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A High-Throughput Comet Assay Approach for Assessing Cellular DNA Damage
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Oxidative DNA Damage and Repair Dynamics in Multiple Sclerosis: Insights from Comet Assay Kinetics, Base Excision
Beata Filipek1,2, Anna Macieja1, Aleksandra Binda3
1Department of Microbiology and Pharmaceutical Biochemistry, Medical University of Lodz, Mazowiecka 5, 92-215 Lodz, Poland.
Biomolecules
|June 26, 2025
Summary
Multiple sclerosis patients show impaired oxidative DNA repair due to defects in base excision repair (BER) pathways, affecting disease progression. This suggests BER pathways are potential therapeutic targets for multiple sclerosis.
Area of Science:
- Neuroscience
- Genetics
- Molecular Biology
Background:
- Multiple sclerosis (MS) is a neuroinflammatory disease.
- Oxidative stress and DNA damage are implicated in MS progression.
- Base excision repair (BER) pathways are crucial for DNA repair.
Purpose of the Study:
- To investigate if defects in BER pathways contribute to MS.
- To analyze DNA repair kinetics, gene expression, and genetic variations in MS patients.
Main Methods:
- Functional DNA repair assays (comet assay) on peripheral blood mononuclear cells from 70 MS patients and 61 controls.
- Oxidative stress induction using tert-butyl hydroperoxide (TBH).
- Quantitative mRNA expression analysis of nine key BER genes and genotyping of relevant single nucleotide polymorphisms (SNPs).
Main Results:
- MS patients exhibited significantly higher TBH-induced DNA lesions and distinct DNA repair kinetics (p < 0.001).
- Reduced mRNA expression of BER genes MBD4 and NTHL1 was observed in MS patients (p < 0.0001).
- Specific BER-related SNPs (rs3087404, rs4135054, rs1052133) were associated with ineffective DNA repair.
Conclusions:
- Impaired oxidative DNA repair is present in multiple sclerosis.
- Deficits in BER pathway kinetics, gene expression, and polymorphisms contribute to MS pathology.
- DNA repair pathways represent potential therapeutic and prognostic targets for MS.
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