The m6A RNA modification modulates gene expression and fibrosis-related pathways in hypertrophic scar [m6A-seq]

Purpose: To systematically analyze the overall m6A modification pattern in hyperplastic scars (HS). Methods: The m6A modification patterns in HS and normal skin (NS) tissues were described by m6A sequencing and RNA sequencing, and subsequently bioinformatics analysis was performed. The m6A-related RNA was immunoprecipitated and verified by real-time quantitative PCR. Results: The appearance of 14,791 new m6A peaks in the HS sample was accompanied by the disappearance of 7,835 peaks. The unique m6A-related genes in HS were thus associated with fibrosis-related pathways. We identified the differentially expressed mRNA transcripts in HS samples with hyper-methylated or hypo-methylated m6A peaks. Conclusion: This study is the first to map the m6A transcriptome of human HS, which may help clarify the possible mechanism of m6A-mediated gene expression regulation. Overall design: Si-NC (C, n=1) and Si-METTL3 (SI, n=1) samples

研究目的：系统解析增生性瘢痕（hyperplastic scars, HS）中整体的m6A（N6-methyladenosine）修饰谱型。
研究方法：通过m6A测序与RNA测序获取HS及正常皮肤（normal skin, NS）组织的m6A修饰谱型，随后开展生物信息学分析；同时利用免疫沉淀技术富集m6A相关RNA，并通过实时定量聚合酶链反应（real-time quantitative PCR, qPCR）进行验证。
研究结果：HS样本中新增14791个m6A峰，同时原有7835个m6A峰消失；HS中特有的m6A相关基因与纤维化相关通路显著关联；本研究鉴定出HS样本中携带高甲基化或低甲基化m6A峰的差异表达mRNA转录本。
研究结论：本研究首次绘制了人类增生性瘢痕的m6A转录组图谱，可为阐明m6A介导的基因表达调控潜在机制提供参考。
实验设计：设置阴性对照小干扰RNA组（Si-NC, C, n=1）与METTL3靶向小干扰RNA组（Si-METTL3, SI, n=1），每组各1例样本。