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Rescue from galactose-induced death of Leigh Syndrome patient cells by pyruvate and NAD(+)

Cell models of mitochondrial complex I (CI) deficiency display activation of glycolysis to compensate for the loss in mitochondrial ATP production. This adaptation can mask other relevant deficiency-induced aberrations in cell physiology. Here we investigated the viability, mitochondrial morphofunct...

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Autores principales: Iannetti, Eligio F., Smeitink, Jan A. M., Willems, Peter H. G. M., Beyrath, Julien, Koopman, Werner J. H.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6235972/
https://www.ncbi.nlm.nih.gov/pubmed/30429455
http://dx.doi.org/10.1038/s41419-018-1179-4
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author Iannetti, Eligio F.
Smeitink, Jan A. M.
Willems, Peter H. G. M.
Beyrath, Julien
Koopman, Werner J. H.
author_facet Iannetti, Eligio F.
Smeitink, Jan A. M.
Willems, Peter H. G. M.
Beyrath, Julien
Koopman, Werner J. H.
author_sort Iannetti, Eligio F.
collection PubMed
description Cell models of mitochondrial complex I (CI) deficiency display activation of glycolysis to compensate for the loss in mitochondrial ATP production. This adaptation can mask other relevant deficiency-induced aberrations in cell physiology. Here we investigated the viability, mitochondrial morphofunction, ROS levels and ATP homeostasis of primary skin fibroblasts from Leigh Syndrome (LS) patients with isolated CI deficiency. These cell lines harbored mutations in nuclear DNA (nDNA)-encoded CI genes (NDUFS7, NDUFS8, NDUFV1) and, to prevent glycolysis upregulation, were cultured in a pyruvate-free medium in which glucose was replaced by galactose. Following optimization of the cell culture protocol, LS fibroblasts died in the galactose medium, whereas control cells did not. LS cell death was dose-dependently inhibited by pyruvate, malate, oxaloacetate, α-ketoglutarate, aspartate, and exogenous NAD(+) (eNAD), but not by lactate, succinate, α-ketobutyrate, and uridine. Pyruvate and eNAD increased the cellular NAD(+) content in galactose-treated LS cells to a different extent and co-incubation studies revealed that pyruvate-induced rescue was not primarily mediated by NAD(+). Functionally, in LS cells glucose-by-galactose replacement increased mitochondrial fragmentation and mass, depolarized the mitochondrial membrane potential (Δψ), increased H(2)DCFDA-oxidizing ROS levels, increased mitochondrial ATP generation, and reduced the total cellular ATP content. These aberrations were differentially rescued by pyruvate and eNAD, supporting the conclusion that these compounds rescue galactose-induced LS cell death via different mechanisms. These findings establish a cell-based strategy for intervention testing and enhance our understanding of CI deficiency pathophysiology.
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spelling pubmed-62359722018-11-15 Rescue from galactose-induced death of Leigh Syndrome patient cells by pyruvate and NAD(+) Iannetti, Eligio F. Smeitink, Jan A. M. Willems, Peter H. G. M. Beyrath, Julien Koopman, Werner J. H. Cell Death Dis Article Cell models of mitochondrial complex I (CI) deficiency display activation of glycolysis to compensate for the loss in mitochondrial ATP production. This adaptation can mask other relevant deficiency-induced aberrations in cell physiology. Here we investigated the viability, mitochondrial morphofunction, ROS levels and ATP homeostasis of primary skin fibroblasts from Leigh Syndrome (LS) patients with isolated CI deficiency. These cell lines harbored mutations in nuclear DNA (nDNA)-encoded CI genes (NDUFS7, NDUFS8, NDUFV1) and, to prevent glycolysis upregulation, were cultured in a pyruvate-free medium in which glucose was replaced by galactose. Following optimization of the cell culture protocol, LS fibroblasts died in the galactose medium, whereas control cells did not. LS cell death was dose-dependently inhibited by pyruvate, malate, oxaloacetate, α-ketoglutarate, aspartate, and exogenous NAD(+) (eNAD), but not by lactate, succinate, α-ketobutyrate, and uridine. Pyruvate and eNAD increased the cellular NAD(+) content in galactose-treated LS cells to a different extent and co-incubation studies revealed that pyruvate-induced rescue was not primarily mediated by NAD(+). Functionally, in LS cells glucose-by-galactose replacement increased mitochondrial fragmentation and mass, depolarized the mitochondrial membrane potential (Δψ), increased H(2)DCFDA-oxidizing ROS levels, increased mitochondrial ATP generation, and reduced the total cellular ATP content. These aberrations were differentially rescued by pyruvate and eNAD, supporting the conclusion that these compounds rescue galactose-induced LS cell death via different mechanisms. These findings establish a cell-based strategy for intervention testing and enhance our understanding of CI deficiency pathophysiology. Nature Publishing Group UK 2018-11-14 /pmc/articles/PMC6235972/ /pubmed/30429455 http://dx.doi.org/10.1038/s41419-018-1179-4 Text en © The Author(s) 2018 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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Iannetti, Eligio F.
Smeitink, Jan A. M.
Willems, Peter H. G. M.
Beyrath, Julien
Koopman, Werner J. H.
Rescue from galactose-induced death of Leigh Syndrome patient cells by pyruvate and NAD(+)
title Rescue from galactose-induced death of Leigh Syndrome patient cells by pyruvate and NAD(+)
title_full Rescue from galactose-induced death of Leigh Syndrome patient cells by pyruvate and NAD(+)
title_fullStr Rescue from galactose-induced death of Leigh Syndrome patient cells by pyruvate and NAD(+)
title_full_unstemmed Rescue from galactose-induced death of Leigh Syndrome patient cells by pyruvate and NAD(+)
title_short Rescue from galactose-induced death of Leigh Syndrome patient cells by pyruvate and NAD(+)
title_sort rescue from galactose-induced death of leigh syndrome patient cells by pyruvate and nad(+)
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6235972/
https://www.ncbi.nlm.nih.gov/pubmed/30429455
http://dx.doi.org/10.1038/s41419-018-1179-4
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