A new physiological medium uncovers biochemical and cellular alterations in Lesch-Nyhan disease fibroblasts
Escudero-Ferruz, Paula (Universitat Autònoma de Barcelona. Institut de Neurociències)
Ontiveros, Neus (Universitat Autònoma de Barcelona. Institut de Neurociències)
Cano-Estrada, Claudia (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
Sutcliffe, Diane J. (Emory University School of Medicine)
Jinnah, H. A. (Emory University School of Medicine)
Torres, Rosa J. (Hospital Universitario La Paz (Madrid))
López Blanco, José Manuel (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)

Data:	2024
Descripció:	20 pàg.
Resum:	Background: Lesch-Nyhan disease (LND) is a severe neurological disorder caused by the genetic deficiency of hypoxanthine-guanine phosphoribosyltransferase (HGprt), an enzyme involved in the salvage synthesis of purines. To compensate this deficiency, there is an acceleration of the de novo purine biosynthetic pathway. Most studies have failed to find any consistent abnormalities of purine nucleotides in cultured cells obtained from the patients. Recently, it has been shown that 5-aminoimidazole-4-carboxamide riboside 5ʹ-monophosphate (ZMP), an intermediate of the de novo pathway, accumulates in LND fibroblasts maintained with RPMI containing physiological levels (25 nM) of folic acid (FA), which strongly differs from FA levels of regular cell culture media (2200 nM). However, RPMI and other standard media contain non-physiological levels of many nutrients, having a great impact in cell metabolism that does not precisely recapitulate the in vivo behavior of cells. Methods: We prepared a new culture medium containing physiological levels of all nutrients, including vitamins (Plasmax-PV), to study the potential alterations of LND fibroblasts that may have been masked by the usage of non-physiological media. We quantified ZMP accumulation under different culture conditions and evaluated the activity of two known ZMP-target proteins (AMPK and ADSL), the mRNA expression of the folate carrier SLC19A1, possible mitochondrial alterations and functional consequences in LND fibroblasts. Results: LND fibroblasts maintained with Plasmax-PV show metabolic adaptations such a higher glycolytic capacity, increased expression of the folate carrier SCL19A1, and functional alterations such a decreased mitochondrial potential and reduced cell migration compared to controls. These alterations can be reverted with high levels of folic acid, suggesting that folic acid supplements might be a potential treatment for LND. Conclusions: A complete physiological cell culture medium reveals new alterations in Lesch-Nyhan disease. This work emphasizes the importance of using physiological cell culture conditions when studying a metabolic disorder.
Ajuts:	Agencia Estatal de Investigación PID2020-113124RB-C21 Agencia Estatal de Investigación PID2020-113124RB-C22 Fundació la Marató de TV3 776/C/2020
Drets:	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.
Llengua:	Anglès
Document:	Article ; recerca ; Versió publicada
Matèria:	AICAR ; Folic acid ; Lesch-Nyhan disease ; Plasmax ; Purine nucleotides ; ZMP
Publicat a:	Molecular Medicine, Vol. 30, Num. 1 (January 2024) , ISSN 1528-3658

DOI: 10.1186/s10020-023-00774-8
PMID: 38172668

20 p, 3.9 MB

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