DNA Methylation of Patatin-Like Phospholipase Domain-Containing Protein 6 Gene Contributes to the Risk of Intracranial Aneurysm in Males
Shengjun Zhou, Junjun Zhang, Chenhui Zhou, Fanyong Gong, Xueli Zhu, Xingqiang Pan, Jie Sun, Xiang Gao, Yi Huang
Abstract
Objective : This study is aimed to investigate the contribution of patatin-like phospholipase domain-containing protein 6 ( PNPLA6 ) DNA methylation to the risk of intracranial aneurysm (IA) in the Han Chinese population. Methods : A total of 96 age- and sex-matched participants were recruited to evaluate PNPLA6 methylation via bisulfite pyrosequencing. The PNPLA6 mRNA expression in the plasma was determined using real-time quantitative reverse transcription-polymerase chain reaction. Human primary artery smooth muscle cells (HPCASMC) were used for the in vitro function study. Results : PNPLA6 methylation was significantly higher in patients with IA than in healthy controls ( p < 0.01). Sex group analysis showed that this correlation appeared in the male group ( p < 0.01) but not in the female group ( p > 0.05). PNPLA6 methylation was significantly associated with age in all participants ( r = 0.306, p = 0.003) and in the control group ( r = 0.377, p = 0.008) but not in the IA group ( r = 0.127, p = 0.402). Furthermore, the PNPLA6 mRNA expression significantly decreased in patients with IA than that in the controls ( p = 0.016). PNPLA6 expression was significantly inversely correlated with elevated DNA methylation in participants (r = −0.825, p < 0.0001). In addition, PNPLA6 transcription was significantly enhanced following treatment with 5-aza-2’-deoxycytidine methylation inhibitor in HPCASMC.The receiver operating characteristic analyses of curves showed that the PNPLA6 mean methylation [area under the curve (AUC) = 0.74, p < 0.001] and mRNA expression (AUC = 0.86, p < 0.001) could have a diagnostic value for patients with IA. Conclusion : Although future functional experiments are required to test our hypothesis, our study demonstrated that PNPLA6 methylation and mRNA expression were significantly associated with the risk of IA; thus, they show potential for use in the early diagnosis of IA.