Preanalytical considerations in quantifying circulating miRNAs that predict end-stage kidney disease in diabetes.

Publication Title

JCI Insight

Authors

Eiichiro Satake
Bozena Krolewski
Hiroki Kobayashi
Zaipul I Md Dom
Joseph Ricca
Jonathan M Wilson
Dave Hoon, Department of Translational Molecular Medicine, Saint John's Cancer Institute, Providence Health and Service, Santa Monica, California, USA.Follow
Kevin L Duffin
Marcus G Pezzolesi
Andrzej S Krolewski

Document Type

Article

Publication Date

6-24-2024

Keywords

californai; sjci; Humans; Circulating MicroRNA; Kidney Failure, Chronic; Male; Female; Middle Aged; Diabetic Nephropathies; Biomarkers; Aged; Reproducibility of Results; Adult; MicroRNAs; Disease Progression; Diabetes Mellitus

Abstract

Our previous study identified 8 risk and 9 protective plasma miRNAs associated with progression to end-stage kidney disease (ESKD) in diabetes. This study aimed to elucidate preanalytical factors that influence the quantification of circulating miRNAs. Using the EdgeSeq platform, which quantifies 2,002 miRNAs in plasma, including ESKD-associated miRNAs, we compared miRNA profiles in whole plasma versus miRNA profiles in RNA extracted from the same plasma specimens. Less than half of the miRNAs were detected in standard RNA extraction from plasma. Detection of individual and concentrations of miRNAs were much lower when RNA extracted from plasma was quantified by RNA sequencing (RNA-Seq) or quantitative reverse transcription PCR (qRT-PCR) platforms compared with EdgeSeq. Plasma profiles of miRNAs determined by the EdgeSeq platform had excellent reproducibility in assessment and had no variation with age, sex, hemoglobin A1c, BMI, and cryostorage time. The risk ESKD-associated miRNAs were detected and measured accurately only in whole plasma and using the EdgeSeq platform. Protective ESKD-associated miRNAs were detected by all platforms except qRT-PCR; however, correlations among concentrations obtained with different platforms were weak or nonexistent. In conclusion, preanalytical factors have a profound effect on detection and quantification of circulating miRNAs in ESKD in diabetes. Quantification of miRNAs in whole plasma and using the EdgeSeq platform may be the preferable method to study profiles of circulating cell-free miRNAs associated with ESKD and possibly other diseases.

Area of Special Interest

Kidney & Diabetes

Specialty/Research Institute

Endocrinology

Specialty/Research Institute

Nephrology

DOI

10.1172/jci.insight.174153