The Association between Inefficient Repair of DNA Double-Strand Breaks and Common Polymorphisms of the HRR and NHEJ Repair Genes in Patients with Rheumatoid Arthritis
Rheumatoid arthritis (RA) is an autoimmune disease characterized by chronic inflammation affecting up to 2.0% of adults around the world. The molecular background of RA has not yet been fully elucidated, but RA is classified as a disease in which the genetic background is one of the most significant...
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MDPI AG
2024-02-01
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| Series: | International Journal of Molecular Sciences |
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| author | Grzegorz Galita Joanna Sarnik Olga Brzezinska Tomasz Budlewski Marta Poplawska Sebastian Sakowski Grzegorz Dudek Ireneusz Majsterek Joanna Makowska Tomasz Poplawski |
| author_facet | Grzegorz Galita Joanna Sarnik Olga Brzezinska Tomasz Budlewski Marta Poplawska Sebastian Sakowski Grzegorz Dudek Ireneusz Majsterek Joanna Makowska Tomasz Poplawski |
| author_sort | Grzegorz Galita |
| collection | DOAJ |
| description | Rheumatoid arthritis (RA) is an autoimmune disease characterized by chronic inflammation affecting up to 2.0% of adults around the world. The molecular background of RA has not yet been fully elucidated, but RA is classified as a disease in which the genetic background is one of the most significant risk factors. One hallmark of RA is impaired DNA repair observed in patient-derived peripheral blood mononuclear cells (PBMCs). The aim of this study was to correlate the phenotype defined as the efficiency of DNA double-strand break (DSB) repair with the genotype limited to a single-nucleotide polymorphism (SNP) of DSB repair genes. We also analyzed the expression level of key DSB repair genes. The study population contained 45 RA patients and 45 healthy controls. We used a comet assay to study DSB repair after in vitro exposure to bleomycin in PBMCs from patients with rheumatoid arthritis. TaqMan SNP Genotyping Assays were used to determine the distribution of SNPs and the Taq Man gene expression assay was used to assess the RNA expression of DSB repair-related genes. PBMCs from patients with RA had significantly lower bleomycin-induced DNA lesion repair efficiency and we identified more subjects with inefficient DNA repair in RA compared with the control (84.5% vs. 24.4%; OR 41.4, 95% CI, 4.8–355.01). Furthermore, SNPs located within the RAD50 gene (rs1801321 and rs1801320) increased the OR to 53.5 (95% CI, 4.7–613.21) while rs963917 and rs3784099 (RAD51B) to 73.4 (95% CI, 5.3–1011.05). These results were confirmed by decision tree (DT) analysis (accuracy 0.84; precision 0.87, and specificity 0.86). We also found elevated expression of RAD51B, BRCA1, and BRCA2 in PBMCs isolated from RA patients. The findings indicated that impaired DSB repair in RA may be related to genetic variations in DSB repair genes as well as their expression levels. However, the mechanism of this relation, and whether it is direct or indirect, needs to be elucidated. |
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| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-04-25T00:29:15Z |
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| spelling | doaj.art-a9c9ee6f333f4796b520fc672da98ccd2024-03-12T16:45:40ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672024-02-01255261910.3390/ijms25052619The Association between Inefficient Repair of DNA Double-Strand Breaks and Common Polymorphisms of the HRR and NHEJ Repair Genes in Patients with Rheumatoid ArthritisGrzegorz Galita0Joanna Sarnik1Olga Brzezinska2Tomasz Budlewski3Marta Poplawska4Sebastian Sakowski5Grzegorz Dudek6Ireneusz Majsterek7Joanna Makowska8Tomasz Poplawski9Department of Clinical Chemistry and Biochemistry, Medical University of Lodz, 92-215 Lodz, PolandDepartment of Rheumatology, Medical University of Lodz, 92-115 Lodz, PolandDepartment of Rheumatology, Medical University of Lodz, 92-115 Lodz, PolandDepartment of Rheumatology, Medical University of Lodz, 92-115 Lodz, PolandBiobank, Department of Immunology and Allergy, Medical University of Lodz, 92-213 Lodz, PolandFaculty of Mathematics and Computer Science, University of Lodz, 90-238 Lodz, PolandFaculty