@article{2c0e59b926c34a19bd2476209b26dbe7,
title = "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 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, Cell Line, Acyl-CoA Oxidase/deficiency, Fatty Acids/metabolism, Cell Proliferation, Oxidative Stress, Hydrogen Peroxide/metabolism, Neurodegenerative Diseases/genetics, Microglia/cytology, Gene Editing, Animals, Models, Biological, CRISPR-Cas Systems, Fatty Acids, Unsaturated/metabolism, Mice, Mutation",
author = "Q Raas and F-E Saih and C Gondcaille and D Trompier and Y Hamon and V Leoni and C Caccia and B Nasser and M Jadot and F M{\'e}n{\'e}trier and G Lizard and M Cherkaoui-Malki and P Andreoletti and S Savary",
note = "Funding Information: This work was supported by the Minist{\`e}re de l'Education Nationale, de l'Enseignement Superieur et de la Recherche (France) and by institutional grants from Inserm , CNRS and Aix-Marseille University to the CIML. Quentin Raas received a fellowship from ARSEP (Association for Research on Multiple Sclerosis). Fatima Saih received a fellowship “Action Int{\'e}gr{\'e}e of the Comit{\'e} Mixte Inter-universitaire Franco-Marocain” (CMIFM, AIMA/14/310, Campus France N° 30293PA) from the PHC Toubkal program, Minist{\`e}re des Affaires Etrang{\`e}res. We thank the DImaCell platform for technical assistance in TEM (Microscopy Centre INRA/UB) and the cytometry platform of Dijon for cell sorting (INSERM/UB). We acknowledge the PICSL imaging facility of the CIML (ImagImm), member of the national infrastructure France-BioImaging supported by the French National Research Agency ( ANR-10-INBS-04 ). We thank Myriam Baes for providing us the Cre expression vector as well as Hai Tao He and Didier Marguet for hosting Quentin Raas for a few months at CIML. Funding Information: This work was supported by the Minist{\`e}re de l'Education Nationale, de l'Enseignement Superieur et de la Recherche (France) and by institutional grants from Inserm, CNRS and Aix-Marseille University to the CIML. Quentin Raas received a fellowship from ARSEP (Association for Research on Multiple Sclerosis). Fatima Saih received a fellowship “Action Int{\'e}gr{\'e}e of the Comit{\'e} Mixte Inter-universitaire Franco-Marocain” (CMIFM, AIMA/14/310, Campus France N° 30293PA) from the PHC Toubkal program, Minist{\`e}re des Affaires Etrang{\`e}res. We thank the DImaCell platform for technical assistance in TEM (Microscopy Centre INRA/UB) and the cytometry platform of Dijon for cell sorting (INSERM/UB). We acknowledge the PICSL imaging facility of the CIML (ImagImm), member of the national infrastructure France-BioImaging supported by the French National Research Agency (ANR-10-INBS-04). We thank Myriam Baes for providing us the Cre expression vector as well as Hai Tao He and Didier Marguet for hosting Quentin Raas for a few months at CIML. Publisher Copyright: {\textcopyright} 2018 Elsevier B.V.",
year = "2019",
month = apr,
doi = "10.1016/j.bbalip.2018.10.005",
language = "English",
volume = "1864",
pages = "567--576",
journal = "Biochimica et biophysica acta. Molecular and cell biology of lipids",
issn = "1388-1981",
publisher = "Elsevier",
number = "4",
}