TY - JOUR
T1 - The profiles of microRNAs from urinary extracellular vesicles (EVs) prepared by various isolation methods and their correlation with serum EV microRNAs
AU - Park, Sungjin
AU - Lee, Kiyoung
AU - Park, Ie Byung
AU - Kim, Nan Hee
AU - Cho, Seongcheol
AU - Rhee, Won Jong
AU - Oh, Yujin
AU - Choi, Jimin
AU - Nam, Seungyoon
AU - Lee, Dae Ho
N1 - Funding Information:
Authors thank Curt Balch for editing this manuscript. DHL conceived design of the work, and SN and DHL supervised the work. SP, KL, IBP, NHK, SC, WJR, YO, JC, SN, DHL performed acquisition, analysis, or interpretation of data for the work. SP drafted the work and SN and DHL revised it. All authors have read and approved the manuscript.
Funding Information:
This work was supported by grants from the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Korea [grant numbers HI14C1135, HI16C1997].
Publisher Copyright:
© 2020
PY - 2020/2
Y1 - 2020/2
N2 - Aims: MicroRNAs (miRNAs) that circulate in biological fluids are frequently enclosed in extracellular vesicles (EVs). However, urinary EVs and their cargo miRNAs have not been systematically studied according to their EV isolation methods. Methods: In type 2 diabetes mellitus persons with diabetic nephropathy (n = 4), we compared miRNA species in urine EVs prepared by ultracentrifugation (UC), qEV original size exclusion column (qEV), ExoQuick-TC Plus (ExoQuick), and ultrafiltration using Amicon Ultra centrifugal filter devices (Amicons) 10 K and 100 K. EV miRNAs were profiled by next-generation sequencing (NGS). Additionally, we evaluated the correlations of EV miRNA expression between the urine and serum samples isolated by UC. Results: From each of 100 ml of urine, the UC method yielded the highest number of EV miRNA species (233 ± 37.3), with the ExoQuick yielded the lowest (103 ± 17.4). Urine EV miRNA profiles were highly correlated between UC, qEV, ExoQuick and Amicon 10 K methods. EV miRNA profiles between the urine and serum samples showed variable correlations between the patients (paired sample number = 3, r = 0.39–0.72). Conclusions: UC, qEV, ExoQuick, and Amicon 10 K are acceptable for urinary EV isolation to profile miRNAs. Urine- and serum-derived EV miRNA profiles have variable correlations depending on specific patients.
AB - Aims: MicroRNAs (miRNAs) that circulate in biological fluids are frequently enclosed in extracellular vesicles (EVs). However, urinary EVs and their cargo miRNAs have not been systematically studied according to their EV isolation methods. Methods: In type 2 diabetes mellitus persons with diabetic nephropathy (n = 4), we compared miRNA species in urine EVs prepared by ultracentrifugation (UC), qEV original size exclusion column (qEV), ExoQuick-TC Plus (ExoQuick), and ultrafiltration using Amicon Ultra centrifugal filter devices (Amicons) 10 K and 100 K. EV miRNAs were profiled by next-generation sequencing (NGS). Additionally, we evaluated the correlations of EV miRNA expression between the urine and serum samples isolated by UC. Results: From each of 100 ml of urine, the UC method yielded the highest number of EV miRNA species (233 ± 37.3), with the ExoQuick yielded the lowest (103 ± 17.4). Urine EV miRNA profiles were highly correlated between UC, qEV, ExoQuick and Amicon 10 K methods. EV miRNA profiles between the urine and serum samples showed variable correlations between the patients (paired sample number = 3, r = 0.39–0.72). Conclusions: UC, qEV, ExoQuick, and Amicon 10 K are acceptable for urinary EV isolation to profile miRNAs. Urine- and serum-derived EV miRNA profiles have variable correlations depending on specific patients.
KW - Biomarker
KW - Diabetic nephropathy
KW - Extracellular vesicle
KW - Serum
KW - Urine
KW - microRNA
UR - http://www.scopus.com/inward/record.url?scp=85078262819&partnerID=8YFLogxK
U2 - 10.1016/j.diabres.2020.108010
DO - 10.1016/j.diabres.2020.108010
M3 - Article
C2 - 31987752
AN - SCOPUS:85078262819
SN - 0168-8227
VL - 160
JO - Diabetes Research and Clinical Practice
JF - Diabetes Research and Clinical Practice
M1 - 108010
ER -
