Identification and Validation of miR-206-3p Targeting WT-1 Promotes Membranous Nephropathy Through a Comprehensive...

Membranous nephropathy (MN), a major cause of end-stage renal disease, has limited therapeutics due to unclear targets. MicroRNAs (miRs) play critical roles in various diseases, but their role in MN remains unknown. This study utilizes a series of public databases, bioinformatics tools, and validation methods to identify key miRs and molecular mechanisms involved in MN. The GEO, miRbase, and TargetScan databases, along with the R package, Bioinformatics and ImageGP platform, were utilized to identify biomarkers in MN. 13 upregulated differentially expressed miRs (DEmiRs) were found across datasets. Weighted gene co-expression network analysis (WGCNA) showed that 58 miRs were strongly positively correlated with MN. Among these, 11 overlapping miRs were analyzed using the least absolute shrinkage and selection operator (LASSO) method to identify 5 candidate hub miRs. A series of approaches, including target gene prediction, disease-related analysis, and machine learning, brought miR-206 and WT-1 into focus as biomarkers for MN. Then, the passive Heymann nephropathy rat model and the adriamycin-injured podocytes were developed to validate these analyses and explore potential mechanisms. The gene or protein expression of upregulated miR-206-3p and downregulated WT-1 was verified in apoptotic podocytes both in vivo and in vitro. Moreover, the C-MIP/Nephrin/PI3K pathway appeared to be the potential mechanism through which miR-206-3p targets WT-1 to mediate podocyte apoptosis in MN. Ultimately, inhibition of miR-206-3p restored WT-1 expression and attenuated podocyte apoptosis. In summary, this study sheds light on the role of miR-206-3p targeting WT-1 in MN and confirms its involvement in podocyte apoptosis via the C-MIP/Nephrin/PI3K pathway.