糖尿病肾病中miRNA 5-甲基胞嘧啶修饰谱特征及潜在免疫相关靶基因调控
Signatures of miRNA 5-methylcytosine modification profile and potential immune-related target gene regulation in diabetic kidney disease.
作者信息
Yang Zhiqian, Yang Yan, Guo Hui, Gong Wenyu, Qiu Jing, Yan Qiang, Chen Huaizhou, Li Haitao, Zeng Zhipeng, Liu Fanna, Dai Yong, Tang Donge
机构信息
Guangdong Provincial Autoimmune Disease Precision Medicine Engineering Research Center, Shenzhen Autoimmune Disease Engineering Research Center, Shenzhen Geriatrics Clinical Research Center, Shenzhen People's Hospital, Second Clinical Medical College of Jinan University, Shenzhen 518020, China; Department of Nephrology, The First Affiliated Hospital of Jinan University, Guangzhou 510632, China.
Department of General Practice, Puning People's Hospital, Puning 515300, China.
出版信息
Gene. 2025 Sep 10;964:149648. doi: 10.1016/j.gene.2025.149648. Epub 2025 Jun 25.
BACKGROUND
Diabetic kidney disease (DKD) is a major microvascular complication of diabetes, with microRNAs (miRNAs) playing a role in its pathogenesis through methylation modifications. Preliminary experiments indicated an association between 5-methylcytosine (m5C) modification and miRNA dysregulation in peripheral blood mononuclear cells (PBMCs) of DKD patients.
METHODS
We employed Arraystar's small RNA modification chip to detect m5C modifications in miRNAs from PBMCs of DKD patients and predicted target genes. Transcriptomic data were analyzed for immune infiltration, and weighted gene co-expression network analysis (WGCNA) was performed to identify key target genes. Validation was conducted using MeRIP-qPCR and quantitative real-time PCR.
RESULTS
In DKD patients, 17 miRNAs showed elevated m5C levels, while 102 miRNAs exhibited reduced levels, associated with autophagy, pancreatic hormone signaling, and immune regulation. Immune infiltration analysis revealed significant differences in the infiltration of monocytes and eosinophils, correlating with classical immune pathways. Through WGCNA analysis of key monocyte modules, combined with target gene prediction, we identified TGFBR2 as a key target gene for the differentially m5C-modified miRNAs, potentially influencing immune infiltration in DKD. Further validation showed decreased m5C modification of miR-654-5p and elevated TGFBR2 expression in DKD, with a significant negative correlation. This relationship was further supported by qPCR analysis showing key methyltransferases (DNMT3B and NSUN2) expression inversely correlated with TGFBR2, suggesting m5C-mediated regulation.
CONCLUSIONS
Differential m5C modification of miRNAs may influence immune cell infiltration and contribute to the progression of DKD by modulating the expression of their target genes. TGFBR2 may serve as a target gene of miR-654-5p, regulated by its m5C modification status.
背景
糖尿病肾病（DKD）是糖尿病的一种主要微血管并发症，微小RNA（miRNA）通过甲基化修饰在其发病机制中发挥作用。初步实验表明，DKD患者外周血单个核细胞（PBMC）中5-甲基胞嘧啶（m5C）修饰与miRNA失调之间存在关联。
方法
我们使用Arraystar的小RNA修饰芯片检测DKD患者PBMC中miRNA的m5C修饰，并预测靶基因。对转录组数据进行免疫浸润分析，并进行加权基因共表达网络分析（WGCNA）以鉴定关键靶基因。使用MeRIP-qPCR和定量实时PCR进行验证。
结果
在DKD患者中，17种miRNA的m5C水平升高，而102种miRNA的水平降低，这与自噬、胰腺激素信号传导和免疫调节相关。免疫浸润分析显示单核细胞和嗜酸性粒细胞的浸润存在显著差异，与经典免疫途径相关。通过对关键单核细胞模块的WGCNA分析，结合靶基因预测，我们确定转化生长因子β受体2（TGFBR2）是m5C修饰差异的miRNA的关键靶基因，可能影响DKD中的免疫浸润。进一步验证显示，DKD中miR-654-5p的m5C修饰降低，TGFBR2表达升高，两者呈显著负相关。qPCR分析进一步支持了这种关系，显示关键甲基转移酶（DNMT3B和NSUN2）的表达与TGFBR2呈负相关，提示m5C介导的调控。
结论
miRNA的差异m5C修饰可能影响免疫细胞浸润，并通过调节其靶基因的表达促进DKD的进展。TGFBR2可能作为miR-654-5p的靶基因，受其m5C修饰状态调控。
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