National Clinical Research Center for Metabolic Diseases, Key Laboratory of Diabetes Immunology (Central South University), Ministry of Education, Department of Metabolism and Endocrinology, The Second Xiangya Hospital of Central South University, Changsha, Hunan, China.
Front Immunol. 2022 Nov 15;13:1030728. doi: 10.3389/fimmu.2022.1030728. eCollection 2022.
Type 1 diabetes mellitus (T1DM) is caused by immune cell-mediated β-cell dysfunction. In recent decades, N6-methyladenosine (m6A) has attracted widespread attention in the scientific research field because it plays vital roles in the pathogenesis of immunity-related diseases, including autoimmune diseases. However, neither the m6A modification profile nor the potential role it plays in T1DM pathogenesis has been investigated to date.
An m6A mRNA epitranscriptomic microarray analysis was performed to analyze m6A regulator expression patterns and m6A methylation patterns in immune cells of T1DM patients (n=6) and healthy individuals (n=6). A bioinformatics analysis was subsequently performed to explore the potential biological functions and signaling pathways underlying T1DM pathogenesis. Furthermore, mRNA expression and m6A methylation levels were subsequently verified by qRT-PCR and methylated RNA immunoprecipitation-qPCR (MeRIP-qPCR), respectively, in the T1DM and healthy groups (n=6 per group).
Among the multiple m6A regulators, METTL3 and IGF2BP2 had significantly downregulated expression, and YTHDC1 and HNRNPA2B1 had significantly upregulated expression in the T1DM group relative to the healthy group. The microarray analysis revealed 4247 differentially methylated transcripts, including 932 hypermethylated and 3315 hypomethylated transcripts, and 4264 differentially expressed transcripts, including 1818 upregulated transcripts and 2446 downregulated transcripts in the T1DM group relative to the healthy group. An association analysis between methylation and gene expression demonstrated that the expression of 590 hypermethylated transcripts was upregulated, and that of 1890 hypomethylated transcripts was downregulated. Pearson correlation analysis showed significant correlations between the expression levels of differentially expressed m6A regulators and the methylation levels of differentially methylated transcripts and significant correlations between the expression levels of differentially expressed m6A regulators and that of differentially expressed transcripts. Moreover, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses demonstrated that differentially methylated transcripts were involved in pathways related to immunity, including some closely associated with T1DM.
Our study presents m6A regulator expression patterns and m6A methylation patterns of immune cells in T1DM, showing that the m6A mark and m6A regulators are promising targets for T1DM diagnosis and treatment.
1 型糖尿病（T1DM）是由免疫细胞介导的β细胞功能障碍引起的。近几十年来，N6-甲基腺苷（m6A）因其在包括自身免疫性疾病在内的免疫相关疾病的发病机制中发挥着重要作用而引起了科学界的广泛关注。然而，目前尚未研究 m6A 修饰谱及其在 T1DM 发病机制中的潜在作用。
我们进行了 m6A mRNA 表转录组微阵列分析，以分析 T1DM 患者（n=6）和健康个体（n=6）免疫细胞中的 m6A 调节因子表达模式和 m6A 甲基化模式。随后进行了生物信息学分析，以探讨 T1DM 发病机制的潜在生物学功能和信号通路。此外，我们还通过 qRT-PCR 和甲基化 RNA 免疫沉淀-qPCR（MeRIP-qPCR）分别在 T1DM 和健康组（每组 n=6）中验证了 mRNA 表达和 m6A 甲基化水平。
在多种 m6A 调节剂中，METTL3 和 IGF2BP2 的表达明显下调，而 YTHDC1 和 HNRNPA2B1 的表达明显上调。微阵列分析显示，与健康组相比，T1DM 组有 4247 个差异甲基化转录本，其中 932 个呈高甲基化，3315 个呈低甲基化，有 4264 个差异表达转录本，其中 1818 个呈高表达，2446 个呈低表达。在 T1DM 组中，与健康组相比，有 590 个高甲基化转录本的表达上调，有 1890 个低甲基化转录本的表达下调。Pearson 相关性分析显示，差异表达 m6A 调节剂的表达水平与差异甲基化转录本的甲基化水平之间存在显著相关性，差异表达 m6A 调节剂的表达水平与差异表达转录本之间也存在显著相关性。此外，GO 和 KEGG 通路分析表明，差异甲基化转录本参与了与免疫相关的途径，其中一些途径与 T1DM 密切相关。
我们的研究展示了 T1DM 免疫细胞中的 m6A 调节因子表达模式和 m6A 甲基化模式，表明 m6A 标记和 m6A 调节剂是 T1DM 诊断和治疗的有前途的靶点。
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