School of Public Health, Health Science Center, Key Laboratory of Environmental and Endemic Diseases of National Health Commission of the People's Republic of China, Xi'an Jiaotong University, Xi'an, People's Republic of China.
Shaanxi Provincial Institute for Endemic Disease Prevention and Control, Xi'an, People's Republic of China.
J Cell Mol Med. 2024 Oct;28(20):e70047. doi: 10.1111/jcmm.70047.
Kashin-Beck disease (KBD) is a chronic degenerative, disabling disease of the bones and joints and its exact aetiology and pathogenesis remain uncertain. This study is to investigate the role of m6A modification in the pathogenesis of KBD. Combined analysis of m6A MeRIP-Seq and RNA-Seq were used to analyse human peripheral blood samples from three KBD patients and three normal controls (NC). Bioinformatic methods were used to analyse m6A-modified differential genes and RT-qPCR was performed to validate the mRNA expression of several KBD-related genes. The results indicated that the total of 16,811 genes were modified by m6A in KBD group, of which 4882 genes were differential genes. A large number of differential genes were associated with regulation of transcription, signal transduction and protein binding. KEGG analysis showed that m6A-enriched genes participated the pathways of Vitamin B6 metabolism, endocytosis and Rap 1 signalling pathway. There was a positive association between m6A abundance and levels of gene expression, that there were 6 hypermethylated and upregulated genes (hyper-up), 23 hypomethylated and downregulated genes (hypo-down) in KBD group compared with NC. In addition, the mRNA expression of levels of MMP8, IL32 and GPX1 were verified and the protein-protein interaction networks of these key factors were constructed. Our study showed that m6A modifications may play a vital role in modulating gene expression, which represents a new clue to reveal the pathogenesis of KBD.
大骨节病（KBD）是一种慢性退行性、致残性骨和关节疾病，其确切病因和发病机制仍不清楚。本研究旨在探讨 m6A 修饰在 KBD 发病机制中的作用。采用 m6A MeRIP-Seq 和 RNA-Seq 联合分析方法，分析了 3 例 KBD 患者和 3 例正常对照（NC）的人外周血样本。利用生物信息学方法分析 m6A 修饰的差异基因，并采用 RT-qPCR 验证了几种与 KBD 相关的基因的 mRNA 表达。结果表明，KBD 组共有 16811 个基因被 m6A 修饰，其中 4882 个为差异基因。大量差异基因与转录调控、信号转导和蛋白质结合有关。KEGG 分析显示，m6A 富集基因参与了维生素 B6 代谢、内吞作用和 Rap1 信号通路等途径。m6A 丰度与基因表达水平呈正相关，与 NC 相比，KBD 组有 6 个高甲基化和上调基因（hyper-up）、23 个低甲基化和下调基因（hypo-down）。此外，还验证了 MMP8、IL32 和 GPX1 的 mRNA 表达水平，并构建了这些关键因子的蛋白质-蛋白质相互作用网络。我们的研究表明，m6A 修饰可能在调节基因表达中发挥重要作用，这为揭示 KBD 的发病机制提供了新的线索。
