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Kidney Week

Abstract: FR-PO735

Extracellular Vesicle and Modulation of miR-93 in Kidney Diseases

Session Information

Category: Glomerular Diseases

  • 1401 Glomerular Diseases: Mechanisms, including 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
  • Villani, Valentina, Children's Hospital Los Angeles, Los Angeles, California, United States
  • Hou, Xiaogang, Children's Hospital Los Angeles, Los Angeles, California, United States
  • Neviani, Paolo, Children's Hospital Los Angeles, Los Angeles, California, United States
  • Clair, Geremy, Pacific Northwest National Laboratory, Richland, Washington, United States
  • Alcala, Velia, Children's Hospital Los Angeles, Los Angeles, California, United States
  • Aguiari, Paola, Children's Hospital Los Angeles, Los Angeles, California, United States
  • Orlando, Giuseppe, Atrium Health Wake Forest Baptist Medical Center, Winston-Salem, North Carolina, United States
  • De Filippo, Roger E., Children's Hospital Los Angeles, Los Angeles, California, United States
  • Da Sacco, Stefano, Children's Hospital Los Angeles, Los Angeles, California, United States
  • Bussolati, Benedetta, University of Torino, Torino, Italy
  • Sedrakyan, Sargis, Children's Hospital Los Angeles, Los Angeles, California, United States
Background

Modulation of miRNA expression in glomerular cells is associated with renal disease. MiR-93, a potent regulator of glomerular damage, is downregulated in glomeruli of mice with Alport syndrome (AS, a renal disease characterized by a mutation in col4a5 gene) and in glomeruli of AS patients. Here, we investigated the role of extracellular vesicles (EVs) from human amniotic fluid stem cells (hAFSC) in disease-modifying activity in vitro and in vivo by regulation of miR-93 and its targets.

Methods

AS and WT glomeruli were characterized by bulk RNA-seq. hEVs, isolated by ultracentrifugation, were characterized by Exoview, TEM, super resolution microscopy, miRNA-seq, and proteomics. Expression of miR-93 was silenced in hEVs using a specific miR-93 antigomiR (KD-hEVs). Transfer and modulation of miR-93 (and its targets) in human glomerular cells was evaluated in vitro. hEV disease-modifying activity was evaluated by bulk RNA-seq in vitro, in AS mice by biodistribution, renal function, survival, and by spatial transcriptomics (ST) using the Visium 10X Genomics. Data were confirmed in AS patient biopsy using CosMX SMI (Nanostring).

Results

Glomerular endothelial cells are responsible for downregulation of miR-93 along disease progression. RNA-seq revealed alteration of miR-93 targets in AS glomeruli. Cell damage decreased expression of miR-93 and its targets, which were prevented by normal hEVs and not by KD-hEVs. hEV proteomics and miR-seq identified specific cargo and pathways central to disease progression. When injected in AS mice, hEVs localized in the kidney, ameliorated proteinuria, and prolonged lifespan compared to KD-hEVs. ST data showed that hEV injection upregulated important pathways responsible for glomerular regeneration, slowing down AS progression. Analysis of CosMX SMI also showed alteration of miR-93 targets in human AS patients.

Conclusion

hEVs modulated pathways that are central to glomerular homeostasis and improved kidney function by miR-93 transfer. These data suggest hEVs and modulation of miR-93 signaling might be important in translational settings.

Funding

  • NIDDK Support