A new physiological medium uncovers biochemical and cellular alterations in Lesch-Nyhan disease fibroblasts
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 bi...
| Autores: | , , , , , , |
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| Tipo de recurso: | artículo |
| Fecha de publicación: | 2024 |
| País: | España |
| Institución: | Universitat Autònoma de Barcelona |
| Repositorio: | Dipòsit Digital de Documents de la UAB |
| Idioma: | inglés |
| OAI Identifier: | oai:ddd.uab.cat:304149 |
| Acceso en línea: | https://ddd.uab.cat/record/304149 https://dx.doi.org/urn:doi:10.1186/s10020-023-00774-8 |
| Access Level: | acceso abierto |
| Palabra clave: | AICAR Folic acid Lesch-Nyhan disease Plasmax Purine nucleotides ZMP |
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A new physiological medium uncovers biochemical and cellular alterations in Lesch-Nyhan disease fibroblastsEscudero Ferruz, Paula|||0000-0002-8778-392XOntiveros Roca, NeusCano Estrada, Claudia|||0009-0006-9177-9241Sutcliffe, Diane J.Jinnah, H. A.Torres, Rosa J.López Blanco, José Manuel|||0000-0002-4104-6262AICARFolic acidLesch-Nyhan diseasePlasmaxPurine nucleotidesZMPBackground: 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. 22024-01-0120242024-01-01Articlehttp://purl.org/coar/resource_type/c_6501VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttps://ddd.uab.cat/record/304149https://dx.doi.org/urn:doi:10.1186/s10020-023-00774-8reponame:Dipòsit Digital de Documents de la UABinstname:Universitat Autònoma de BarcelonaInglésengAgencia Estatal de Investigación https://doi.org/10.13039/501100011033 PID2020-113124RB-C21Agencia Estatal de Investigación https://doi.org/10.13039/501100011033 PID2020-113124RB-C22Fundació la Marató de TV3 https://doi.org/10.13039/100008666 776/C/2020open accesshttp://purl.org/coar/access_right/c_abf2Aquest 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.https://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:ddd.uab.cat:3041492026-06-06T12:50:31Z |
| dc.title.none.fl_str_mv |
A new physiological medium uncovers biochemical and cellular alterations in Lesch-Nyhan disease fibroblasts |
| title |
A new physiological medium uncovers biochemical and cellular alterations in Lesch-Nyhan disease fibroblasts |
| spellingShingle |
A new physiological medium uncovers biochemical and cellular alterations in Lesch-Nyhan disease fibroblasts Escudero Ferruz, Paula|||0000-0002-8778-392X AICAR Folic acid Lesch-Nyhan disease Plasmax Purine nucleotides ZMP |
| title_short |
A new physiological medium uncovers biochemical and cellular alterations in Lesch-Nyhan disease fibroblasts |
| title_full |
A new physiological medium uncovers biochemical and cellular alterations in Lesch-Nyhan disease fibroblasts |
| title_fullStr |
A new physiological medium uncovers biochemical and cellular alterations in Lesch-Nyhan disease fibroblasts |
| title_full_unstemmed |
A new physiological medium uncovers biochemical and cellular alterations in Lesch-Nyhan disease fibroblasts |
| title_sort |
A new physiological medium uncovers biochemical and cellular alterations in Lesch-Nyhan disease fibroblasts |
| dc.creator.none.fl_str_mv |
Escudero Ferruz, Paula|||0000-0002-8778-392X Ontiveros Roca, Neus Cano Estrada, Claudia|||0009-0006-9177-9241 Sutcliffe, Diane J. Jinnah, H. A. Torres, Rosa J. López Blanco, José Manuel|||0000-0002-4104-6262 |
| author |
Escudero Ferruz, Paula|||0000-0002-8778-392X |
| author_facet |
Escudero Ferruz, Paula|||0000-0002-8778-392X Ontiveros Roca, Neus Cano Estrada, Claudia|||0009-0006-9177-9241 Sutcliffe, Diane J. Jinnah, H. A. Torres, Rosa J. López Blanco, José Manuel|||0000-0002-4104-6262 |
| author_role |
author |
| author2 |
Ontiveros Roca, Neus Cano Estrada, Claudia|||0009-0006-9177-9241 Sutcliffe, Diane J. Jinnah, H. A. Torres, Rosa J. López Blanco, José Manuel|||0000-0002-4104-6262 |
| author2_role |
author author author author author author |
| dc.subject.none.fl_str_mv |
AICAR Folic acid Lesch-Nyhan disease Plasmax Purine nucleotides ZMP |
| topic |
AICAR Folic acid Lesch-Nyhan disease Plasmax Purine nucleotides ZMP |
| description |
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. |
| publishDate |
2024 |
| dc.date.none.fl_str_mv |
2 2024-01-01 2024 2024-01-01 |
| dc.type.none.fl_str_mv |
Article http://purl.org/coar/resource_type/c_6501 VoR http://purl.org/coar/version/c_970fb48d4fbd8a85 |
| dc.type.openaire.fl_str_mv |
info:eu-repo/semantics/article |
| format |
article |
| dc.identifier.none.fl_str_mv |
https://ddd.uab.cat/record/304149 https://dx.doi.org/urn:doi:10.1186/s10020-023-00774-8 |
| url |
https://ddd.uab.cat/record/304149 https://dx.doi.org/urn:doi:10.1186/s10020-023-00774-8 |
| dc.language.none.fl_str_mv |
Inglés eng |
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Inglés |
| language |
eng |
| dc.relation.none.fl_str_mv |
Agencia Estatal de Investigación https://doi.org/10.13039/501100011033 PID2020-113124RB-C21 Agencia Estatal de Investigación https://doi.org/10.13039/501100011033 PID2020-113124RB-C22 Fundació la Marató de TV3 https://doi.org/10.13039/100008666 776/C/2020 |
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open access http://purl.org/coar/access_right/c_abf2 https://creativecommons.org/licenses/by/4.0/ |
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info:eu-repo/semantics/openAccess |
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open access http://purl.org/coar/access_right/c_abf2 https://creativecommons.org/licenses/by/4.0/ |
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openAccess |
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