Induced pluripotent stem cells from subjects with Lesch-Nyhan disease.

Autor:	Sutcliffe DJ; Department of Neurology, Emory University School of Medicine, 101 Woodruff Circle, 6305 Woodruff Memorial Building, Atlanta, GA, 30322, USA., Dinasarapu AR; Department of Human Genetics, Emory University School of Medicine, Atlanta, GA, 30322, USA., Visser JE; Department of Neurology, Cognition and Behavior, Donders Institute for Brain, Radboud University Medical Center, Nijmegen, The Netherlands.; Department of Neurology, Amphia Hospital, Breda, The Netherlands., Hoed JD; Department of Neurology, Emory University School of Medicine, 101 Woodruff Circle, 6305 Woodruff Memorial Building, Atlanta, GA, 30322, USA., Seifar F; Department of Neurology, Emory University School of Medicine, 101 Woodruff Circle, 6305 Woodruff Memorial Building, Atlanta, GA, 30322, USA.; Neurosciences Graduate Program, Graduate Division of Biological and Biomedical Sciences, Laney Graduate School, Emory University, Atlanta, 30322, USA., Joshi P; Department of Neurology, Emory University School of Medicine, 101 Woodruff Circle, 6305 Woodruff Memorial Building, Atlanta, GA, 30322, USA., Ceballos-Picot I; Laboratoire de Biochimie Métabolomique Et Protéomique, Hôpital Universitaire Necker, Paris, France., Sardar T; Department of Neurology, Emory University School of Medicine, 101 Woodruff Circle, 6305 Woodruff Memorial Building, Atlanta, GA, 30322, USA., Hess EJ; Department of Neurology, Emory University School of Medicine, 101 Woodruff Circle, 6305 Woodruff Memorial Building, Atlanta, GA, 30322, USA.; Neurosciences Graduate Program, Graduate Division of Biological and Biomedical Sciences, Laney Graduate School, Emory University, Atlanta, 30322, USA.; Department of Pharmacology and Chemical Biology, Emory University School of Medicine, Atlanta, GA, 30322, USA., Sun YV; Department of Epidemiology, Emory University Rollins School of Public Health, Atlanta, GA. 30322, USA., Wen Z; Department of Neurology, Emory University School of Medicine, 101 Woodruff Circle, 6305 Woodruff Memorial Building, Atlanta, GA, 30322, USA.; Department of Psychiatry & Behavioral Sciences, Emory University School of Medicine, Atlanta, GA, 30322, USA.; Department of Cell Biology, Emory University School of Medicine, Atlanta, GA, 30322, USA., Zwick ME; Department of Human Genetics, Emory University School of Medicine, Atlanta, GA, 30322, USA.; Department of Pediatrics, Emory University School of Medicine, Atlanta, GA, 30322, USA., Jinnah HA; Department of Neurology, Emory University School of Medicine, 101 Woodruff Circle, 6305 Woodruff Memorial Building, Atlanta, GA, 30322, USA. hjinnah@emory.edu.; Department of Human Genetics, Emory University School of Medicine, Atlanta, GA, 30322, USA. hjinnah@emory.edu.; Neurosciences Graduate Program, Graduate Division of Biological and Biomedical Sciences, Laney Graduate School, Emory University, Atlanta, 30322, USA. hjinnah@emory.edu.; Department of Pediatrics, Emory University School of Medicine, Atlanta, GA, 30322, USA. hjinnah@emory.edu.
Jazyk:	angličtina
Zdroj:	Scientific reports [Sci Rep] 2021 Apr 19; Vol. 11 (1), pp. 8523. Date of Electronic Publication: 2021 Apr 19.
DOI:	10.1038/s41598-021-87955-9
Abstrakt:	Lesch-Nyhan disease (LND) is an inherited disorder caused by pathogenic variants in the HPRT1 gene, which encodes the purine recycling enzyme hypoxanthine-guanine phosphoribosyltransferase (HGprt). We generated 6 induced pluripotent stem cell (iPSC) lines from 3 individuals with LND, along with 6 control lines from 3 normal individuals. All 12 lines had the characteristics of pluripotent stem cells, as assessed by immunostaining for pluripotency markers, expression of pluripotency genes, and differentiation into the 3 primary germ cell layers. Gene expression profiling with RNAseq demonstrated significant heterogeneity among the lines. Despite this heterogeneity, several anticipated abnormalities were readily detectable across all LND lines, including reduced HPRT1 mRNA. Several unexpected abnormalities were also consistently detectable across the LND lines, including decreases in FAR2P1 and increases in RNF39. Shotgun proteomics also demonstrated several expected abnormalities in the LND lines, such as absence of HGprt protein. The proteomics study also revealed several unexpected abnormalities across the LND lines, including increases in GNAO1 decreases in NSE4A. There was a good but partial correlation between abnormalities revealed by the RNAseq and proteomics methods. Finally, functional studies demonstrated LND lines had no HGprt enzyme activity and resistance to the toxic pro-drug 6-thioguanine. Intracellular purines in the LND lines were normal, but they did not recycle hypoxanthine. These cells provide a novel resource to reveal insights into the relevance of heterogeneity among iPSC lines and applications for modeling LND.
Databáze:	MEDLINE
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