Endothelial defects unveil cardiovascular phenotype in iPSC-based disease modelling across three generations of a DiGeorge syndrome family

Endothelial defects unveil cardiovascular phenotype in iPSC-based disease modelling across three generations of a DiGeorge syndrome family | 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 Endothelial defects unveil cardiovascular phenotype in iPSC-based disease modelling across three generations of a DiGeorge syndrome family Tünde Berecz, Brigitta Szabó, Ábel Fóthi, Andrea Ágnes Molnár, and 11 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7141852/v2 This work is licensed under a CC BY 4.0 License Status: Posted Version 2 posted You are reading this latest preprint version Show more versions Abstract Aims DiGeorge syndrome (DGS) due to 22q11.2 microdeletion is characterized by a high degree of phenotypic variability. This study aimed to elucidate the molecular and cellular mechanisms underlying this variability and exacerbation of cardiovascular manifestations by developing a human induced pluripotent stem cell (hiPSC)-based model using a three-generation family. Methods and results We established hiPSC lines and their cardiovascular derivatives from a family carrying the identical 22q11.2 deletion but displaying a wide spectrum of cardiovascular involvement, allowing for a direct comparison of genetics and phenotypes at the cellular level. Genetic analyses revealed no de novo mutation in the DG region genes in patients, and no correlation between the cumulative number of genetic variants and the severity of DGS. However, we observed the presence of gene variants that may modulate predisposition to the disease with possible role in cardiovascular development. Cardiomyocytes from DGS-hiPSC with prominent cardiac symptoms exhibited altered expression of connexin-43, suggesting disruptions in early cardiac morphogenesis compared to asymptomatic relatives. Endothelial cells differentiated from symptomatic DGS-hiPSC showed impaired migration and disrupted tubular morphology with dysregulation of angiogenesis and vascular integrity pathways at the transcriptomic level. To bridge in vitro and transcriptomics findings with in vivo relevance, we performed deep clinical phenotyping for each participant and applied gene-to-phenotype correlation algorithms using large-scale clinical registry data. Conclusion Our results demonstrate that hiPSC-derived cardiovascular cells from DGS patients recapitulate disease-specific morphological and functional abnormalities, enabling identification of genotype-phenotype relationships that underlie clinical heterogeneity. Translational Perspective By modelling DGS with patient-derived hiPSC from a uniquely multigenerational family, our study reveals how shared genetic microdeletions can result in divergent pathophysiological outcomes at the cellular level. This work establishes that individualised hiPSC models faithfully reflect cardiac and vascular anomalies observed in patients and delineate the cellular and transcriptional mechanisms underlying these differences. These results facilitate early identification of pathogenic mechanisms, potentially enabling more refined risk assessment and personalised monitoring strategies for DGS patients - both for cardiovascular manifestations and potentially for extra-cardiac features. Stem Cell & Developmental Cell Biology Cardiac & Cardiovascular Systems DiGeorge syndrome induced pluripotent stem cells cardiovascular disease genotype-phenotype correlation vascular biology Full Text Additional Declarations The authors declare no competing interests. Supplementary Files DiGeorgeSupplementaryFigures2025.04.01.pdf Supplementary information Berecz et al Cite Share Download PDF Status: Posted Version 2 posted You are reading this latest preprint version Show more versions 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-7141852","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":487524302,"identity":"180fc7ce-2b9b-40ac-95ee-acba94cfc657","order_by":1,"name":"Tünde Berecz","email":"","orcid":"","institution":"Institute of Molecular Life Sciences, HUN-REN RCNS, Budapest, Hungary","correspondingAuthor":false,"prefix":"","firstName":"Tünde","middleName":"","lastName":"Berecz","suffix":""},{"id":487524303,"identity":"af542a3f-3b6e-437e-87b9-13f58da4c8f6","order_by":2,"name":"Brigitta Szabó","email":"","orcid":"","institution":"Institute of Molecular Life Sciences, HUN-REN RCNS, Budapest, Hungary","correspondingAuthor":false,"prefix":"","firstName":"Brigitta","middleName":"","lastName":"Szabó","suffix":""},{"id":487524304,"identity":"0fd15b1a-c1f8-4613-913c-a1c6a563bac4","order_by":3,"name":"Ábel Fóthi","email":"","orcid":"","institution":"Institute of Molecular Life Sciences, HUN-REN RCNS, Budapest, Hungary","correspondingAuthor":false,"prefix":"","firstName":"Ábel","middleName":"","lastName":"Fóthi","suffix":""},{"id":487524305,"identity":"7af08992-a11a-49a5-83c1-d219d4ad31c4","order_by":4,"name":"Andrea Ágnes Molnár","email":"","orcid":"","institution":"Heart and Vascular Center, Semmelweis University, Budapest, Hungary","correspondingAuthor":false,"prefix":"","firstName":"Andrea","middleName":"Ágnes","lastName":"Molnár","suffix":""},{"id":487524306,"identity":"b9c47db0-a80a-4e30-ab95-f18af9caee86","order_by":5,"name":"Kristóf Árvai","email":"","orcid":"","institution":"Department