Introduction: Urinary exosomes are associated with kidney stones but how they work is unclear. In this study, we aim to identify dysregulated miRNAs in urinary exosomes from kidney stone patients and to explore the potential role of exosomal miRNAs in nephrolithiasis.
Methods: First morning voids were collected from participants. Urinary exosomes were isolated via sequential differential centrifugation and ultracentrifugation. Next-generation sequencing was conducted to analyze the transcriptome of urine exosomes from three kidney stone patients and three age-/sex-matched healthy controls. Bioinformatics analysis was performed to explore how dysregulated miRNAs were associated with stone formation. The miRNA sequencing results of exosomes derived from epithelial cells(GSE110509) were compared with the dysregulated miRNAs of our study, and finally we found three common miRNAs. The three miRNAs were verified by qPCR in other five kidney stone patients and five healthy controls.
Results: With the screening criteria of absolute log2 fold change(log2FC)>1 and P<0.01, 54 dysregulated miRNAs were found between control and stone groups, among which 14 miRNAs were down-regulated and 40 miRNAs were up-regulated in stone groups compared to control groups. Gene ontology (GO) analysis indicated that dysregulated miRNAs were enriched in oxidative stress(Biological Process),focal adhesion(Cellular Component),and cell adhesion molecule binding(Molecular Function).KEGG pathways analysis demonstrated that dysregulated miRNAs were common enriched in MAPK signaling pathway, focal adhesion and AGE-RAGE signaling pathway. The expression of miR-223-3p,miR-127-3p and miR-214-5p were significantly different in either the exosomes derived from cells(GSE110509) or the exosomes derived from urine. QPCR confirmed that the expression of miR-223-3p was higher in urinary exosomes derived from kidney stone patients.
Conclusions: The expression of miR-223-3p is significantly higher in urinary exosomes from kidney stone patients and may affect stone formation via regulating oxidative stress, focal adhesion and inflammation process.
Source of Funding: None, there is no funding for these project
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