Urinary Extracellular Vesicles and Their miRNA Cargo in Patients with Fabry Nephropathy

Abstract

Current biomarkers of Fabry nephropathy lack sensitivity in detecting early kidney damage and do not predict progression of nephropathy. Urinary extracellular vesicles (uEVs) and their molecular cargo could reflect early changes in renal impairment as they are secreted by the cells lining the urinary tract. We aimed to conduct a proof-of-concept study to investigate whether analysis of uEV characteristics and expression of uEV-derived microRNAs (miRNAs) could be applicable in studies to predict the development and progression of nephropathy in Fabry disease. A total of 20 Fabry patients were divided into two groups, depending on the presence of nephropathy. Chronological urine samples collected during 10-year follow-up were used for uEVs isolation with size exclusion chromatography. Nanoparticle tracking analysis was used to determine concentration and size of uEVs. We evaluated the expression of five uEV-derived miRNAs by qPCR (miR-23a-3p, miR-29a-3p, miR-30b-5p, miR-34a-5p, miR-200a-3p). There was no difference in the concentration and size of uEVs between patients with and without nephropathy at last follow-up or longitudinally. However, we found increased expression of miR-29a-3p and miR-200a-3p in uEVs isolated from chronological samples of patients with Fabry nephropathy. This may indicate an attempt by the organism to prevent the progression of renal damage leading to end-stage renal disease as previously reported in type 1 diabetes. In addition, we found an increased expression of miR-30b-5p in the 10-year period in uEVs of patients without renal dysfunction. miR-30b-5 was reported to have a protective role in podocyte injury and may possibly be important in Fabry nephropathy. These findings indicate that uEVs and their molecular cargo could be a promising target of studies focusing on elucidation of Fabry nephropathy. Nevertheless, total concentration and size of uEVs were neither indicative of the presence nor progression of Fabry nephropathy, while the role of the analyzed miRNAs in Fabry nephropathy progression was merely indicated and needs further in-depth studies.

Mitochondrial microRNAs Are Dysregulated in Patients with Fabry Disease

“…Alterations in over 100 circulating miRNAs were identified in blood samples from FD patients in treatment with ERT compared with untreated patients (Xiao et al, 2019). Increased expression of miRNA miR-29a-3p and miR-200a-3p were detected in urinary extracellular vesicles isolated from patients with FD nephropathy (Levstek et al, 2021) which associates with the progression of kidney damage (Maier et al, 2021); miR-let-7a and let-7d were significantly increased in FD after therapy and have been suggested as potential markers for enzyme activity and inflammation in FD patients (Maier et al, 2021). These data point at miRNA profiling as a powerful diagnostic tool of FD progression and drug testing, as well as indicators of the development of specific FD phenotypes.…”

Extracellular Vesicles in Kidney Diseases: Moving Forward

“…In this regard Lange and colleagues have shown that miRNA 21 levels are increased in uEVs in individuals with diabetes as well as a mouse model of glomerulonephritis [74]. Similarly, unique miRNA profiles in uEVs have been reported in individuals with membranous nephropathy [75], hypertension with albuminuria [76], diabetic kidney disease [77], [78], SLE [79], and Fabry nephropathy [80]. Recently, Luan et al reported changes in the profile of circular RNAs in uEVs from individuals with IgA nephropathy as compared to healthy subjects [81].…”

“…Similarly, unique miRNA profiles in uEVs have been reported in individuals with membranous nephropathy, 90 hypertension with albuminuria, 91 diabetic kidney disease, 92,93 SLE, 94 and Fabry nephropathy. 95 Recently, Luan et al 96 reported changes in the profile of circular RNAs in uEVs from individuals with IgA nephropathy compared with healthy subjects. Thus, both the protein and nucleic acid composition of EVs seems to be altered in glomerular disease.…”

A kaleidoscopic view of extracellular vesicles in lysosomal storage disorders

“…In general, the use of EVs could increase the sensitivity of biochemical assays by increasing the concentration of test material. If successful, this could improve both maternal and fetal patient management [113] .…”