Abstract：
N6-methyladenosine (m6A) modification is one of the most common types of methylation modifications in eukaryotic mRNA. However, its role in the pathogenesis of pseudoexfoliation glaucoma (PXG) has not yet been reported. To enhance understanding in this regard, we assessed the m6A methylome in the aqueous humor of patients with PXG. MeRIP-Seq and RNA-Seq analyses were performed to compare the m6A methylomes and gene expression profiles of the aqueous humor of patients with PXG with those of patients with age-related cataract (ARC). Colorimetric m6A quantification was performed to detect global m6A levels. Quantitative reverse transcription PCR confirmed the expression of m6A-related enzymes and mRNAs in both groups. Results showed significantly higher aqueous humor m6A levels in the PXG group than in the ARC group. Five m6A-related enzymes, including METTL3, YTHDC2, HNRNPA2B1, HNRNPC, and LRPPRC, were significantly up-regulated in PXG specimens. We also observed 9728 m6A-modified peaks related to 6126 gene transcripts in the PXG group, with more than 250 genes containing one m6A peak (hypomethylated or hypermethylated). The distribution of the m6A peaks was enriched in coding sequences and 3\'-untranslated regions for both groups. GGAC motif structures were also significantly enriched. Bioinformatics analysis further revealed that m6A plays a critical role in extracellular matrix formation and histone deacetylation. Additionally, MMP14, ADAMTSL1, FN1, and HDAC1 showed significant changes in m6A methylation and mRNA expression in the PXG group. Therefore, m6A methylation may regulate extracellular matrix composition in PXG and METTL3 may be a pivotal regulator of this process. In the future, it would be necessary to investigate MMP14, ADAMTSL1, FN1, and HDAC1, which are potential target genes.
摘要：
N6-甲基腺苷（m6A）修饰是真核mRNA中最常见的甲基化修饰类型之一。然而,其在假性剥脱性青光眼（PXG）发病机制中的作用尚未见报道。为了加强这方面的认识，我们评估了PXG患者房水中的m6A甲基化体。进行了MeRIP-Seq和RNA-Seq分析，以比较PXG患者与年龄相关性白内障（ARC）患者房水的m6A甲基化组和基因表达谱。进行比色m6A定量以检测全局m6A水平。定量逆转录PCR证实了两组中m6A相关酶和mRNA的表达。结果显示，PXG组的房水m6A水平明显高于ARC组。五种m6A相关酶，包括METTL3，YTHDC2，HNRNPA2B1，HNRNPC，和LRPPRC，在PXG标本中显著上调。我们还在PXG组中观察到与6126个基因转录本相关的9728个m6A修饰的峰，超过250个基因含有一个m6A峰（低甲基化或高甲基化）。m6A峰的分布在两个组的编码序列和3'-非翻译区中富集。GGAC基序结构也显著富集。生物信息学分析进一步表明，m6A在细胞外基质形成和组蛋白脱乙酰化中起关键作用。此外,MMP14、ADAMTSL1、FN1和HDAC1在PXG组中显示m6A甲基化和mRNA表达的显著转变。因此,m6A甲基化可以调节PXG中的细胞外基质组成，METTL3可能是该过程的关键调节因子。在未来,有必要研究潜在的靶基因MMP14,ADAMTSL1,FN1和HDAC1.
