Whole blood holding time prior to plasma processing alters microRNA expression profile
File(s)
Published version
Author(s)
Type
Journal Article
Abstract
MicroRNAs (miRNAs) can exhibit aberrant expression under different physiological and pathological conditions. Therefore, differentially expressed circulating miRNAs have been a focus of biomarker discovery research. However, the use of circulating miRNAs comes with challenges which may hinder the reliability for their clinical application. These include varied sample collection protocols, storage times/conditions, sample processing and analysis methods. This study focused on examining the effect of whole blood holding time on the stability of plasma miRNA expression profiles. Whole blood samples were collected from healthy pregnant women and were held at 4°C for 30 min, 2 h, 6 h or 24 h prior to processing for plasma isolation. Plasma RNA was extracted and the expression of 179 miRNAs were analyzed. Unsupervised principal component analysis demonstrated that whole blood holding time was a major source of variation in miRNA expression profiles with 53 of 179 miRNAs showing significant changes in expression. Levels of specific miRNAs previously reported to be associated with pregnancy-associated complications such as hsa-miR-150-5p, hsa-miR-191-5p, and hsa-miR-29a-3p, as well as commonly used endogenous miRNA controls, hsa-miR-16-5p, hsa-miR-25-3p, and hsa-miR-223-3p were significantly altered with increase in blood holding time. Current protocols for plasma-based miRNA profiling for diagnostics describe major differences in whole blood holding periods ranging from immediately after collection to 26 h after. Our results demonstrate holding time can have dramatic effects on analytical reliability and reproducibility. This highlights the importance of standardization of blood holding time prior to processing for plasma in order to minimize introduction of non-biological variance in miRNA profiles.
Date Issued
2022-01-14
Date Acceptance
2021-12-21
Citation
Frontiers in Genetics, 2022, 12
ISSN
1664-8021
Publisher
Frontiers Media
Journal / Book Title
Frontiers in Genetics
Volume
12
Copyright Statement
© 2022 Kim, MacIntyre, Sykes, Arianoglou, Bennett and Terzidou. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
License URL
Sponsor
March of Dimes
Identifier
https://www.frontiersin.org/articles/10.3389/fgene.2021.818334/full
Grant Number
22-FY18-821
Subjects
Science & Technology
Life Sciences & Biomedicine
Genetics & Heredity
miRNA
microRNA
plasma
biomarker
hemolysis
whole blood
CIRCULATING MICRORNAS
SERUM
BIOMARKER
MIR-29A
CELLS
biomarker
hemolysis
miRNA - microRNA
plasma
whole blood
0604 Genetics
1103 Clinical Sciences
1801 Law
Publication Status
Published online
Date Publish Online
2022-01-14