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A microglial cell model for acyl-CoA oxidase 1 deficiency.

Abstract

Acyl-CoA oxidase 1 (ACOX1) deficiency is a rare and severe peroxisomal leukodystrophy associated with a very long-chain fatty acid (VLCFA) β-oxidation defect. This neurodegenerative disease lacks relevant cell models to further decipher the pathomechanisms in order to identify novel therapeutic targets. Since peroxisomal defects in microglia appear to be a key component of peroxisomal leukodystrophies, we targeted the Acox1 gene in the murine microglial BV-2 cell line. Using CRISPR/Cas9 gene editing, we generated an Acox1-deficient cell line and validated the allelic mutations, which lead to the absence of ACOX1 protein and enzymatic activity. The activity of catalase, the enzyme degrading H2O2, was increased, likely in response to the alteration of redox homeostasis. The mutant cell line grew more slowly than control cells without obvious morphological changes. However, ultrastructural analysis revealed an increased number of peroxisomes and mitochondria associated with size reduction of mitochondria. Changes in the distribution of lipid droplets containing neutral lipids have been observed in mutant cells; lipid analysis revealed the accumulation of saturated and monounsaturated VLCFA. Besides, expression levels of genes encoding interleukin-1 beta and 6 (IL-1β and IL-6), as well as triggering receptor expressed on myeloid cells 2 (Trem2) were found modified in the mutant cells suggesting modification of microglial polarization and phagocytosis ability. In summary, this Acox1-deficient cell line presents the main biochemical characteristics of the human disease and will serve as a promising model to further investigate the consequences of a specific microglial peroxisomal β-oxidation defect on oxidative stress, inflammation and cellular functions.

Authors+Show Affiliations

Laboratoire Bio-PeroxIL EA7270, University of Bourgogne Franche-Comté, Dijon, France.Laboratoire Bio-PeroxIL EA7270, University of Bourgogne Franche-Comté, Dijon, France; Laboratoire de Biochimie et Neurosciences, Faculté des Sciences et Techniques, University Hassan I, Settat, Morocco.Laboratoire Bio-PeroxIL EA7270, University of Bourgogne Franche-Comté, Dijon, France.Laboratoire Bio-PeroxIL EA7270, University of Bourgogne Franche-Comté, Dijon, France.Aix Marseille Univ, CNRS, INSERM, CIML, Centre d'Immunologie de Marseille-Luminy, Marseille, France.Laboratory of Clinical Chemistry, Hospital of Varese, ASST-Settelaghi, Milan, Italy.Laboratory of Medical Genetics and Neurogenetics, Foundation IRCCS Istituto Neurologico Carlo Besta, Milan, Italy.Laboratoire de Biochimie et Neurosciences, Faculté des Sciences et Techniques, University Hassan I, Settat, Morocco.Unité de Recherche en Physiologie Moléculaire (URPhyM), laboratoire de Chimie Physiologique, NARILIS (Namur Research Institute for Life Sciences), University of Namur (UNamur), Namur, Belgium.Centre des Sciences du Go슩t et de l'Alimentation, AgroSup Dijon, UMR6265/UMRA1324, CNRS, INRA, University of Bourgogne Franche-Comté, Dijon, France.Laboratoire Bio-PeroxIL EA7270, University of Bourgogne Franche-Comté, Dijon, France; INSERM, Dijon, France.Laboratoire Bio-PeroxIL EA7270, University of Bourgogne Franche-Comté, Dijon, France.Laboratoire Bio-PeroxIL EA7270, University of Bourgogne Franche-Comté, Dijon, France.Laboratoire Bio-PeroxIL EA7270, University of Bourgogne Franche-Comté, Dijon, France. Electronic address: stsavary@u-bourgogne.fr.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

