Poster - #VH-42544 - Keywords: Anti-coagulant Extracellular Vesicles miRNA

Choice of Blood Collection Anticoagulant Influences Extracellular Vesicle and miRNA Profiling

Vivian Tran , Getulio Pereira de Oliveira Jr , Stephanie Chidester , Shulin Lu , Michelle L Pleet , Alexander R Ivanov , John Tigges , Moua Yang , ​ Steven Jacobson , Maria C. B. Gonçalves , Alec A. Schmaier , Jennifer Jones , Ionita C. Ghiran​
Drexel University College of Medicine, Beth Israel Deaconess Medical Center, Northeastern University, National Institutes of Health

ABSTRACT:
Circulating RNAs have been investigated systematically, both as constituents of circulating extracellular vesicles (EVs) and as non-EV RNA carriers. The high level of variability and low reproducibility rate of EV/extracellular RNA (exRNA) results promoted several efforts to limit pre-analytical variability by standardizing sample collection and preparation, instrument validation, setup and calibration. In this study, the focus was on the effect of anticoagulants (ACs) on the circulating EVs’ properties and on a set of circulating miRNA species, which were shown to be relevant disease markers. While the number of plasma EVs, their relative size, and post-fluorescence labeling profile varied with each AC, their overall immunological antigenic composition, with the exception of serum EVs, did not change significantly. For miRNA analysis, increase of expression levels in EDTA compared to serum may be associated with post-collection cellular stimulation and was not indicative of the circulating miRNA populations. Differences observed from each AC suggests that pre-study investigation of ACs are critical for obtaining data that better reflect the in vivo environment.