Small non-coding RNA's profile of sorted plasma extracellular vesicles expressing different cell origin

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 which 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 (Rbc), platelets (Plt), leukocytes (CD45cells), two plasma fractions and blood cell-derived extra-cellular vesicles (EVs, such as CD41+, CD45+, CD235a+, CD146+) obtained using high-sensitivity fluorescence activated vesicle sorting 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 ncRNAs profiles of EVs originated from platelets, erythrocytes, and leukocytes, which are considered to be dominant among the vesicles circulating in blood, differ from small ncRNAs profiles of plasma fractions, which represent macrovesicles and exosomes in 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. Taking together, the results demonstrate that the combination of high-sensitivity fluorescence activated vesicle sorting with small-RNA sequencing technique is a powerful approach to analyze small ncRNAs profiles in cell specific vesicles. Overall design: Comparative small RNA profiling of sorted plasma extracellular vesicles (CD41+, CD45+, CD235a+, CD146+), plasma fractions and blood cells (erythrocytes, platelets, leukocytes) from healthy donors