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Abstract：
To explore the role of lncRNA m6A methylation modification in aqueous humour (AH) of patients with pseudoexfoliation glaucoma (PXG). Patients with open-angle PXG under surgery from June 2021 to December 2021 were selected. Age- and gender-matched patients with age-related cataract (ARC) were chosen as control. Patients underwent detailed ophthalmic examinations. 0.05-0.1 ml AH were extracted during surgery for MeRIP-Seq and RNA-Seq. Joint analysis was used to screen lncRNAs with differential m6A methylation modification and expression. Online software tools were used to draw lncRNA-miRNA-mRNA network (ceRNA). Expression of lncRNAs and mRNAs was confirmed using quantitative real-time PCR. A total of 4151 lncRNAs and 4386 associated m6A methylation modified peaks were identified in the PXG group. Similarly, 2490 lncRNAs and 2595 associated m6A methylation modified peaks were detected in the control. Compared to the ARC group, the PXG group had 234 hypermethylated and 402 hypomethylated m6A peaks, with statistically significant differences (| Fold Change (FC) |≥2, p < 0.05). Bioinformatic analysis revealed that these differentially methylated lncRNA enriched in extracellular matrix formation, tight adhesion, TGF- β signalling pathway, AMPK signalling pathway, and MAPK signalling pathway. Joint analysis identified 10 lncRNAs with differential m6A methylation and expression simultaneously. Among them, the expression of ENST000000485383 and ROCK1 were confirmed downregulated in the PXG group by RT-qPCR. m6A methylation modification may affect the expression of lncRNA and participate in the pathogenesis of PXG through the ceRNA network. ENST000000485383-hsa miR592-ROCK1 May be a potential target pathway for further investigation in PXG m6A methylation.
摘要：
探讨lncRNAm6A甲基化修饰在假性剥脱性青光眼(PXG)患者房水(AH)中的作用。选择2021年6月至2021年12月接受手术的开角PXG患者。选择年龄和性别匹配的年龄相关性白内障（ARC）患者作为对照。患者接受了详细的眼科检查。在手术期间提取0.05-0.1mlAH用于MeRIP-Seq和RNA-Seq。联合分析用于筛选具有差异m6A甲基化修饰和表达的lncRNAs。使用在线软件工具绘制lncRNA-miRNA-mRNA网络(ceRNA)。使用定量实时PCR确认lncRNA和mRNA的表达。在PXG组中鉴定了总共4151个lncRNA和4386个相关的m6A甲基化修饰峰。同样，在对照中检测到2490个lncRNAs和2595个相关的m6A甲基化修饰峰。与ARC组相比，PXG组有234个高甲基化和402个低甲基化的m6A峰，差异有统计学意义(|倍数变化(FC)|≥2，p<0.05)。生物信息学分析显示，这些差异甲基化的lncRNA富集在细胞外基质形成中，紧密粘合,TGF-β信号通路,AMPK信号通路,和MAPK信号通路。联合分析鉴定了10个同时具有差异m6A甲基化和表达的lncRNAs。其中,RT-qPCR证实ENST000000485383和ROCK1的表达在PXG组中下调。m6A甲基化修饰可能影响lncRNA的表达，并通过ceRNA网络参与PXG的发病机制。ENST000000485383-hsamiR592-ROCK1可能是进一步研究PXGm6A甲基化的潜在靶途径。
