Induced pluripotent stem cells from subjects with Lesch-Nyhan disease

• Type of Material: Article
• Authors: Diane J. Sutcliffe, Emory University; Ashok R. Dinasarapu, Emory University; Jasper E. Visser, Radboud University Nijmegen; Joery den Hoed, Emory University; Fatemeh Seifar, Emory University; Piyush Joshi, Emory University; Irene Ceballos-Picot, Hop Univ Necker; Tejas Sardar, Emory University; Ellen Hess, Emory University; Yan Sun, Emory University; Zhexing Wen, Emory University; Michael Zwick, Emory University; Hyder Jinnah, Emory University
• Language: English
• Date: 2021-04-19
• Publisher: Nature Research
• Publication Version: Final Published Version
• Copyright Statement: © 2021 Springer Nature Limited
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1038/s41598-021-87955-9
• Title of Journal or Parent Work: Scientific Reports
• Volume: 11
• Issue: 1
• Start Page: 8523
• End Page: 8523
• Grant/Funding Information: Hersenstichting Fellowship (F2014(1)-16).; Netherlands Organisation for Scientific Research (NWO) ZonMW/Veni (916.12.167) grant; Additional support came in part from the Emory Integrated Genomics Core (EIGC), which is subsidized by the Emory University School of Medicine and is one of the Emory Integrated Core Facilities, as well as the Georgia Clinical & Translational Science Alliance of the National Institutes of Health under Award Number UL1TR002378.; This work was supported by the Lesch-Nyhan Syndrome Children’s Research Foundation; Additional support was provided in part by grants to the Dystonia Coalition (U54 TR001456 and U54 NS116025), a consortium of the Rare Diseases Clinical Research Network (RDCRN) that is supported by the Office of Rare Diseases Research (ORDR) at the National Center for Advancing Clinical and Translational Studies (NCATS) in collaboration with the NINDS.; National Institute for Neurological Diseases and Stroke (NINDS) at the National Institutes of Health (NIH) through grants R56 NS102980 and R01 NS109242
• Keywords: Genotype-Phenotype Correlation; Science & Technology - Other Topics; Multidisciplinary Sciences; Hypoxanthine Phosphoribosyltransferase; Genetic mouse model; Brain; Dopamine; Purine metabolism; Science & Technology; Synthesis de-novo; HPRT deficiency; Nucleotide metabolism
• Research Categories: Chemistry, Biochemistry; Health Sciences, Epidemiology; Biology, Neuroscience; Health Sciences, Pharmacology

Abstract

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.