linked Kdm6a to activate blastocyst dormancy and prevent offspring tumorigenesis.","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"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":"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},"keywords":"In vitro fertilization, oxidative damage, X chromosome inactivation, X-linked Kdm6a, Hippo/Wnt pathways, offspring tumorigenesis, blastocyst activation","lastPublishedDoi":"10.21203/rs.3.rs-6242185/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6242185/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cem\u003eIn vitro \u003c/em\u003efertilization (IVF) has been associated with an increased risk of tumorigenesis in offspring. Our previous research indicated that oxidative damage-induced X-chromosome aneuploidy (XCA) in IVF mouse embryos may contribute to tumorigenesis in offspring. However, the tumorigenic mechanisms underlying this phenomenon remain unclear. The present study elucidates that an elevated number of X chromosomes leads to excessive transcription of \u003cem\u003eXist\u003c/em\u003e, resulting in aberrant X-chromosome inactivation (XCI). This abnormal XCI subsequently inhibits the expression of the X-linked lysine demethylase 6A (Kdm6a), which is followed by an increase in repressive marker H3K27me3 and a decrease in active markers H3K27ac/H3K4me3. To investigate the epigenetic mechanisms involved in offspring tumorigenesis, we employed CUT\u0026amp;Tag technology to map genome-wide profiles of H3K27ac/H3K4me3/H3K27me3 in IVF blastocysts. We found that Kdm6a-dependent histone modifications exhibited a close relationship with leukemia by regulating cancer pathways, particularly Hippo/Yap1 and Wnt (Wnt/β-catenin and Wnt/RhoA) signalings in oxidatively damaged IVF embryos. Kdm6a plasmid and antioxidant EGCG were found to maintain epigenetic stability and antagonize the effects of ROS on the Hippo and Wnt pathways. We concluded that oxidative damage-induced loss of Kdm6a participated in offspring tumorigenesis via oncogenic RhoA/β-catenin activation and tumor-suppressive Hippo inactivation during IVF. However, leukemia or hepatic tumors was not increased in the offspring derived from oxidatively damaged IVF blastocysts. Our further analysis revealed that Kdm6a-dependent histone modifications play a crucial role in regulating pluripotency of embryonic stem cells. Kdm6a plasmid antagonized the effects of ROS on the formation of rosette-like structures and the expression of naive gene \u003cem\u003eOct4\u003c/em\u003e as well as primed gene \u003cem\u003eOtx2\u003c/em\u003e in IVF blastocysts. This implies that Kdm6a is essentail for the naive-to-primed transition and activation of blastocysts during implantation. We hypothesize that loss of Kdm6a adversely affects blastocyst implantation, thereby to prevent the birth of offspring with an increased risk of tumorigenesis.\u003c/p\u003e","manuscriptTitle":"Oxidative damage reprograms the Hippo-WNT network via X-linked Kdm6a to activate blastocyst dormancy and prevent offspring tumorigenesis.","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-03-24 07:15:26","doi":"10.21203/rs.3.rs-6242185/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":"9de1acff-1f35-41d5-a52e-304c87fef069","owner":[],"postedDate":"March 24th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":45906191,"name":"Biological sciences/Developmental biology/Embryogenesis/Cell lineage"},{"id":45906192,"name":"Biological sciences/Molecular biology/Epigenetics/DNA methylation"},{"id":45906193,"name":"Biological sciences/Molecular biology/DNA damage and repair/DNA damage response"},{"id":45906194,"name":"Biological sciences/Cancer/Cancer prevention"}],"tags":[],"updatedAt":"2025-03-24T07:15:28+00:00","versionOfRecord":[],"versionCreatedAt":"2025-03-24 07:15:26","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6242185","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6242185","identity":"rs-6242185","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}