@article{2a7a7a8f7f5c4b109b1d847a55db56f2,
title = "Generation of a High Number of Healthy Erythroid Cells from Gene-Edited Pyruvate Kinase Deficiency Patient-Specific Induced Pluripotent Stem Cells",
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.",
author = "Zita Garate and Oscar Quintana-Bustamante and Crane, {Ana M.} and Emmanuel Olivier and Laurent Poirot and Roman Galetto and Penelope Kosinski and Collin Hill and Charles Kung and Xabi Agirre and Israel Orman and Laura Cerrato and Omaira Alberquilla and Fatima Rodriguez-Fornes and Noemi Fusaki and Felix Garcia-Sanchez and Maia, {Tabita M.} and Ribeiro, {Maria L.} and Julian Sevilla and Felipe Prosper and Shengfang Jin and Joanne Mountford and Guillermo Guenechea and Agnes Gouble and Bueren, {Juan A.} and Davis, {Brian R.} and Segovia, {Jose C.}",
note = "Funding Information: The authors would like to thank Aurora de la Cal and Sergio Losada for their collaboration in the administrative work; Miguel Angel Mart{\'i}n, Jes{\'u}s Mart{\'i}nez, and Edilia de Almeida for the careful maintenance of the animals; J. Bazinet for providing logistics support for the metabolomics experiments; and Norman Feltz for reading and correcting the manuscript. This work was supported by the following grants: Direcci{\'o}n General de Investigaci{\'o}n de la Comunidad de Madrid (CellCAM; Ref S2010/BMD-2420), Fondo de Investigaciones Sanitarias, Instituto de Salud Carlos III (RETICS-RD12/0019/0023 and RD12/0019/0031), SAF2014-54885-R and SAF2012-31142 from MINECO, the Cystic Fibrosis Foundation (DAVIS12GO), and the National Heart Lung and Blood Institute (RC1HL099559). J.M. and E.O. are supported by the Scottish National Blood Transfusion Service and the Wellcome Trust Translation Award Grant 102610. Z.G. was supported by a fellowship of the Gobierno Vasco (Spain). R.G., L.P., and A.G. are employees of Cellectis. All relevant data are within the paper and its Supplemental Information files; to discuss access to materials, please contact the corresponding author. ",
year = "2015",
month = dec,
day = "8",
doi = "10.1016/j.stemcr.2015.10.002",
language = "English (US)",
volume = "5",
pages = "1053--1066",
journal = "Stem Cell Reports",
issn = "2213-6711",
publisher = "Cell Press",
number = "6",
}