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Gene-Edited Human-Induced Pluripotent Stem Cell Lines to Elucidate DAND5 Function throughout Cardiac Differentiation

2023-02-05Artigo científico Acesso aberto
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dc.contributor.authorInácio, José M
dc.contributor.authorNunes, Mafalda M
dc.contributor.authorAlmeida, Micael
dc.contributor.authorCristo, Fernando
dc.contributor.authorAnjos, Rui
dc.contributor.authorBelo, José A
dc.contributor.authorA. Belo, José
dc.contributor.institutionNOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM)
dc.contributor.institutioniNOVA4Health - pólo NMS
dc.contributor.pblMDPI - Multidisciplinary Digital Publishing Institute
dc.date.accessioned2023-03-15T22:29:31Z
dc.date.available2023-03-15T22:29:31Z
dc.date.issued2023-02-05
dc.descriptionFunding: This work was supported by Fundação para a Ciência e a Tecnologia (PTDC/ BIMMED/3363/2014) and Scientific Employment Stimulus to J.M.I. (Norma Transitória 8189/2018), predoctoral fellowship to M.A. (FCT; 06890/2021/BD), post-doctoral fellowship to F.C. (DAI 2019/08/ SAICTPAC/0047/2015) and iNOVA4Health – UIDB/04462/2020 and UIDP/04462/2020, two programs financially supported by Fundação para a Ciência e Tecnologia/ Ministério da Ciência, Tecnologia e Ensino Superior.
dc.description.abstract(1) Background: The contribution of gene-specific variants for congenital heart disease, one of the most common congenital disabilities, is still far from our complete understanding. Here, we applied a disease model using human-induced pluripotent stem cells (hiPSCs) to evaluate the function of DAND5 on human cardiomyocyte (CM) differentiation and proliferation. (2) Methods: Taking advantage of our DAND5 patient-derived iPSC line, we used CRISPR-Cas9 gene-editing to generate a set of isogenic hiPSCs (DAND5-corrected and DAND5 full-mutant). The hiPSCs were differentiated into CMs, and RT-qPCR and immunofluorescence profiled the expression of cardiac markers. Cardiomyocyte proliferation was analysed by flow cytometry. Furthermore, we used a multi-electrode array (MEA) to study the functional electrophysiology of DAND5 hiPSC-CMs. (3) Results: The results indicated that hiPSC-CM proliferation is affected by DAND5 levels. Cardiomyocytes derived from a DAND5 full-mutant hiPSC line are more proliferative when compared with gene-corrected hiPSC-CMs. Moreover, parallel cardiac differentiations showed a differential cardiac gene expression profile, with upregulated cardiac progenitor markers in DAND5-KO hiPSC-CMs. Microelectrode array (MEA) measurements demonstrated that DAND5-KO hiPSC-CMs showed prolonged field potential duration and increased spontaneous beating rates. In addition, conduction velocity is reduced in the monolayers of hiPSC-CMs with full-mutant genotype. (4) Conclusions: The absence of DAND5 sustains the proliferation of hiPSC-CMs, which alters their electrophysiological maturation properties. These results using DAND5 hiPSC-CMs consolidate the findings of the in vitro and in vivo mouse models, now in a translational perspective. Altogether, the data will help elucidate the molecular mechanism underlying this human heart disease and potentiates new therapies for treating adult CHD.en
dc.description.versionpublishersversion
dc.description.versionpublished
dc.format.extent1354876
dc.identifier.doi10.3390/cells12040520
dc.identifier.issn2073-4409
dc.identifier.otherPURE: 54519507
dc.identifier.otherPURE UUID: d30f4a0d-53c0-4bba-9231-a2b9234a2ce6
dc.identifier.otherPubMed: 36831187
dc.identifier.otherPubMedCentral: PMC9954670
dc.identifier.otherScopus: 85148850894
dc.identifier.urihttp://hdl.handle.net/10362/150629
dc.language.isoeng
dc.peerreviewedyes
dc.subjectDAND5
dc.subjectcardiomyocyte proliferation
dc.subjectcongenital heart disease
dc.subjectdisease modelling
dc.subjectSDG 3 - Good Health and Well-being
dc.titleGene-Edited Human-Induced Pluripotent Stem Cell Lines to Elucidate DAND5 Function throughout Cardiac Differentiationen
dc.typejournal article
degois.publication.issue4
degois.publication.titleCells
degois.publication.volume12
dspace.entity.typePublication
person.familyNameBelo
person.givenNameJosé A.
person.identifier.ciencia-idBF13-08E9-25E6
person.identifier.orcid0000-0001-7384-0949
person.identifier.ridF-4444-2012
person.identifier.scopus-author-id6602141392
rcaap.rightsopenAccess
relation.isAuthorOfPublication9f75224c-9cf2-478f-9c7c-d22527950642
relation.isAuthorOfPublication.latestForDiscovery9f75224c-9cf2-478f-9c7c-d22527950642

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