Generation of high number of healthy erythroid cells from gene-edited pyruvate kinase deficiency patient-specific induced pluripotent stem cells

Garate, Z. et al. (2015) Generation of high number of healthy erythroid cells from gene-edited pyruvate kinase deficiency patient-specific induced pluripotent stem cells. Stem Cell Reports, 5(6), pp. 1053-1066. (doi:10.1016/j.stemcr.2015.10.002) (PMID:26549847) (PMCID:PMC4682065)

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Abstract

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.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Mountford, Dr Joanne and Olivier, Dr Emmanuel
Authors: Garate, Z., Quintana-Bustamante, O., Crane, A. M., 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, T. M., Ribeiro, M. L., Sevilla, J., Prosper, F., Jin, S., Mountford, J., Guenechea, G., Gouble, A., Bueren, J. A., Davis, B. R., and Segovia, J. C.
College/School:College of Medical Veterinary and Life Sciences > Institute of Cardiovascular and Medical Sciences
Journal Name:Stem Cell Reports
Publisher:Elsevier Inc.
ISSN:2213-6711
ISSN (Online):2213-6711
Published Online:05 November 2015
Copyright Holders:Copyright © 2015 The Authors
First Published:First published in Stem Cell Reports 5(6):1053-1066
Publisher Policy:Reproduced under a Creative Commons License

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