A microglial cell model for acyl-CoA oxidase 1 deficiency

Area of Science:

• Neuroscience
• Cell Biology
• Biochemistry

Background:

• Acyl-CoA oxidase 1 (ACOX1) deficiency causes a severe, rare peroxisomal leukodystrophy.
• A critical defect in very long-chain fatty acid (VLCFA) β-oxidation underlies this neurodegenerative disease.
• The lack of suitable cell models hinders understanding of pathomechanisms and identification of therapeutic targets.

Purpose of the Study:

• To develop a cell model for studying ACOX1 deficiency.
• To investigate the role of microglial peroxisomal defects in leukodystrophies.
• To establish a platform for identifying novel therapeutic strategies.

Main Methods:

• CRISPR/Cas9 gene editing was employed to create Acox1-deficient murine microglial BV-2 cells.
• Validation of mutations confirmed the absence of ACOX1 protein and enzymatic activity.
• Biochemical, morphological, and gene expression analyses were performed on the mutant cell line.

Main Results:

• Acox1-deficient cells showed increased catalase activity, altered redox homeostasis, and slower growth.
• Ultrastructural analysis revealed increased peroxisomes and smaller mitochondria.
• Accumulation of saturated and monounsaturated VLCFA was observed, along with modified expression of IL-1β, IL-6, and Trem2, suggesting altered microglial function.

Conclusions:

• The generated Acox1-deficient microglial cell line accurately recapitulates key biochemical features of human ACOX1 deficiency.
• This model provides a valuable tool for investigating microglial peroxisomal β-oxidation defects.
• It will facilitate further research into oxidative stress, inflammation, and cellular dysfunction in peroxisomal leukodystrophies.