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Abstract: PO1987

Glomerular Heterogeneity and Modulation of miR-93: The Role of Extracellular Vesicles

Session Information

  • Podocyte Biology
    October 22, 2020 | Location: On-Demand
    Abstract Time: 10:00 AM - 12:00 PM

Category: Glomerular Diseases

  • 1204 Podocyte Biology

Authors

  • Perin, Laura, Children's Hospital of Los Angeles, Los Angeles, California, United States
  • Dedhia, Charmi, Children's Hospital of Los Angeles, Los Angeles, California, United States
  • Aguiari, Paola, Children's Hospital of Los Angeles, Los Angeles, California, United States
  • De Filippo, Roger E., Children's Hospital of Los Angeles, Los Angeles, California, United States
  • Sedrakyan, Sargis, Children's Hospital of Los Angeles, Los Angeles, California, United States
Background

Modulation of miRNA in podocytes and glomerular endothelial cells (GEC) has been associated with development of renal diseases. miR-93 is a potent regulator of pathways responsible for glomerular damage, like VEGF, TGFb and Msk2. We have evidence that miR-93 is altered in the glomeruli of mice with X-linked Alport syndrome (AS), carrying the Col4a5 mutation, and in glomeruli of AS patients. Here, we investigated the role of miR-93 in mesangial cells, podocytes and GEC from WT and AS mice. We also used extracellular vesicles (hEVs) derived from human amniotic fluid stem cells (hAFSC) to assess their disease modifying activity in vitro and in vivo by regulation of miR-93.

Methods

miR-93 expression was evaluated by qRT-PCR in mesangial cells, podocytes and GEC sorted from glomeruli of male and female WT (C57BL6/J), and homozygous and heterozygous AS (Col4a5-/-) mice at different stages of disease (2m, 3.5m and 5.5m) and in biopsies of AS patients. Modulation of miR-93 by hEVs was evaluated in vitro and EV therapeutic effect was evaluated in vivo by RNA-seq and survival.

Results

miR-93 expression is different between male and female mice along disease progression. In AS males miR-93 level was significantly lower in GEC, but not in podocytes or mesangial cells vs WT cells. miR-93 expression was downregulated also in AS patients. Expression of WT1 in puromycin aminonucleoside damaged podocytes, and expression of fibronectin and VEGF in damaged GEC was restored by miR-93 hEV cargo transfer. In vivo, hEVs showed therapeutic effect by ameliorating the level of proteinuria and increasing life span. Transcriptomic analysis showed that WT male and female present differences in respiratory and metabolic pathways, extracellular matrix and cell adhesion molecules. AS males injected with hEVs showed improved gene modulations in metabolic function and in the development of vasculature, angiogenesis and fibrosis pathways, important miR-93 targets.

Conclusion

Gender specific variation in miR-93 expression in glomerular cells might indicate important differences in response to injury in progressive disease. hEVs demonstrate great potential to restore lost miR-93 expression and its targets, thus presenting a targeted approach for treatment of CKD.

Funding

  • NIDDK Support