Generation of a High Number of Healthy Erythroid Cells from Gene-Edited Pyruvate Kinase Deficiency Patient-Specific Induced Pluripotent Stem Cells

Detalhes bibliográficos
Autor(a) principal: Garate, Z
Data de Publicação: 2015
Outros Autores: Quintana-Bustamante, O, Crane, AM, Olivier, E, Poirot, L, Galetto, R, Kosinski, P, Hill, C, Kung, C, Agirre, X, Orman, I, Cerrato, L, Alberquilla, O, Rodriguez-Fornes, F, Fusaki, N, Garcia-Sanchez, F, Maia, TM, Ribeiro, ML, Sevilla, J, Prosper, F, Jin, S, Mountford, J, Guenechea, G, Gouble, A, Bueren, JA, Davis, BR, Segovia, JC
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10400.4/2083
Resumo: Pyruvate kinase deficiency (PKD) is a rare erythroid metabolic disease caused by mutations in the PKLR gene. Erythrocytes from PKD patients show an energetic imbalance causing chronic non-spherocytic hemolytic anemia, as pyruvate kinase defects impair ATP production in erythrocytes. We generated PKD induced pluripotent stem cells (PKDiPSCs) from peripheral blood mononuclear cells (PB-MNCs) of PKD patients by non-integrative Sendai viral vectors. PKDiPSCs were gene edited to integrate a partial codon-optimized R-type pyruvate kinase cDNA in the second intron of the PKLR gene by TALEN-mediated homologous recombination (HR). Notably, we found allele specificity of HR led by the presence of a single-nucleotide polymorphism. High numbers of erythroid cells derived from gene-edited PKDiPSCs showed correction of the energetic imbalance, providing an approach to correct metabolic erythroid diseases and demonstrating the practicality of this approach to generate the large cell numbers required for comprehensive biochemical and metabolic erythroid analyses.
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spelling Generation of a High Number of Healthy Erythroid Cells from Gene-Edited Pyruvate Kinase Deficiency Patient-Specific Induced Pluripotent Stem CellsAnemia Hemolítica Congénita não EsferocíticaCélulas EritroidesCélulas-Tronco Pluripotentes InduzidasPiruvato QuinaseErros Inatos do Metabolismo dos PiruvatosPyruvate kinase deficiency (PKD) is a rare erythroid metabolic disease caused by mutations in the PKLR gene. Erythrocytes from PKD patients show an energetic imbalance causing chronic non-spherocytic hemolytic anemia, as pyruvate kinase defects impair ATP production in erythrocytes. We generated PKD induced pluripotent stem cells (PKDiPSCs) from peripheral blood mononuclear cells (PB-MNCs) of PKD patients by non-integrative Sendai viral vectors. PKDiPSCs were gene edited to integrate a partial codon-optimized R-type pyruvate kinase cDNA in the second intron of the PKLR gene by TALEN-mediated homologous recombination (HR). Notably, we found allele specificity of HR led by the presence of a single-nucleotide polymorphism. High numbers of erythroid cells derived from gene-edited PKDiPSCs showed correction of the energetic imbalance, providing an approach to correct metabolic erythroid diseases and demonstrating the practicality of this approach to generate the large cell numbers required for comprehensive biochemical and metabolic erythroid analyses.RIHUCGarate, ZQuintana-Bustamante, OCrane, AMOlivier, EPoirot, LGaletto, RKosinski, PHill, CKung, CAgirre, XOrman, ICerrato, LAlberquilla, ORodriguez-Fornes, FFusaki, NGarcia-Sanchez, FMaia, TMRibeiro, MLSevilla, JProsper, FJin, SMountford, JGuenechea, GGouble, ABueren, JADavis, BRSegovia, JC2017-08-29T15:21:32Z2015-12-082015-12-08T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.4/2083engStem Cell Reports. 2015 Dec 8;5(6):1053-66.10.1016/j.stemcr.2015.10.002info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-07-11T14:23:23Zoai:rihuc.huc.min-saude.pt:10400.4/2083Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T18:04:33.322636Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Generation of a High Number of Healthy Erythroid Cells from Gene-Edited Pyruvate Kinase Deficiency Patient-Specific Induced Pluripotent Stem Cells
title Generation of a High Number of Healthy Erythroid Cells from Gene-Edited Pyruvate Kinase Deficiency Patient-Specific Induced Pluripotent Stem Cells
