#2922 The role of urinary extracellular-derived miRNAs dysregulation in IgA nephropathy

Abstract

Abstract Background and Aims IgA nephropathy (IgAN) is one of the most common forms of glomerulonephritis and causes hematuria and proteinuria by damaging the mesangial and podocyte cells. Urinary extracellular vesicles (uEVs) derived from damaged podocytes may reflect podocyte damage in various glomerular diseases and are considered an early and noninvasive diagnostic method. This study aimed to extract urinary extracellular vesicles (EVs) and investigate the expression dysregulation and diagnostic value of miR-29c, miR-146a, miR-148b, miR-155, miR-200b, and miR-429 in urine samples from patients with IgAN. Method Urine samples were collected from 20 patients with IgAN, 30 patients with membranous nephropathy (MGN), and 16 healthy individuals to extract Urinary EVs by ultracentrifugation. The characterization of EVs was evaluated by the specific protein markers of the EVs, flotillin-1, ALiX, and CD-9, along with histone H1 and podocyte-specific marker podocalyxin via western blotting. SEM and DLS were used to investigate the size and morphology of EVs. The expression levels of the miRNAs were evaluated using Real-time PCR. Results Numerous extracellular vesicles >100 nm in size were present in the urine sample which were considered as EVs based on references. There were higher levels of the specific markers flotillin-1, ALiX, and podococalyxin in the urine samples of IgAN patients than in the control samples (P < 0.001 and P < 0.001, respectively). There was a statistically significant difference among the expression level of miR-29c (P = 0.002 and P = 033) in the group of IgAN patients compared to MGN patients and the group of healthy people, based on Rock curve analysis this urinary marker could discriminate the IgAN patients from MGN group, the area under the curve AUC ≥ 0.73. Conclusion This study showed that the different levels of miRNAs studied in urinary EVs may indicate podocyte damage and have a direct relationship with the pathology of IgAN and MGN. Further studies are needed to confirm these results and determine the diagnostic value of these markers.