Heterogeneity and interplay of the extracellular vesicle small RNA transcriptome and proteome

Sci Rep. 2018 Jul 17;8(1):10813. doi: 10.1038/s41598-018-28485-9.

Abstract

Extracellular vesicles (EVs) mediate cell-to-cell communication by delivering or displaying macromolecules to their recipient cells. While certain broad-spectrum EV effects reflect their protein cargo composition, others have been attributed to individual EV-loaded molecules such as specific miRNAs. In this work, we have investigated the contents of vesicular cargo using small RNA sequencing of cells and EVs from HEK293T, RD4, C2C12, Neuro2a and C17.2. The majority of RNA content in EVs (49-96%) corresponded to rRNA-, coding- and tRNA fragments, corroborating with our proteomic analysis of HEK293T and C2C12 EVs which showed an enrichment of ribosome and translation-related proteins. On the other hand, the overall proportion of vesicular small RNA was relatively low and variable (2-39%) and mostly comprised of miRNAs and sequences mapping to piRNA loci. Importantly, this is one of the few studies, which systematically links vesicular RNA and protein cargo of vesicles. Our data is particularly useful for future work in unravelling the biological mechanisms underlying vesicular RNA and protein sorting and serves as an important guide in developing EVs as carriers for RNA therapeutics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Chromatography, High Pressure Liquid
  • Extracellular Vesicles / metabolism*
  • HEK293 Cells
  • Humans
  • Proteome / analysis*
  • RNA, Small Interfering / chemistry
  • RNA, Small Interfering / metabolism
  • RNA, Small Nuclear / chemistry
  • RNA, Small Nuclear / metabolism*
  • Sequence Analysis, RNA
  • Tandem Mass Spectrometry
  • Transcriptome*

Substances

  • Proteome
  • RNA, Small Interfering
  • RNA, Small Nuclear