Small Non-Coding RNA Profiles of Sorted Plasma Extracellular Vesicles: Technical Approach

Extracellular vesicles play a pivotal role in the intercellular communications influencing various physiological and pathological processes. They carry a range of biomolecular cargoes including small non-coding RNAs (ncRNAs) that could serve as potential diagnostic biomarkers and therapeutic targets...

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Bibliographic Details
Published in:Journal of evolutionary biochemistry and physiology 2022-11, Vol.58 (6), p.1847-1864
Main Authors: Kalinina, O. V., Khudiakov, A. А., Panshin, D. D., Nikitin, Yu. V., Ivanov, A. M., Kostareva, A. A., Golovkin, A. S.
Format: Article
Language:English
Subjects:
Animal Physiology
Biochemistry
Biomedical and Life Sciences
CD45 antigen
Erythrocytes
Evolutionary Biology
Exosomes
Experimental Papers
Extracellular vesicles
Leukocytes
Life Sciences
MicroRNAs
miRNA
Next-generation sequencing
Non-coding RNA
Plasma
Platelets
RNA transport
Therapeutic targets
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Summary:Extracellular vesicles play a pivotal role in the intercellular communications influencing various physiological and pathological processes. They carry a range of biomolecular cargoes including small non-coding RNAs (ncRNAs) that could serve as potential diagnostic biomarkers and therapeutic targets. In the current study we applied Next Generation Sequencing to investigate small RNA profiles of erythrocytes, platelets, leukocytes, two plasma fractions and blood cell-derived extracellular vesicles (EVs) (CD41+, CD45+, CD235a+, CD146+) obtained using high-sensitivity fluorescence activated vesicle sorting (hs-FAVS) from plasma of healthy donors. We analyzed the proportions of various small ncRNAs across samples and identified sample specific profiles of microRNA (miRNA) and transport RNA-derived fragments (tRFs). It was found that the cumulative small ncRNA profiles of EVs originated from platelets, erythrocytes, and leukocytes, which are considered to be dominant among the vesicles circulating in blood, differ from small ncRNA profiles of plasma fractions, which represent macrovesicles and exosomes in the circulation. The proportion of miRNAs in sorted EVs was significantly lower compared to other samples while the proportion of tRFs was higher. Moreover, all sorted EVs carried mostly cell type non-specific miRNAs. Taken together, the results demonstrate that the combination of hs-FAVS with small-RNA sequencing technique is a powerful approach to analyze small ncRNA profiles in cell specific vesicles.
ISSN:0022-0930
1608-3202
DOI:10.1134/S0022093022060151