of Pathology and Experimental Cancer Research, Semmelweis University, Budapest, Hungary","correspondingAuthor":false,"prefix":"","firstName":"Kristóf","middleName":"","lastName":"Árvai","suffix":""},{"id":487524307,"identity":"7e885a30-33d5-4040-bb3a-7b67e25db4bb","order_by":6,"name":"Leila Holub","email":"","orcid":"","institution":"Department of Pathology and Experimental Cancer Research, Semmelweis University, Budapest, Hungary","correspondingAuthor":false,"prefix":"","firstName":"Leila","middleName":"","lastName":"Holub","suffix":""},{"id":487524308,"identity":"b20c8c21-cf39-474a-a6d8-a8973ea893e8","order_by":7,"name":"Irén Haltrich","email":"","orcid":"","institution":"2nd Department of Pediatrics, Semmelweis University, Budapest, Hungary","correspondingAuthor":false,"prefix":"","firstName":"Irén","middleName":"","lastName":"Haltrich","suffix":""},{"id":487524309,"identity":"1eb096be-a48a-445c-aec1-c780b416e750","order_by":8,"name":"Katalin Vincze","email":"","orcid":"","institution":"Institute of Molecular Life Sciences, HUN-REN RCNS, Budapest, Hungary","correspondingAuthor":false,"prefix":"","firstName":"Katalin","middleName":"","lastName":"Vincze","suffix":""},{"id":487524310,"identity":"89116eaa-46e3-4e0d-9a2b-b0f2fc139e8d","order_by":9,"name":"György Várady","email":"","orcid":"","institution":"Institute of Molecular Life Sciences, HUN-REN RCNS, Budapest, Hungary","correspondingAuthor":false,"prefix":"","firstName":"György","middleName":"","lastName":"Várady","suffix":""},{"id":487524311,"identity":"710e79e5-8203-42a1-ae2f-10b2b5b276ee","order_by":10,"name":"Tamás I. 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This study aimed to elucidate the molecular and cellular mechanisms underlying this variability and exacerbation of cardiovascular manifestations by developing a human induced pluripotent stem cell (hiPSC)-based model using a three-generation family.\u003c/p\u003e\u003ch2\u003eMethods and results\u003c/h2\u003e\u003cp\u003eWe established hiPSC lines and their cardiovascular derivatives from a family carrying the identical 22q11.2 deletion but displaying a wide spectrum of cardiovascular involvement, allowing for a direct comparison of genetics and phenotypes at the cellular level. Genetic analyses revealed no de novo mutation in the DG region genes in patients, and no correlation between the cumulative number of genetic variants and the severity of DGS. However, we observed the presence of gene variants that may modulate predisposition to the disease with possible role in cardiovascular development. Cardiomyocytes from DGS-hiPSC with prominent cardiac symptoms exhibited altered expression of connexin-43, suggesting disruptions in early cardiac morphogenesis compared to asymptomatic relatives. Endothelial cells differentiated from symptomatic DGS-hiPSC showed impaired migration and disrupted tubular morphology with dysregulation of angiogenesis and vascular integrity pathways at the transcriptomic level. To bridge in vitro and transcriptomics findings with in vivo relevance, we performed deep clinical phenotyping for each participant and applied gene-to-phenotype correlation algorithms using large-scale clinical registry data.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e\u003cp\u003eOur results demonstrate that hiPSC-derived cardiovascular cells from DGS patients recapitulate disease-specific morphological and functional abnormalities, enabling identification of genotype-phenotype relationships that underlie clinical heterogeneity.\u003c/p\u003e\u003ch2\u003eTranslational Perspective\u003c/h2\u003e\u003cp\u003eBy modelling DGS with patient-derived hiPSC from a uniquely multigenerational family, our study reveals how shared genetic microdeletions can result in divergent pathophysiological outcomes at the cellular level. This work establishes that individualised hiPSC models faithfully reflect cardiac and vascular anomalies observed in patients and delineate the cellular and transcriptional mechanisms underlying these differences. These results facilitate early identification of pathogenic mechanisms, potentially enabling more refined risk assessment and personalised monitoring strategies for DGS patients - both for cardiovascular manifestations and potentially for extra-cardiac features.\u003c/p\u003e","manuscriptTitle":"Endothelial defects unveil cardiovascular phenotype in iPSC-based disease modelling across three generations of a DiGeorge syndrome family","msid":"","msnumber":"","nonDraftVersions":[{"code":2,"date":"2025-09-18 18:00:16","doi":"10.21203/rs.3.rs-7141852/v2","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}},{"code":1,"date":"2025-07-18 15:06:51","doi":"10.21203/rs.3.rs-7141852/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":"f602a0c7-34da-4477-a940-8c7cbc4b2212","owner":[],"postedDate":"September 18th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":51762357,"name":"Stem Cell \u0026 Developmental Cell Biology"},{"id":51762358,"name":"Cardiac \u0026 Cardiovascular Systems"}],"tags":[],"updatedAt":"2025-07-18T15:06:51+00:00","versionOfRecord":[],"versionCreatedAt":"2025-09-18 18:00:16","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v2","identity":"rs-7141852","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7141852","identity":"rs-7141852","version":["v2"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}