of Mathematics and Computer Science, University of Lodz, 90-238 Lodz, PolandDepartment of Clinical Chemistry and Biochemistry, Medical University of Lodz, 92-215 Lodz, PolandDepartment of Rheumatology, Medical University of Lodz, 92-115 Lodz, PolandDepartment of Pharmaceutical Microbiology and Biochemistry, Medical University of Lodz, 92-215 Lodz, PolandRheumatoid arthritis (RA) is an autoimmune disease characterized by chronic inflammation affecting up to 2.0% of adults around the world. The molecular background of RA has not yet been fully elucidated, but RA is classified as a disease in which the genetic background is one of the most significant risk factors. One hallmark of RA is impaired DNA repair observed in patient-derived peripheral blood mononuclear cells (PBMCs). The aim of this study was to correlate the phenotype defined as the efficiency of DNA double-strand break (DSB) repair with the genotype limited to a single-nucleotide polymorphism (SNP) of DSB repair genes. We also analyzed the expression level of key DSB repair genes. The study population contained 45 RA patients and 45 healthy controls. We used a comet assay to study DSB repair after in vitro exposure to bleomycin in PBMCs from patients with rheumatoid arthritis. TaqMan SNP Genotyping Assays were used to determine the distribution of SNPs and the Taq Man gene expression assay was used to assess the RNA expression of DSB repair-related genes. PBMCs from patients with RA had significantly lower bleomycin-induced DNA lesion repair efficiency and we identified more subjects with inefficient DNA repair in RA compared with the control (84.5% vs. 24.4%; OR 41.4, 95% CI, 4.8–355.01). Furthermore, SNPs located within the RAD50 gene (rs1801321 and rs1801320) increased the OR to 53.5 (95% CI, 4.7–613.21) while rs963917 and rs3784099 (RAD51B) to 73.4 (95% CI, 5.3–1011.05). These results were confirmed by decision tree (DT) analysis (accuracy 0.84; precision 0.87, and specificity 0.86). We also found elevated expression of RAD51B, BRCA1, and BRCA2 in PBMCs isolated from RA patients. The findings indicated that impaired DSB repair in RA may be related to genetic variations in DSB repair genes as well as their expression levels. However, the mechanism of this relation, and whether it is direct or indirect, needs to be elucidated.https://www.mdpi.com/1422-0067/25/5/2619rheumatoid arthritiscomet assayDNA double-strand breakDNA repairsingle-nucleotide polymorphismRAD50 |
| spellingShingle | Grzegorz Galita Joanna Sarnik Olga Brzezinska Tomasz Budlewski Marta Poplawska Sebastian Sakowski Grzegorz Dudek Ireneusz Majsterek Joanna Makowska Tomasz Poplawski The Association between Inefficient Repair of DNA Double-Strand Breaks and Common Polymorphisms of the HRR and NHEJ Repair Genes in Patients with Rheumatoid Arthritis International Journal of Molecular Sciences rheumatoid arthritis comet assay DNA double-strand break DNA repair single-nucleotide polymorphism RAD50 |
| title | The Association between Inefficient Repair of DNA Double-Strand Breaks and Common Polymorphisms of the HRR and NHEJ Repair Genes in Patients with Rheumatoid Arthritis |
| title_full | The Association between Inefficient Repair of DNA Double-Strand Breaks and Common Polymorphisms of the HRR and NHEJ Repair Genes in Patients with Rheumatoid Arthritis |
| title_fullStr | The Association between Inefficient Repair of DNA Double-Strand Breaks and Common Polymorphisms of the HRR and NHEJ Repair Genes in Patients with Rheumatoid Arthritis |
| title_full_unstemmed | The Association between Inefficient Repair of DNA Double-Strand Breaks and Common Polymorphisms of the HRR and NHEJ Repair Genes in Patients with Rheumatoid Arthritis |
| title_short | The Association between Inefficient Repair of DNA Double-Strand Breaks and Common Polymorphisms of the HRR and NHEJ Repair Genes in Patients with Rheumatoid Arthritis |
| title_sort | association between inefficient repair of dna double strand breaks and common polymorphisms of the hrr and nhej repair genes in patients with rheumatoid arthritis |
| topic | rheumatoid arthritis comet assay DNA double-strand break DNA repair single-nucleotide polymorphism RAD50 |
| url | https://www.mdpi.com/1422-0067/25/5/2619 |
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