Gene expression analysis of HPRT-deficient cells maintained with physiological levels of folic acid
Torres, Rosa (Hospital Universitario La Paz (Madrid). Departamento de Bioquímica)
Valentines-Casas, Gerard (Universitat Autònoma de Barcelona. Institut de Neurociències)
Cano-Estrada, Claudia (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
Ontiveros, Neus (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
López Blanco, José Manuel (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)

Date:	2025
Abstract:	Lesch-Nyhan disease (LND) is associated with a complete deficiency of hypoxanthine-guanine phosphoribosyltransferase (HPRT) activity due to mutations in the HPRT1 gene. Although the physiopathology of LND-related neurological manifestations remains unknown, a defective neuronal developmental process is the most widely accepted hypothesis. We generated an HPRT-deficient line from the pluripotent human embryonic cell line NT2/D1 by CRISPR-Cas9 and induced its differentiation along neuroectodermal lineages by retinoic acid treatment. As levels of folic acid in the culture media may affect results in LND models, we employed physiological levels of folate. The effect of HPRT deficiency on neural development-related gene expression was evaluated using two methodological approaches: a directed qPCR array of genes related to neuronal differentiation, and global gene expression by RNAseq. HPRT-deficient pluripotent cells presented altered expression of genes related to pluripotency in human embryonic stem cells, such as DPPA3 and CFAP95, along with genes of the homeobox gene family. HPRT-deficient pluripotent cells were able to differentiate along neuro-ectodermal lineages but presented consistent dysregulation of several genes from the homeobox gene family, including EN1 and LMX1A. GO enrichment analysis of up- and downregulated genes in HPRT-deficient cells showed that the most significant biological processes affected are related to development and nervous system development.
Grants:	Agencia Estatal de Investigación PID2020-113124RB-C21 Agencia Estatal de Investigación PID2020-113124RB-C22 Fundació la Marató de TV3 https://doi.org/10.13039/100008666 776/C/2020
Note:	This work was supported by the Centre for Biomedical Network Research on Rare Diseases (CIBERER), ISCIII, Spain.
Rights:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Language:	Anglès
Document:	Article ; recerca ; Versió publicada
Subject:	Lesch-Nyhan ; HPRT ; Desenvolupament del sistema nerviós ; RNAseq ; Purina
Published in:	Cells, Vol. 14, issue 14 (July 2025) , art. 1105, ISSN 2073-4409

DOI: 10.3390/cells14141105
PMID: 40710358

26 p, 1.1 MB

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Research literature > UAB research groups literature > Research Centres and Groups (research output) > Health sciences and biosciences > Institut de Neurociències (INc)
Articles > Research articles
Articles > Published articles