Abstract: TH-PO988
miR-466o-3p Promotes Podocyte Injury Through Targeting WT1
Session Information
- Pathology and Lab Medicine: Basic
October 25, 2018 | Location: Exhibit Hall, San Diego Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Pathology and Lab Medicine
- 1501 Pathology and Lab Medicine: Basic
Authors
- Chen, Qiyan, Nanfang Hospital, Southern Medical University, Guangzhou, China
- Zhou, Lili, Nanfang Hospital, Southern Medical University, Guangzhou, China
- Liu, Youhua, University of Pittsburgh, Pittsburgh, Pennsylvania, United States
Background
Podocytes are highly specialized cells that maintain normal function of the glomerular filtration barrier. Dysregulation of Wnt/β-catenin signaling has been linked to podocyte injury; however, the underlying mechanism remains incompletely understood. In this study, we explored the roles of microRNA in mediating Wnt/β-catenin-triggered podocyte dysfunction and proteinuria.
Methods
Mouse podocytes were transiently transfected with expression vector encoding constitutively activated β-catenin (pDel-β-cat) or empty vector (pcDNA3). The differential expression of microRNA between these two groups was analyzed microarray assay and bioinformatics analysis. The role of miR-466o-3p in podocyte biology was investigated in vitro and in vivo. The BALB/c mice were randomly divided into 3 different groups: i) sham control; ii) ADR mice injected with empty vector pcDNA3; iii)ADR mice injected with miR-466o-3p plasmid. Urine, blood and kidneys were collected at 1 week after ADR injection.
Results
We identified multiple miRNAs whose expression was modulated by β-catenin activation in podocytes. Real-time RT-PCR validated that miRNA-466o-3p was upregulated in cultured podocytes after β-catenin activation and in kidney tissues after adriamycin (ADR) injection. In situ hybridization demonstrated that miR-466o-3p was up-regulated in glomerular podocytes. Bioinformatics analysis and luciferase reporter assay confirmed that miR-466o-3p directly targeted WT1 mRNA. Furthermore, overexpression of miR-466o-3p downregulated WT1 protein in vitro and promoted podocyte injury, although it did not affect WT1 mRNA level. Conversely, inhibition of miR-466o-3p alleviated β-catenin induced podocyte injury. In ADR model, Overexpression of miR-466o-3p inhibited WT1, aggravated podocytes injury and increased proteinuria in mice.
Conclusion
These studies demonstrate that miR-466o-3p plays a critical role in mediating Wnt/β-catenin-triggered podocyte injury. Our findings uncover a new pathogenic mechanism by which Wnt/β-catenin promotes podocyte injury and proteinuria.
Funding
- NIDDK Support