mRNA expression and methylation rate of two antithrombotic genes, KLF2 and THBD, in patients with ischemic stroke and early onset ischemic stroke in comparison with healthy controls.

The most common type of stroke is ischemic stroke. Key regulators of thrombosis including KLF2, eNOS, TM and PAI-1 are considered as potential causes in ischemic stroke pathogenesis. This study examined gene expression and DNA methylation status of KLF2 and THBD genes in ischemic stroke patients. We selected 48 ischemic stroke patients and 48 healthy controls. Gene expression and methylation status of KLF2 and THBD genes were evaluated by real-time RT-PCR and MS-PCR, respectively. Clinical characteristics were also evaluated based on the methylation status of the patients. Statistical analysis was done using SPSS-26 software. mRNA expression of KLF2 (P = 0.0157) and THBD (P = 0.0070) was significantly reduced in ischemic stroke compared to the control. The hypermethylation of KLF2 and THBD genes significantly increased in ischemic stroke group (P = 0.026 and P = 0.021, respectively). Moreover, significantly increased methylation of KLF2 and THBD genes were seen in early onset ischemic stroke group than control group (P < 0.05). Hypermethylation in KLF2 and THBD was significantly associated with the occurrence of global aphasia. Also, hypermethylation in KLF2 was associated with dysarthria. The binary logistic regression analysis showed that KLF2 methylation, THBD methylation, obesity and asthma significantly and independently associated with the occurrence of ischemic stroke. However, diabetes and smoking revealed borderline association. Our study identified hypermethylation in KLF2 and THBD genes as a novel risk marker for ischemic stroke development. So, appropriate management of hypermethylation in KLF2 and THBD genes may be considered as therapeutic strategies for ischemic stroke patients.