Abstract: FR-PO292
Tubular Injury-Derived Exosome Interve Podocyte Cytoskeleton Rearrangement in Diabetic Mice
Session Information
- Diabetic Kidney Disease: Basic - 1
October 25, 2024 | Location: Exhibit Hall, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Diabetic Kidney Disease
- 701 Diabetic Kidney Disease: Basic
Authors
- Zhang, Rui, Department of Nephrology, Affiliated Hospital of Guizhou Medical University, Guiyang, China
- Yan, Rui, Department of Nephrology, Affiliated Hospital of Guizhou Medical University, Guiyang, China
Background
Renal tubular epithelial cells are the major source of exosomes in the kidney, carrying biological payload of proteins and nucleic acids to facilitate cellular communication. Here, we aimed to investigate the effect of renal tubule-derived exosomes on podocyte skeletal rearrangement in diabetic mice.
Methods
The mice renal tubular epithelial cells (mRETC), from which exosomes were purified and intravenously injected into wt mice, were cultured under normal glucose as control and high glucose combined with palmitic acid (HG+PA). MicroRNA (miRNAs) sequencing was performed to conduct whole-genome analysis of exosome miRNA by Illumina Hiseq2500. db/m mice and db/db mice were intravenously injected adeno-associated virus (AAV) vecor knockdown of miR-20a-5p and miRNA mimics overexpression (OE) of miR-20a-5p respectively. The expressions of nephrin, CD2AP, myosin X were detected by real-time PCR or westernblot. F-actin were detected by confocal microscopy. In vitro studies were performed in human podocyte cell tranfected with miRNA mimics overexpression of miR-20a-5p, podocyte cell were cultured with normal glucose, high glucose combined with palmitic acid.
Results
In WT mice with HG+PA exosome, renal histology showed podocyte foot process effacement by transmission electron microscope, nephrin, CD2AP, myosin X were significantly decreased compared with control mice. MiR-20a-5p was identified as one of the most regulated miRNAs in the exosomes from injure human kidney-2 (HK-2). The db/m mice treated with miR-20a-5p-OE exist more severely podocyte foot process effacement. In db/db mice treated with miR-20a-5p-AAV shRNA, renal histology showed foot process effacement decreased, expression level of nephrin, CD2AP, myosin X were significantly increased compared with db/db mice. In vitro the dual-luciferase reporter assay verified the binding of miR-20a-5p to MYH10 promoters, mutant MYH10 gene had downregulated nephrin, CD2AP expression and exhibited F-actin as polymerized state.
Conclusion
In diabetic mice, the expression of miR-20a-5p in exosomes from renal tubular epithelial cells is elevated. This miR-20a-5p mediates cytoskeletal rearrangement in podocytes by inhibiting myosin X, leading to foot process effacement. Consequently, targeting exosomes could represent a novel therapeutic approach for treating