A microglial cell model for acyl-CoA oxidase 1 deficiency

Q Raas, F-E Saih, C Gondcaille, D Trompier, Y Hamon, V Leoni, C Caccia, B Nasser, M Jadot, F Ménétrier, G Lizard, M Cherkaoui-Malki, P Andreoletti, S Savary

Research output: Contribution to journalArticle

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 H 2O 2, 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.

LanguageEnglish
JournalBiochimica et biophysica acta. Molecular and cell biology of lipids
Early online date2018
DOIs
Publication statusPublished - 2018
Externally publishedYes

Fingerprint

Acyl-CoA Oxidase
Cell Line
Interleukin-6
Fatty Acids
Clustered Regularly Interspaced Short Palindromic Repeats
Mitochondrial Size
Lipids
Peroxisomes
Microglia
Myeloid Cells
Interleukin-1beta
Phagocytosis
Neurodegenerative Diseases
Catalase
Oxidation-Reduction
Mitochondria
Oxidative Stress
Homeostasis
Inflammation
Gene Expression

Keywords

  • Acyl-CoA oxidase
  • Microglia
  • Peroxisome
  • VLCFA

Cite this

Raas, Q ; Saih, F-E ; Gondcaille, C ; Trompier, D ; Hamon, Y ; Leoni, V ; Caccia, C ; Nasser, B ; Jadot, M ; Ménétrier, F ; Lizard, G ; Cherkaoui-Malki, M ; Andreoletti, P ; Savary, S. / A microglial cell model for acyl-CoA oxidase 1 deficiency. In: Biochimica et biophysica acta. Molecular and cell biology of lipids. 2018.
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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 H 2O 2, 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.",
keywords = "Acyl-CoA oxidase, Microglia, Peroxisome, VLCFA",
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Raas, Q, Saih, F-E, Gondcaille, C, Trompier, D, Hamon, Y, Leoni, V, Caccia, C, Nasser, B, Jadot, M, Ménétrier, F, Lizard, G, Cherkaoui-Malki, M, Andreoletti, P & Savary, S 2018, 'A microglial cell model for acyl-CoA oxidase 1 deficiency' Biochimica et biophysica acta. Molecular and cell biology of lipids. https://doi.org/10.1016/j.bbalip.2018.10.005

A microglial cell model for acyl-CoA oxidase 1 deficiency. / Raas, Q; Saih, F-E; Gondcaille, C; Trompier, D; Hamon, Y; Leoni, V; Caccia, C; Nasser, B; Jadot, M; Ménétrier, F; Lizard, G; Cherkaoui-Malki, M; Andreoletti, P; Savary, S.

In: Biochimica et biophysica acta. Molecular and cell biology of lipids, 2018.

Research output: Contribution to journalArticle

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

N1 - Copyright © 2018. Published by Elsevier B.V.

PY - 2018

Y1 - 2018

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 H 2O 2, 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.

AB - 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 H 2O 2, 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.

KW - Acyl-CoA oxidase

KW - Microglia

KW - Peroxisome

KW - VLCFA

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DO - 10.1016/j.bbalip.2018.10.005

M3 - Article

JO - Biochimica et biophysica acta. Molecular and cell biology of lipids

T2 - Biochimica et biophysica acta. Molecular and cell biology of lipids

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SN - 1388-1981

ER -