Abstract: SA-PO0263
NEDDylation in Proximal Tubular Cells Promotes Ischemia-Reperfusion-Induced AKI
Session Information
- AKI: Mechanisms - Cell Signaling
October 24, 2026 | Location: Exhibit Hall A, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Acute Kidney Injury
- 103 AKI: Mechanisms
Authors
- Wu, Yan, Augusta University, Augusta, Georgia, United States
- Dong, Zheng, Augusta University, Augusta, Georgia, United States
Background
Ischemia-reperfusion (I/R) injury is a leading cause of acute kidney injury (AKI), yet specific clinical interventions remain limited. NEDDylation, a ubiquitin-like post-translational modification catalyzed by NAE1 may regulate cellular stress and death. However, whether NEDDylation is involved in renal tubular epithelial cell I/R injury and AKI progression remains unclear.
Methods
In vitro, Rat proximal tubular epithelial cells (RPTCs) were subjected to sodium azide treatment(3h) followed by recovery(2h) in normal culture medium. In vivo, C57BL/6 mice underwent 30min bilateral renal ischemia with 48 h repercussion. NEDDylation was inhibited pharmacologically using MLN4924 and genetically via proximal tubule-specific NAE1 conditional knockout (NAE1 PT-KO). NEDDylation levels, tubular injury, and apoptosis were assessed by Western blot, IHC/IF, serum BUN/creatinine, HE staining, and morphological analysis.
Results
In the in vitro model, RPTCs exhibited elevated NEDDylation levels, evidenced by increased NAE1 protein expression and enhanced NEDD8-conjugated cullins, accompanied by elevated cleaved caspase-3 expression and increased apoptosis. MLN4924 treatment suppressed NEDD8-conjugated cullins, reduced apoptotic rates, and downregulated cleaved caspase-3 expression. In vivo, renal NAE1 and NEDD8-conjugated cullin levels were markedly elevated post-I/R, with IHC/IF demonstrating predominant NEDDylation signal enrichment in injured proximal tubules. MLN4924 administration decreased serum BUN and creatinine, ameliorated tubular pathological injury on HE staining, and reduced KIM-1 expression with concomitant suppression of NEDD8-conjugated cullins. Furthermore, proximal tubule-specific NAE1 conditional knockout mice were generated and validated. Following I/R injury, NAE1 PT-KO mice exhibited lower serum BUN and creatinine levels, attenuated tubular morphological injury, and reduced KIM-1 expression than wild-type mice.
Conclusion
I/R injury activates NEDDylation in proximal tubular epithelial cells to promote AKI progression. Pharmacological or genetic inhibition of NEDDylation effectively ameliorates kidney injury. NEDDylation may represent a potential therapeutic target in AKI.