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J Extracell Vesicles. 2015 May 29;4:26760. doi: 10.3402/jev.v4.26760. eCollection 2015.

Quantitative and qualitative analysis of small RNAs in human endothelial cells and exosomes provides insights into localized RNA processing, degradation and sorting.

Author information

1
Department of Nephrology and Hypertension, UMC Utrecht, Utrecht, the Netherlands; b.w.m.vanbalkom@umcutrecht.nl.
2
Department of Nephrology and Hypertension, UMC Utrecht, Utrecht, the Netherlands.
3
Exosomes Research Group, VU University Medical Center, Amsterdam, the Netherlands.
4
ServiceXS B.V., Leiden, the Netherlands.

Abstract

Exosomes are small vesicles that mediate cell-cell communication. They contain proteins, lipids and RNA, and evidence is accumulating that these molecules are specifically sorted for release via exosomes. We recently showed that endothelial-cell-produced exosomes promote angiogenesis in vivo in a small RNA-dependent manner. Recent deep sequencing studies in exosomes from lymphocytic origin revealed a broad spectrum of small RNAs. However, selective depletion or incorporation of small RNA species into endothelial exosomes has not been studied extensively. With next generation sequencing, we identified all known non-coding RNA classes, including microRNAs (miRNAs), small nucleolar RNAs, yRNAs, vault RNAs, 5p and 3p fragments of miRNAs and miRNA-like fragments. In addition, we mapped many fragments of messenger RNAs (mRNAs) and mitochondrial RNAs (mtRNAs). The distribution of small RNAs in exosomes revealed a considerable overlap with the distribution in the producing cells. However, we identified a remarkable enrichment of yRNA fragments and mRNA degradation products in exosomes consistent with yRNAs having a role in degradation of structured and misfolded RNAs in close proximity to endosomes. We propose that endothelial endosomes selectively sequester cytoplasmic RNA-degrading machineries taking part in gene regulation. The release of these regulatory RNAs via exosomes may have implications for endothelial cell-cell communication.

KEYWORDS:

extracellular vesicles; microvesicles; next generation sequencing; quality control

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