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Article Abstract
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.
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| http://dx.doi.org/10.1016/j.gene.2025.149648 | DOI Listing |
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