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Induced pluripotent stem cells from subjects with Lesch-Nyhan disease

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 co...

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Autores principales: Sutcliffe, Diane J., Dinasarapu, Ashok R., Visser, Jasper E., Hoed, Joery den, Seifar, Fatemeh, Joshi, Piyush, Ceballos-Picot, Irene, Sardar, Tejas, Hess, Ellen J., Sun, Yan V., Wen, Zhexing, Zwick, Michael E., Jinnah, H. A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8055678/
https://www.ncbi.nlm.nih.gov/pubmed/33875724
http://dx.doi.org/10.1038/s41598-021-87955-9
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author Sutcliffe, Diane J.
Dinasarapu, Ashok R.
Visser, Jasper E.
Hoed, Joery den
Seifar, Fatemeh
Joshi, Piyush
Ceballos-Picot, Irene
Sardar, Tejas
Hess, Ellen J.
Sun, Yan V.
Wen, Zhexing
Zwick, Michael E.
Jinnah, H. A.
author_facet Sutcliffe, Diane J.
Dinasarapu, Ashok R.
Visser, Jasper E.
Hoed, Joery den
Seifar, Fatemeh
Joshi, Piyush
Ceballos-Picot, Irene
Sardar, Tejas
Hess, Ellen J.
Sun, Yan V.
Wen, Zhexing
Zwick, Michael E.
Jinnah, H. A.
author_sort Sutcliffe, Diane J.
collection PubMed
description 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.
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spelling pubmed-80556782021-04-22 Induced pluripotent stem cells from subjects with Lesch-Nyhan disease Sutcliffe, Diane J. Dinasarapu, Ashok R. Visser, Jasper E. Hoed, Joery den Seifar, Fatemeh Joshi, Piyush Ceballos-Picot, Irene Sardar, Tejas Hess, Ellen J. Sun, Yan V. Wen, Zhexing Zwick, Michael E. Jinnah, H. A. Sci Rep Article 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. Nature Publishing Group UK 2021-04-19 /pmc/articles/PMC8055678/ /pubmed/33875724 http://dx.doi.org/10.1038/s41598-021-87955-9 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Sutcliffe, Diane J.
Dinasarapu, Ashok R.
Visser, Jasper E.
Hoed, Joery den
Seifar, Fatemeh
Joshi, Piyush
Ceballos-Picot, Irene
Sardar, Tejas
Hess, Ellen J.
Sun, Yan V.
Wen, Zhexing
Zwick, Michael E.
Jinnah, H. A.
Induced pluripotent stem cells from subjects with Lesch-Nyhan disease
title Induced pluripotent stem cells from subjects with Lesch-Nyhan disease
title_full Induced pluripotent stem cells from subjects with Lesch-Nyhan disease
title_fullStr Induced pluripotent stem cells from subjects with Lesch-Nyhan disease
title_full_unstemmed Induced pluripotent stem cells from subjects with Lesch-Nyhan disease
title_short Induced pluripotent stem cells from subjects with Lesch-Nyhan disease
title_sort induced pluripotent stem cells from subjects with lesch-nyhan disease
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8055678/
https://www.ncbi.nlm.nih.gov/pubmed/33875724
http://dx.doi.org/10.1038/s41598-021-87955-9
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