Extracellular microRNAs exhibit sequence-dependent stability and cellular release kinetics

Multiple studies have described extracellular microRNAs (ex-miRNAs) as being remarkably stable despite the hostile extracellular environment, when stored at 4ºC or lower. Here we show that many ex-miRNAs are rapidly degraded when incubated at 37ºC in the presence of serum (thereby simulating physiol...

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Bibliographic Details
Published in:RNA biology 2019-05, Vol.16 (5), p.696-706
Main Authors: Coenen-Stass, Anna M. L., Pauwels, Marie J., Hanson, Britt, Martin Perez, Carla, Conceição, Mariana, Wood, Matthew J. A., Mäger, Imre, Roberts, Thomas C.
Format: Article
Language:English
Subjects:
biomarkers
blood serum
cell culture
culture media
ex-miRNA
Extracellular microRNA
gel chromatography
genes
half life
kinetics
mathematical models
mice
microRNA
miRNA
myotubes
Research Paper
serum
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Summary:Multiple studies have described extracellular microRNAs (ex-miRNAs) as being remarkably stable despite the hostile extracellular environment, when stored at 4ºC or lower. Here we show that many ex-miRNAs are rapidly degraded when incubated at 37ºC in the presence of serum (thereby simulating physiologically relevant conditions). Stability varied widely between miRNAs, with half-lives ranging from ~1.5 hours to more than 13 hours. Notably, ex-miRNA half-lives calculated in two different biofluids (murine serum and C2C12 mouse myotube conditioned medium) were highly similar, suggesting that intrinsic sequence properties are a determining factor in miRNA stability. By contrast, ex-miRNAs associated with extracellular vesicles (isolated by size exclusion chromatography) were highly stable. The release of ex-miRNAs from C2C12 myotubes was measured over time, and mathematical modelling revealed miRNA-specific release kinetics. While some ex-miRNAs reached the steady state in cell culture medium within 24 hours, the extracellular level of miR-16 did not reach equilibrium, even after 3 days in culture. These findings are indicative of miRNA-specific release and degradation kinetics with implications for the utility of ex-miRNAs as biomarkers, and for the potential of ex-miRNAs to transfer gene regulatory information between cells.
ISSN:1547-6286
1555-8584
1555-8584
DOI:10.1080/15476286.2019.1582956