MicroRNA and mRNA cargo of extracellular vesicles from porcine adipose tissue-derived mesenchymal stem cells

Abstract

Mesenchymal stromal/stem cells (MSCs) are clinically useful for cell-based therapy, but concerns regarding their ability to replicate limit their human application. MSCs release extracellular vesicles (EVs) that mediate at least in part the paracrine effects of the parental cells. To understand the molecular basis of their biological properties, we characterized the RNA cargo of EVs from porcine adipose-tissue derived MSCs. Comprehensive characterization of mRNA and miRNA gene expression using high-throughput RNA sequencing (RNA-seq) revealed that EVs are selectively enriched for distinct classes of RNAs. For example, EVs preferentially express mRNA for transcription factors (e.g. MDFIC, POU3F1, NRIP1) and genes involved in angiogenesis (e.g. HGF, HES1, TCF4) and adipogenesis (e.g. CEBPA, KLF7). EVs also express Golgi apparatus genes (ARRB1, GOLGA4) and genes involved in TGF-β signaling. In contrast, mitochondrial, calcium signaling, and cytoskeleton genes are selectively excluded from EVs, possibly because these genes remain sequestered in organelles or intracellular compartments. RNA-seq generated reads for at least 386 annotated miRNAs, but only miR148a, miR532-5p, miR378, and let-7f were enriched in EVs compared to MSCs. Gene ontology analysis indicates that these miRNA target transcription factors and genes that participate in several cellular pathways, including angiogenesis, cellular transport, apoptosis, and proteolysis. Our data suggest that EVs transport gene regulatory information to modulate angiogenesis, adipogenesis, and other cell pathways in recipient cells. These observations may contribute to development of regenerative strategies using EVs to overcome potential complications of cell-based therapy.

Human Mesenchymal Stroma/Stem Cells Exchange Membrane Proteins and Alter Functionality During Interaction with Different Tumor Cell Lines

“…Exosomes represent the release of extracellular vesicles carrying predominantly proteins, mRNAs, and microRNAs that are exchanged during cellular interactions [57]. Production of exosomes has been described for MSC to transport gene regulatory information to recipient cells that can modulate cell growth and angiogenesis by affecting a variety of cellular pathways [58]. Indeed, the CD73, CD90, and CD105 induction in the different tumor cells was observed at both the protein and mRNA levels, suggesting exchange of proteins and/or transcriptional regulators and/or mRNAs between MSC and co-cultured tumor cells.…”

The metabolic syndrome alters the miRNA signature of porcine adipose tissue‐derived mesenchymal stem cells

“…We have previously shown that EVs released from porcine adipose tissue-derived MSCs transport gene regulatory information to modulate angiogenesis, adipogenesis, and other cell pathways in recipient cells [4, 5]. We have also shown that EVs are enriched with proteins linked to a broad range of biological functions, including angiogenesis, inflammation, apoptosis, and extracellular matrix remodeling[5] [6].…”

The secretome of mesenchymal stromal cells: Role of extracellular vesicles in immunomodulation

“…Moreover, EV-MSCs express some specific exosomal markers, such as CD107 [50,51], CD63, CD9 and CD81 [52,53].…”

“…Recently, the characterization of RNA content of EVs obtained from porcine ASCs has been reported [53]. EVs released from ASCs preferentially express mRNAs for transcription factors (MDFIC, POU3F1, NRIP1), genes involved in angiogenesis (HGF, HES1, TCF4) and adipogenesis (KLF7), Golgi apparatus genes (ARRB1, GOLGA4), and genes involved in TGF-␤ signaling.…”

“…Moreover, RNA-seq analyses generated reads for 386 miRNAs, but only 4 of them (miR148a, miR532-5p, miR378 and let-7f) are enriched in EVs with respect to ASCs, supporting the theory of a specific and organized package of miRNAs in EVs before their secretion. Gene ontology analyses have revealed that these miRNAs target transcription factors of genes involved in angiogenesis, cellular transport, apoptosis and proteolysis [53].…”