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Generation of Cellular Models for Fabry Disease: Unlocking the Potential of iPSCs and Gene Editing

2025-11-20Documento de conferência Acesso aberto
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datacite.subject.fosCiências Médicas::Outras Ciências Médicas
datacite.subject.sdg03:Saúde de Qualidade
dc.contributor.authorDuarte, Ana Joana
dc.contributor.authorMoreira, Luciana
dc.contributor.authorRibeiro, Diogo
dc.contributor.authorAlves, Sandra
dc.contributor.authorGaspar, Paulo
dc.contributor.authorBragança, José
dc.contributor.authorAmaral, Olga
dc.date.accessioned2026-03-04T11:42:07Z
dc.date.available2026-03-04T11:42:07Z
dc.date.issued2025-11-20
dc.descriptionAbstract available
dc.description.abstractIntroduction: Fabry Disease (FD) is a lysosomal storage disorder caused by mutations in the GLA gene, resulting in a defective α-GAL A enzyme. This deficiency leads to the accumulation of Gb3 and lyso-Gb3 within lysosomes, resulting in a multisystem disease. Through reprogramming, we obtained induced pluripotent stem cells (iPSCs) derived from fibroblasts of a patient with FD2 and from a wild-type (WT) control. We used CRISPR/Cas9 to correct the c.860G>A mutation present in the patient’s cells, as well as to generate a WT GLA knockout (KO). The resulting cells were then differentiated into cardiomyocytes, a cell type affected by this disease. Methods: We reprogrammed the fibroblasts into iPSCs using episomal vectors or Sendai virus. For gene editing, single-guide RNAs (sgRNAs) and Cas9 were nucleofected, and the editing was confirmed by Sanger sequencing. Following colony selection, isogenic cell lines were established. The FD iPSCs, the corrected FD iPSCs, and the WT iPSCs were then differentiated into iPSC-derived cardiomyocytes (iPSC-CMs). Results: Seven new cell models were generated. Functional studies of the FD iPSCs showed the maintenance of the molecular and biochemical characteristics and a normal karyotype. The KO cell line recapitulated the biological features observed in FD patient cells, with reduced GLA expression, lower α-Galactosidase A (α-Gal A) activity (1.5 nmol/h/mg protein), and Gb3 accumulation. The corrected cell line was generated with 75.8% efficiency and 69.6% on-target efficacy. Enzyme activity increased to 579 nmol/h/mg protein (vs. 0.78 nmol/h/mg protein in FD iPSCs), accompanied by a marked reduction in Gb3 levels. We successfully generated iPSC-CM lines, which were validated by qRT-PCR and immunofluorescence. Discussion: Cell modelling is essential for studying the pathophysiology of disease mechanisms. By retaining the characteristics of the original cells, iPSCs are a valuable biological resource for generating specific differentiated cell types affected by the disease, which would otherwise be difficult to access. This study also explored the therapeutic potential of gene editing as a promising approach to altering the course of rare diseases.eng
dc.description.sponsorshipTrabalho realizado no INSA e UP AJD- Bolsa de doutoramento FCT/MEC: SFRH/BD/118009/2016 OA- Projeto FCT/MEC: PTDC/BIM-MEC/4762/2014 Unidade de investigação FCT/MEC: UIDB/00211/2020
dc.identifier.issnNA
dc.identifier.urihttp://hdl.handle.net/10400.18/11102
dc.language.isoeng
dc.peerreviewedyes
dc.relationPTDC/BIM-MEC/4762/2014
dc.relationFabry disease: from iPS cells to Genomic Editing
dc.relationCenter for the Study of Animal Science
dc.relationUIDB/00211/2020
dc.relation.hasversionhttps://spgh.net/wp-content/uploads/2014/04/liv_resumos_spgh25-1.pdf
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectGenética Humana
dc.subjectGene Editing
dc.subjectCell models
dc.subjectiPSCs
dc.subjectFabry Disease
dc.subjectFabry disease
dc.subjectDoenças Genéticas
dc.titleGeneration of Cellular Models for Fabry Disease: Unlocking the Potential of iPSCs and Gene Editingeng
dc.typeconference object
dspace.entity.typePublication
oaire.awardNumberPTDC/BIM-MEC/4762/2014
oaire.awardNumberSFRH/BD/118009/2016
oaire.awardNumberUIDB/00211/2020
oaire.awardTitleFabry disease: from iPS cells to Genomic Editing
oaire.awardTitleCenter for the Study of Animal Science
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/3599-PPCDT/PTDC%2FBIM-MEC%2F4762%2F2014/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT//SFRH%2FBD%2F118009%2F2016/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F00211%2F2020/PT
oaire.citation.conferenceDate2025-11-20
oaire.citation.conferencePlaceCoimbra, Portugal
oaire.citation.title29th Annual Meeting of the Portuguese Society of Human Genetics (SPGH), 20-22 novembro 2025
oaire.fundingStream3599-PPCDT
oaire.fundingStream6817 - DCRRNI ID
oaire.versionhttp://purl.org/coar/version/c_970fb48d4fbd8a85
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
relation.isProjectOfPublication9452b84c-9691-47bb-82ba-75ed40cdfa04
relation.isProjectOfPublication2d0390cf-01d3-491f-9daf-ceba651b89fe
relation.isProjectOfPublication69b75eb9-6f25-4ad8-98db-6cc7e9bcdcc7
relation.isProjectOfPublication.latestForDiscovery9452b84c-9691-47bb-82ba-75ed40cdfa04

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