# Extracellular vesicle miR-93-5p cargo regulates glomerular endothelial cell damage in Alport syndrome

## TL;DR

This study shows that miR-93-5p in extracellular vesicles from stem cells can help repair kidney damage in Alport syndrome by restoring endothelial cell function.

## Contribution

The study identifies miR-93-5p as a key cargo in extracellular vesicles that mediates their therapeutic effect in Alport syndrome.

## Key findings

- miR-93-5p is downregulated in glomerular endothelial cells during Alport syndrome progression.
- hAFSC-EVs restore kidney function by transferring miR-93-5p and regulating VEGFR1 and VEGFR2 signaling.
- Spatial transcriptomics confirmed that hAFSC-EVs reverse disease pathways in the glomerulus via miR-93-5p.

## Abstract

Modulation of miRNA expression in glomerular cells is associated with renal disease. Here, we investigated the role of miR-93-5p in mitigating glomerular damage in Alport syndrome and whether the disease-modifying activity of extracellular vesicles from human amniotic fluid stem cells (hAFSC-EVs) is mediated by their miR-93-5p cargo. We identified downregulation of miR-93-5p specifically in glomerular endothelial cells in Alport syndrome along disease progression. Silencing of miR-93-5p in hAFSC-EVs changed the transcriptomic and proteomic profile, regulating EV disease-modifying activity. Compared with naive hAFSC-EVs, silenced hAFSC-EVs did not rescue glomerular endothelial function in vitro and did not restore kidney function in vivo. We established that hAFSC-EVs regulate VEGFR1 and VEGFR2 signaling by miR-93-5p cargo transfer, highlighting that miR-93-5p can restore glomerular endothelial cell biology. Spatial transcriptomics analysis of hAFSC-EV–injected kidneys showed that these EVs can reverse pathways altered during disease progression by stimulating proregenerative processes, specifically in the glomerulus, by regulating miR-93-5p targets. Alteration of glomerular endothelial cell transcriptomics and miR-93-5p targets was also confirmed in biopsies of patients with Alport syndrome using spatial molecular imaging. We demonstrated the critical role of miR-93-5p in glomerular endothelial cells and the capability of hAFSC-EVs to regulate miR-93-5p and its targets in Alport syndrome.

miR-93-5p-specific human AFSC-EVs cargo restores glomerular endothelial function in Alport syndrome by modulating VEGFR signaling and by promoting glomerular repair.