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

• 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
• ISSN: 0022-0930; 1608-3202
• DOI: 10.1134/S0022093022060151

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.