30312667

Citation

Raas, Q, et al. "A Microglial Cell Model for acyl-CoA Oxidase 1 Deficiency." Biochimica Et Biophysica Acta. Molecular and Cell Biology of Lipids, vol. 1864, no. 4, 2019, pp. 567-576.
Raas Q, Saih FE, Gondcaille C, et al. A microglial cell model for acyl-CoA oxidase 1 deficiency. Biochim Biophys Acta Mol Cell Biol Lipids. 2019;1864(4):567-576.
Raas, Q., Saih, F. E., Gondcaille, C., Trompier, D., Hamon, Y., Leoni, V., ... Savary, S. (2019). A microglial cell model for acyl-CoA oxidase 1 deficiency. Biochimica Et Biophysica Acta. Molecular and Cell Biology of Lipids, 1864(4), pp. 567-576. doi:10.1016/j.bbalip.2018.10.005.
Raas Q, et al. A Microglial Cell Model for acyl-CoA Oxidase 1 Deficiency. Biochim Biophys Acta Mol Cell Biol Lipids. 2019;1864(4):567-576. PubMed PMID: 30312667.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A microglial cell model for acyl-CoA oxidase 1 deficiency. AU - Raas,Q, AU - Saih,F-E, AU - Gondcaille,C, AU - Trompier,D, AU - Hamon,Y, AU - Leoni,V, AU - Caccia,C, AU - Nasser,B, AU - Jadot,M, AU - Ménétrier,F, AU - Lizard,G, AU - Cherkaoui-Malki,M, AU - Andreoletti,P, AU - Savary,S, Y1 - 2018/10/10/ PY - 2018/06/21/received PY - 2018/10/01/revised PY - 2018/10/05/accepted PY - 2018/10/13/pubmed PY - 2019/10/23/medline PY - 2018/10/13/entrez KW - Acyl-CoA oxidase KW - Microglia KW - Peroxisome KW - VLCFA SP - 567 EP - 576 JF - Biochimica et biophysica acta. Molecular and cell biology of lipids JO - Biochim Biophys Acta Mol Cell Biol Lipids VL - 1864 IS - 4 N2 - Acyl-CoA oxidase 1 (ACOX1) deficiency is a rare and severe peroxisomal leukodystrophy associated with a very long-chain fatty acid (VLCFA) β-oxidation defect. This neurodegenerative disease lacks relevant cell models to further decipher the pathomechanisms in order to identify novel therapeutic targets. Since peroxisomal defects in microglia appear to be a key component of peroxisomal leukodystrophies, we targeted the Acox1 gene in the murine microglial BV-2 cell line. Using CRISPR/Cas9 gene editing, we generated an Acox1-deficient cell line and validated the allelic mutations, which lead to the absence of ACOX1 protein and enzymatic activity. The activity of catalase, the enzyme degrading H2O2, was increased, likely in response to the alteration of redox homeostasis. The mutant cell line grew more slowly than control cells without obvious morphological changes. However, ultrastructural analysis revealed an increased number of peroxisomes and mitochondria associated with size reduction of mitochondria. Changes in the distribution of lipid droplets containing neutral lipids have been observed in mutant cells; lipid analysis revealed the accumulation of saturated and monounsaturated VLCFA. Besides, expression levels of genes encoding interleukin-1 beta and 6 (IL-1β and IL-6), as well as triggering receptor expressed on myeloid cells 2 (Trem2) were found modified in the mutant cells suggesting modification of microglial polarization and phagocytosis ability. In summary, this Acox1-deficient cell line presents the main biochemical characteristics of the human disease and will serve as a promising model to further investigate the consequences of a specific microglial peroxisomal β-oxidation defect on oxidative stress, inflammation and cellular functions. SN - 1879-2618 UR - https://www.unboundmedicine.com/medline/citation/30312667/A_microglial_cell_model_for_acyl-CoA_oxidase_1_deficiency L2 - https://linkinghub.elsevier.com/retrieve/pii/S1388-1981(18)30324-X DB - PRIME DP - Unbound Medicine ER -