Urinary miRNA profiles discriminate between obstruction-induced bladder dysfunction and healthy controls

Michelle Von Siebenthal,Nadine Kunz,Akshay Akshay,Salomé Lizun-Platoni,Mustafa Besic,Ali Hashemi Gheinani,Katia Monastyrskaya,Fiona C Burkhard

doi:10.1038/s41598-021-89535-3

Abstract

Urgency, frequency and incomplete emptying are the troublesome symptoms often shared between benign prostatic obstruction-induced (BLUTD) and neurogenic (NLUTD) lower urinary tract dysfunction. Previously, using bladder biopsies, we suggested a panel of miRNA biomarkers for different functional phenotypes of the bladder. Urine is a good source of circulating miRNAs, but sex- and age-matched controls are important for urinary metabolite comparison. In two groups of healthy subjects (average age 32 and 57 years old, respectively) the total protein and RNA content was very similar between age groups, but the number of secreted extracellular vesicles (uEVs) and expression of several miRNAs were higher in the young healthy male volunteers. Timing of urine collection was not important for these parameters. We also evaluated the suitability of urinary miRNAs for non-invasive diagnosis of bladder outlet obstruction (BOO). A three urinary miRNA signature (miR-10a-5p, miR-301b-3p and miR-363-3p) could discriminate between controls and patients with LUTD (BLUTD and NLUTD). This panel of representative miRNAs can be further explored to develop a non-invasive diagnostic test for BOO. The age-related discrepancy in the urinary miRNA content observed in this study points to the importance of selecting appropriate, age-matched controls.

Highlights

Patients with lower urinary tract dysfunction (LUTD) suffer a range of symptoms such as increased daytime and nighttime frequency, urgency, urinary incontinence, slow stream, hesitancy and incomplete e mptying[1]
We propose a small panel of selected miRNAs, that can discriminate between patients with LUTD and controls
The samples from each time point were divided into 2 × 37 ml fractions and used for (1) total cell-free urinary RNA isolation and (2) serialcentrifugation (UC), protease inhibition, DL-dithiothreitol (DTT) treatment, filtration and size-exclusion chromatography (SEC) to isolate urinary extracellular vesicles (uEVs) for further a nalysis[27]

Summary

Introduction

Patients with lower urinary tract dysfunction (LUTD) suffer a range of symptoms such as increased daytime and nighttime frequency, urgency, urinary incontinence, slow stream, hesitancy and incomplete e mptying[1]. Urine is easy to collect, making it an attractive source of potential biomarkers It contains small molecule metabolites and a considerable number of proteins and nucleic acids, both free and packaged in urinary extracellular vesicles (uEVs)[8]. Having established an appropriate sample collection protocol, we compared the urinary miRNA profiles of the older control group with the samples from patients with benign prostatic obstruction-induced (BLUTD) and neurogenic (NLUTD) bladder dysfunction, and revealed significant changes in the miRNA content. We propose a small panel of selected miRNAs, that can discriminate between patients with LUTD and controls These selected miRNAs may upon further exploration have the potential for non-invasive urinary miRNA-based diagnosis of obstruction-induced bladder dysfunction

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