Abstract: FR-PO691
Extracellular Vesicle and Modulation of miR-93 in Kidney Disease
Session Information
- Glomerular Diseases: Podocyte Biology - I
November 04, 2022 | Location: Exhibit Hall, Orange County Convention Center‚ West Building
Abstract Time: 10:00 AM - 12:00 PM
Category: Glomerular Diseases
- 1304 Glomerular Diseases: Podocyte Biology
Authors
- Perin, Laura, Children's Hospital Los Angeles, Los Angeles, California, United States
- Dedhia, Charmi, Children's Hospital Los Angeles, Los Angeles, California, United States
- Neviani, Paolo, Children's Hospital Los Angeles, Los Angeles, California, United States
- Aguiari, Paola, Children's Hospital Los Angeles, Los Angeles, California, United States
- Villani, Valentina, Children's Hospital Los Angeles, Los Angeles, California, United States
- Petrosyan, Astgik, Children's Hospital Los Angeles, Los Angeles, California, United States
- Hou, Xiaogang, Children's Hospital Los Angeles, Los Angeles, California, United States
- Bussolati, Benedetta, Children's Hospital Los Angeles, Los Angeles, California, United States
- Sedrakyan, Sargis, Children's Hospital Los Angeles, Los Angeles, California, United States
Background
Modulation of miRNA expression in glomerular cells is associated with renal diseases. Our data indicate that miR-93 is down-regulated in glomeruli of mice with Alport syndrome (AS, our model of the renal disease characterized by a mutation in coll4a5 gene) and in glomeruli of AS patients. Here, we investigated the role of hEVs derived from human amniotic fluid stem cells (hAFSC) in disease-modifying activity in vitro and in vivo by regulation of miR-93.
Methods
Isolated hEVs were characterized by cytometry, Exoview, immune-modulatory activity, RNA-seq, and proteomics. KO hEVs for miR-93 were generated by the use of an antagomir. Transfer and modulation of miR-93 (and its targets) in damaged human glomerular cells were evaluated in vitro. hEV disease-modifying activity was evaluated in AS mice by biodistribution, renal function, survival, and spatial transcriptomics (ST, Visium 10X Genomics Platform).
Results
By in situ analysis, we determined that miR-93 expression is decreased in mouse and human AS glomeruli. Cell specific damage decreases expression of miR-93 and its target expression (as VEGF) in human glomerular endothelial cells and podocytes. Expression of miR-93 was restored to normal levels only by normal hEVs and not KO hEVs. Proteomics and miR-seq data identified the specific hEV cargo fingerprint and that between the hEV and KO EV, a total of 59 proteins and 75 miRs were DE. GO and KEGG analysis of these cargo differences between hEVs and KO hEVs identified pathways central to disease progression, thus suggesting the central role of miR-93a in glomerular damage. When injected in AS mice, hEVs localized in the kidney, corrected proteinuria and prolong the lifespan. No side effects were noted. We defined spatial transcriptomics maps of kidneys from WT mice, AS mice, AS mice injected with hEVs and sacrificed after 5d and after 2m. Analysis of ST data on glomeruli showed that both early and late injections restored to normal important pathways responsible for AS progression (collagen formation, extracellular matrix alteration, fatty acid alteration) including miR-93 target pathways
Conclusion
hEVs regulated pathways that are central to glomerular homeostasis by modulation of miR-93. This suggests the possibility of using hEVs as a new therapeutic option for treating AS.
Funding
- NIDDK Support