spellingShingle Generation of a High Number of Healthy Erythroid Cells from Gene-Edited Pyruvate Kinase Deficiency Patient-Specific Induced Pluripotent Stem Cells
Garate, Z
Anemia Hemolítica Congénita não Esferocítica
Células Eritroides
Células-Tronco Pluripotentes Induzidas
Piruvato Quinase
Erros Inatos do Metabolismo dos Piruvatos
title_short Generation of a High Number of Healthy Erythroid Cells from Gene-Edited Pyruvate Kinase Deficiency Patient-Specific Induced Pluripotent Stem Cells
title_full Generation of a High Number of Healthy Erythroid Cells from Gene-Edited Pyruvate Kinase Deficiency Patient-Specific Induced Pluripotent Stem Cells
title_fullStr Generation of a High Number of Healthy Erythroid Cells from Gene-Edited Pyruvate Kinase Deficiency Patient-Specific Induced Pluripotent Stem Cells
title_full_unstemmed Generation of a High Number of Healthy Erythroid Cells from Gene-Edited Pyruvate Kinase Deficiency Patient-Specific Induced Pluripotent Stem Cells
title_sort Generation of a High Number of Healthy Erythroid Cells from Gene-Edited Pyruvate Kinase Deficiency Patient-Specific Induced Pluripotent Stem Cells
author Garate, Z
author_facet Garate, Z
Quintana-Bustamante, O
Crane, AM
Olivier, E
Poirot, L
Galetto, R
Kosinski, P
Hill, C
Kung, C
Agirre, X
Orman, I
Cerrato, L
Alberquilla, O
Rodriguez-Fornes, F
Fusaki, N
Garcia-Sanchez, F
Maia, TM
Ribeiro, ML
Sevilla, J
Prosper, F
Jin, S
Mountford, J
Guenechea, G
Gouble, A
Bueren, JA
Davis, BR
Segovia, JC
author_role author
author2 Quintana-Bustamante, O
Crane, AM
Olivier, E
Poirot, L
Galetto, R
Kosinski, P
Hill, C
Kung, C
Agirre, X
Orman, I
Cerrato, L
Alberquilla, O
Rodriguez-Fornes, F
Fusaki, N
Garcia-Sanchez, F
Maia, TM
Ribeiro, ML
Sevilla, J
Prosper, F
Jin, S
Mountford, J
Guenechea, G
Gouble, A
Bueren, JA
Davis, BR
Segovia, JC
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv RIHUC
dc.contributor.author.fl_str_mv Garate, Z
Quintana-Bustamante, O
Crane, AM
Olivier, E
Poirot, L
Galetto, R
Kosinski, P
Hill, C
Kung, C
Agirre, X
Orman, I
Cerrato, L
Alberquilla, O
Rodriguez-Fornes, F
Fusaki, N
Garcia-Sanchez, F
Maia, TM
Ribeiro, ML
Sevilla, J
Prosper, F
Jin, S
Mountford, J
Guenechea, G
Gouble, A
Bueren, JA
Davis, BR
Segovia, JC
dc.subject.por.fl_str_mv Anemia Hemolítica Congénita não Esferocítica
Células Eritroides
Células-Tronco Pluripotentes Induzidas
Piruvato Quinase
Erros Inatos do Metabolismo dos Piruvatos
topic Anemia Hemolítica Congénita não Esferocítica
Células Eritroides
Células-Tronco Pluripotentes Induzidas
Piruvato Quinase
Erros Inatos do Metabolismo dos Piruvatos
description Pyruvate kinase deficiency (PKD) is a rare erythroid metabolic disease caused by mutations in the PKLR gene. Erythrocytes from PKD patients show an energetic imbalance causing chronic non-spherocytic hemolytic anemia, as pyruvate kinase defects impair ATP production in erythrocytes. We generated PKD induced pluripotent stem cells (PKDiPSCs) from peripheral blood mononuclear cells (PB-MNCs) of PKD patients by non-integrative Sendai viral vectors. PKDiPSCs were gene edited to integrate a partial codon-optimized R-type pyruvate kinase cDNA in the second intron of the PKLR gene by TALEN-mediated homologous recombination (HR). Notably, we found allele specificity of HR led by the presence of a single-nucleotide polymorphism. High numbers of erythroid cells derived from gene-edited PKDiPSCs showed correction of the energetic imbalance, providing an approach to correct metabolic erythroid diseases and demonstrating the practicality of this approach to generate the large cell numbers required for comprehensive biochemical and metabolic erythroid analyses.
publishDate 2015
dc.date.none.fl_str_mv 2015-12-08
2015-12-08T00:00:00Z
2017-08-29T15:21:32Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10400.4/2083
url http://hdl.handle.net/10400.4/2083
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Stem Cell Reports. 2015 Dec 8;5(6):1053-66.
10.1016/j.stemcr.2015.10.002
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
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repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
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