title: Disruption of TCA Cycle and Glutamate Metabolism Identified by Metabolomics in an In Vitro Model of Amyotrophic Lateral Sclerosis.
abstract: This study aims to develop a cellular metabolomics model that reproduces the pathophysiological conditions found in amyotrophic lateral sclerosis in order to improve knowledge of disease physiology. We used a co-culture model combining the motor neuron-like cell line NSC-34 and the astrocyte clone C8-D1A, with each over-expressing wild-type or G93C mutant human SOD1, to examine amyotrophic lateral sclerosis (ALS) physiology. We focused on the effects of mutant human SOD1 as well as oxidative stress induced by menadione on intracellular metabolism using a metabolomics approach through gas chromatography coupled with mass spectrometry (GC-MS) analysis. Preliminary non-supervised analysis by Principal Component Analysis (PCA) revealed that cell type, genetic environment, and time of culture influenced the metabolomics profiles. Supervised analysis using orthogonal partial least squares discriminant analysis (OPLS-DA) on data from intracellular metabolomics profiles of SOD1G93Cco-cultures produced metabolites involved in glutamate metabolism and the tricarboxylic acid cycle (TCA) cycle. This study revealed the feasibility of using a metabolomics approach in a cellular model of ALS. We identified potential disruption of the TCA cycle and glutamate metabolism under oxidative stress, which is consistent with prior research in the disease. Analysis of metabolic alterations in an in vitro model is a novel approach to investigation of disease physiology.
authors.affiliation: UMR INSERM U930, Université François-Rabelais de Tours, Equipe « Neurogénétique et neurométabolomique », 37032, Tours, France. charlotte.veyrat@live.fr.|CHRU de Tours, Laboratoire de Biochimie et de biologie moléculaire, 37044, Tours, France. charlotte.veyrat@live.fr.
authors.forename: Charlotte
authors.initials: C
authors.lastname: Veyrat-Durebex
authors.affiliation: UMR INSERM U930, Université François-Rabelais de Tours, Equipe « Neurogénétique et neurométabolomique », 37032, Tours, France.|CHRU de Tours, Service de Neurologie, 37044, Tours, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Corcia
authors.affiliation: INSERM U966, 37032, Tours, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Piver
authors.affiliation: Département de Pharmacologie médicale, INSERM U1171, Université Lille Nord de France, CHRU de Lille, Lille, France.
authors.forename: David
authors.initials: D
authors.lastname: Devos
authors.affiliation: UMR INSERM U930, Université François-Rabelais de Tours, Equipe « Neurogénétique et neurométabolomique », 37032, Tours, France.
authors.forename: Audrey
authors.initials: A
authors.lastname: Dangoumau
authors.affiliation: Département de Pharmacologie médicale, INSERM U1171, Université Lille Nord de France, CHRU de Lille, Lille, France.
authors.forename: Flore
authors.initials: F
authors.lastname: Gouel
authors.affiliation: UMR INSERM U930, Université François-Rabelais de Tours, Equipe « Neurogénétique et neurométabolomique », 37032, Tours, France.|CHRU de Tours, Laboratoire de Biochimie et de biologie moléculaire, 37044, Tours, France.
authors.forename: Patrick
authors.initials: P
authors.lastname: Vourc'h
authors.affiliation: UMR INSERM U930, Université François-Rabelais de Tours, Equipe « Neurogénétique et neurométabolomique », 37032, Tours, France.|PPF-ASB, Université François Rabelais de Tours, Tours, France.
authors.forename: Patrick
authors.initials: P
authors.lastname: Emond
authors.affiliation: UMR INSERM U930, Université François-Rabelais de Tours, Equipe « Neurogénétique et neurométabolomique », 37032, Tours, France.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Laumonnier
authors.affiliation: UMR INSERM U930, Université François-Rabelais de Tours, Equipe « Neurogénétique et neurométabolomique », 37032, Tours, France.|PPF-ASB, Université François Rabelais de Tours, Tours, France.
authors.forename: Lydie
authors.initials: L
authors.lastname: Nadal-Desbarats
authors.affiliation: Northern Navajo Medical Center, Shiprock, NM, USA.
authors.forename: Paul H
authors.initials: PH
authors.lastname: Gordon
authors.affiliation: UMR INSERM U930, Université François-Rabelais de Tours, Equipe « Neurogénétique et neurométabolomique », 37032, Tours, France.|CHRU de Tours, Laboratoire de Biochimie et de biologie moléculaire, 37044, Tours, France.
authors.forename: Christian R
authors.initials: CR
authors.lastname: Andres
authors.affiliation: UMR INSERM U930, Université François-Rabelais de Tours, Equipe « Neurogénétique et neurométabolomique », 37032, Tours, France.|CHRU de Tours, Laboratoire de Biochimie et de biologie moléculaire, 37044, Tours, France.
authors.forename: Hélène
authors.initials: H
authors.lastname: Blasco
mesh_terms: D000690:Amyotrophic Lateral Sclerosis / Q000378:metabolism* / Q000473:pathology; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D002952:Citric Acid Cycle / Q000187:drug effects; D018920:Coculture Techniques; D016002:Discriminant Analysis; D008401:Gas Chromatography-Mass Spectrometry; D005971:Glutamates / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D055432:Metabolomics; D008954:Models, Biological; D018384:Oxidative Stress / Q000187:drug effects; D025341:Principal Component Analysis; D011786:Quality Control; D017382:Reactive Oxygen Species / Q000378:metabolism; D015203:Reproducibility of Results; D013482:Superoxide Dismutase / Q000378:metabolism; D013997:Time Factors; D024483:Vitamin K 3 / Q000494:pharmacology
chemical_list: D005971:Glutamates; D017382:Reactive Oxygen Species; D049452:Green Fluorescent Proteins; D024483:Vitamin K 3; D013482:Superoxide Dismutase
keywords: Astrocytes; Co-culture; GC-MS; Metabolomics; Motor neurons; Oxidative stress