DNA Methylation of Patatin-Like Phospholipase Domain-Containing Protein 6 Gene Contributes to the Risk of Intracranial Aneurysm in Males

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.

Exploring the potential of VGLL3 methylation as a prognostic indicator for intracranial aneurysm with gender-specific considerations

“…YAP forward primer, 5′-ACAATGACGACCAATAGCTCAGAT-3′; YAP reverse primer, 5′-AACGGTTCTGCTGTGAGGG-3′. The sequence of the internal reference gene ACTB and the calculation method of mRNA expression have been described in our previous study [ 13 ].…”

“…Maimaiti et al used novel bioinformatics methods, including transcriptomics analysis, machine learning algorithms, genome-wide association studies (GWAS), Mendelian randomization (MR), and summary data-based Mendelian randomization (SMR), to demonstrate the association between DNA methylation-related genes DNMT3A, MBD2, DNMT1 , and DNMT3B and IA susceptibility [ 12 ]. Zhou et al found that DNA methylation levels are associated with the risk of IA in Chinese males [ 13 ].…”

Multi-omics in thoracic aortic aneurysm: the complex road to the simplification

“…Furthermore, increased methylation at phospholipase domain-containing protein 6 ( PNPLA6 ), leading to its consequent reduction in mRNA expression, was found to affect intracranial aneurysm development [ 73 ].…”

Nicotinic Acid-Mediated Modulation of Metastasis-Associated Protein 1 Methylation and Inflammation in Brain Arteriovenous Malformation

“…The primer sequences were as follows: MTA1 forward primer, 5 -TACCTGGAGCGGGAGGATTTCTT-3 ; and reverse primer, 5 -GCTTGTCTGTGAGTGGGTTGTGC-3 . ACTB was used as an internal reference [24], with the forward primer being 5 -AGCACAGAGCCTCGCCTT-3 and the reverse primer being 5 -CATCATCCATGGTGAGCTGG-3 . The 2 − Ct method was used to calculate mRNA expression.…”