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Updated: May 28, 2025
Comparative Proteomic Analysis of Whole Kidney, Medulla, and Cortical Tubules in Diabetic Pathogenesis of Kidney Injury in Mice
Published on: May 2, 2025
Multiomics analysis of O-GlcNAcylation in podocytes of diabetic kidney disease
Yun Zou1,2,3, Mengyao Zhuo4, Wen Chen1
1Department of Endocrinology and Metabolism, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, People's Republic of China.
Aim:
To investigate the role of O-GlcNAc transferase (OGT)-mediated protein O-GlcNAcylation in podocyte injury during the progression of diabetic kidney disease (DKD).
Materials And Methods:
Proteomic and O-glycoproteomic analyses were conducted on high glucose (HG)-stimulated podocytes with OGT knockdown. Differentially expressed proteins and O-GlcNAcylated peptides/proteins were identified, and functional enrichment (GO, KEGG, COG/KOG) and motif analysis (motif-x) were performed using bioinformatics analysis. Co-immunoprecipitation (Co-IP) was used to validate O-GlcNAcylation of candidate proteins.
Results:
OGT knockdown in HG-treated podocytes resulted in 128 upregulated and 45 downregulated proteins. Glycoproteomics revealed 32 glycopeptides/21 glycoproteins upregulated and 37 glycopeptides/22 glycoproteins downregulated. The focus was on down-regulated glycosylated proteins without changes in their protein levels. These proteins are predominantly enriched in translation factor activity, RNA binding, and ECM-receptor interactions pathways. Among these proteins, Caprin1, Lrp1, and Sil1 were modified by O-GlcNAcylation.
Conclusion:
OGT-driven O-GlcNAcylation exacerbates podocyte injury in DKD by post-translationally modifying key regulators of translational machinery and ECM signalling. Precision targeting of O-GlcNAc dynamics represents a promising therapeutic strategy to attenuate DKD.
Insights
O-GlcNAc transferase (OGT)-mediated protein O-GlcNAcylation worsens podocyte injury in diabetic kidney disease (DKD). Targeting O-GlcNAc dynamics may offer a therapeutic strategy to reduce DKD progression.
Area of Science:
- Nephrology
- Molecular Biology
- Biochemistry
Background:
- Diabetic kidney disease (DKD) is a major complication of diabetes, characterized by progressive podocyte injury.
- Protein O-GlcNAcylation, regulated by O-GlcNAc transferase (OGT), plays a role in cellular signaling and disease pathogenesis.
Purpose of the Study:
- To investigate the role of OGT-mediated protein O-GlcNAcylation in podocyte injury during DKD progression.
- To identify specific proteins and pathways affected by O-GlcNAcylation in high glucose-induced podocyte injury.
Main Methods:
- Proteomic and O-glycoproteomic analyses of high glucose-stimulated podocytes with OGT knockdown.
- Bioinformatic analysis including functional enrichment and motif analysis.
- Co-immunoprecipitation to validate O-GlcNAcylation of candidate proteins.
Main Results:
- OGT knockdown altered protein and glycoprotein expression in high glucose-treated podocytes.
- Downregulated O-GlcNAcylated proteins were enriched in translation factor activity and ECM-receptor interaction pathways.
- Caprin1, Lrp1, and Sil1 were identified as O-GlcNAcylated proteins affected by OGT modulation.
Conclusions:
- OGT-driven O-GlcNAcylation exacerbates podocyte injury in DKD by modifying key regulators of translation and ECM signaling.
- Targeting O-GlcNAc dynamics presents a potential therapeutic avenue for attenuating DKD.
