MiR-21-5p in urinary extracellular vesicles is a novel biomarker of urothelial carcinoma.

Kyosuke Matsuzaki,Yasushi Miyagawa,Motohide Uemura,Iwao Yoshioka,Kazutake Tsujikawa,Norio Nonomura,Rikinari Hanayama,Atsunari Kawashima,Kentaro Jingushi,Akira Nagahara,Kazutoshi Fujita,Go Tanigawa,Yuko Ueda,Koji Ueda,Takeshi Ujike

doi:10.18632/oncotarget.14969

Abstract

BackgroundExtracellular vesicles are lipid bilayer vesicles containing protein, messengerRNA and microRNA. Cancer cell-derived extracellular vesicles may be diagnostic and therapeutic targets. We extracted extracellular vesicles from urine of urothelial carcinoma patients and the control group to identify cancer-specific microRNAs in urinary extracellular vesicles as new biomarkers.Materials and methodsmicroRNA from urinary extracellular vesicles extracted from 6 urothelial carcinoma patients and 3 healthy volunteers was analyzed. We verified candidate microRNAs in an independent cohort of 60 urinary extracellular vesicles samples. To normalize the microRNA expression level in extracellular vesicles, we examined the following in extracellular vesicles: protein concentration, CD9 intensity, amounts of whole miRNAs, RNA U6B small nuclear expression and the creatinine concentration of original urine correlating with the counts of extracted extracellular vesicles measured by the NanoSight™ system.RESULTSFrom the microarray results 5 microRNAs overexpressed in urinary extracellular vesicles of urothelial carcinoma patients were identified. Creatinine concentration of original urine correlated most with particle counts of extracellular vesicles, indicating that creatinine could be a new tool for normalizing microRNA expression. MiR-21-5p was the most potent biomarker in urinary extracellular vesicles (sensitivity, 75.0%; specificity, 95.8%) and was also overexpressed in urinary extracellular vesicles from urothelial carcinoma patients with negative urine cytology. For the subgroup with negative urine cytology, the sensitivity was 75.0% and specificity was 95.8%.ConclusionMiR-21-5p in urinary extracellular vesicles could be a new biomarker of urothelial carcinoma, especially for urothelial carcinoma patients with negative urine cytology.

Highlights

Urothelial carcinomas (UC), including bladder cancer, are the sixth most common cancer in the United States [1]
To normalize the microRNA expression level in extracellular vesicles, we examined the following in extracellular vesicles: protein concentration, CD9 intensity, amounts of whole micro RNA (miRNA), RNA U6B small nuclear expression and the creatinine concentration of original urine correlating with the counts of extracted extracellular vesicles measured by the NanoSightTM system
Creatinine concentration of original urine correlated most with particle counts of extracellular vesicles, indicating that creatinine could be a new tool for normalizing microRNA expression

Summary

Introduction

Urothelial carcinomas (UC), including bladder cancer, are the sixth most common cancer in the United States [1]. The current standard of screening for UC is urine cytology, and its value in diagnosis UC is based on a sensitivity of 38.0% and a specificity of 98.3% [2]. Cystoscopy can offer high diagnostic accuracy, but it is costly and invasive, and in general, the urologists would perform cystoscopy only on the patients with a high level of suspicion of UC (e.g., positive urine cytology or gross hematuria). For these reasons, noninvasive and more sensitive biomarkers are needed. We extracted extracellular vesicles from urine of urothelial carcinoma patients and the control group to identify cancer-specific microRNAs in urinary extracellular vesicles as new biomarkers

Methods

Results

Conclusion