Integrated Bioinformatics and Experimental Validation Reveal Shared Molecular Targets for Diagnosis and Intervention in Type 2 Diabetes Mellitus and Diabetic Nephropathy.
Abstract
INTRODUCTION Type 2 diabetes mellitus (T2DM) is a major risk factor for diabetic nephropathy (DN), yet the molecular mechanisms connecting these conditions remain unclear. Identifying shared hub genes and regulatory networks may provide insight into common pathogenic pathways.
Methods
Four GEO microarray datasets associated with T2DM (GSE23343, GSE29226) and DN (GSE30528, GSE142153) were analyzed using limma in R to identify differentially expressed genes (DEGs). Overlapping DEGs were assessed using Venn analysis and integrated into a STRING-based protein-protein interaction network. Hub genes were identified in Cytoscape. Their expression under high-glucose conditions was validated in HK-2 and NRK-52E cells using RT-qPCR and Western blotting. Predicted miRNAs were obtained from TargetScan and evaluated experimentally. Functional assays assessed the effects of hub gene overexpression.
Results
Thirty-six common DEGs were identified, with APP, RHEB, FRYL, and SOS1 exhibiting highest network connectivity. All four genes were consistently downregulated in patient datasets and high-glucose cell models. ROC analyses indicated moderate discriminatory capacity within datasets. Four candidate miRNAs (miR-26b-5p, miR-18a-5p, miR-199a-5p, miR-148a-3p) were elevated under highglucose conditions. Functional enrichment linked hub genes to mTOR, PI3K-Akt, and cytoskeletal pathways. Overexpression of hub genes reduced proliferation, clonogenicity, and migration in vitro.
Discussion
The convergence of transcriptomic and experimental findings suggests that reduced expression of these hub genes may contribute to glucose-induced cellular dysfunction. However, miRNA-gene relationships remain correlative, and validation in additional renal cell types and independent patient cohorts is needed.
Conclusion
APP, RHEB, FRYL, and SOS1 represent shared molecular signatures of T2DM and DN and may offer potential targets for future mechanistic and therapeutic studies.