mtDNA damage, mtDNA mutations and Calpain 10 (CAPN10) gene expression status in type 2 diabetes patients
Abstract
In this study, we aimed to analyze mitochondrial DNA (mtDNA) mutations, mtDNA damage and expression level of the Calpain 10 (CAPN10) gene in patients with Type-2 diabetes (T2D). Whole blood was drawn from 45 healthy participants who did not have diabetes mellitus and additional comorbid disease and 52 people with T2D for this investigation. DNA and RNA isolations of all samples were performed. For the analysis of mutations in the ATP6, ND1, CYB and D310 mtDNA genes, samples were first amplified by PCR. It was then confirmed by Sanger DNA sequencing. mtDNA -79 and mtDNA-230 fragments were amplified by RT-PCR to detect mtDNA damage. RT-PCR was also used for the mRNA expression of the CAPN10 gene. In patients with type-2 diabetes, m.8860 AG (ATP6) (52/52), m.15326 AG (CYB) (45/52), m.3384 AT (ND1) (2/52) and m.489 TC (D310) (11/52) were the most common mtDNA mutations. In addition, 7C (17/52), 8C (8/52) and 9C (1/52) mononucleotide repeats were detected in the D310 control region. Some of the discovered mutations were linked to damaging, illness causing, or benign characteristics, according to in silico analyses. An increase in the level of mtDNA-79, mtDNA-230 fragments and mtDNA integrity was determined in patients with T2Dcompared to healthy individuals (p=0.0090, p=0.9555, p=0.1213 respectively). The level of mtDNA-79 and mtDNA-230 fragments in patients showed a weak but significant positive correlation (p=0.0321, r=0.2977). In T2D patients compared to healthy people, the CAPN10 gene's mRNA expression was significantly higher (p=0.0360). Additionally, ROC analysis revealed that CAPN10 and mtDNA-79 had good diagnostic value in the patient group (AUC:0.603, p=0.0116, 95% CI:0.485-0.713, sensitivity: 53.8% for CAPN10 and AUC:0.653, p=0.0061, 95% CI:0.550-0.747, sensitivity: 51.9% for mtDNA-79). The findings showed that the frequency of mtDNA mutations and the expression level of the CAPN10 gene are high in patients with type-2 diabetes, and mtDNA damage is also increased. We believe that studying these mechanisms in the larger T2D patient population may be valuable in following the course of the disease.
Keywords
Type-2 diabetes, CAPN10, mtDNA, mtDNA-79, mtDNA-230, Type-2 diabetes, CAPN10, mtDNA, mtDNA-79, mtDNA-230
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