Characterization of mRNA and microRNA in human mast cell‐derived exosomes and their transfer to other mast cells and blood CD34 progenitor cells

Abstract

BackgroundExosomes are nanosized vesicles of endocytic origin that are released into the extracellular environment by many different cells. It has been shown that exosomes from various cellular origins contain a substantial amount of RNA (mainly mRNA and microRNA). More importantly, exosomes are capable of delivering their RNA content to target cells, which is a novel way of cell-to-cell communication. The aim of this study was to evaluate whether exosomal shuttle RNA could play a role in the communication between human mast cells and between human mast cells and human CD34+ progenitor cells.MethodsThe mRNA and microRNA content of exosomes from a human mast cell line, HMC-1, was analysed by using microarray technology. Co-culture experiments followed by flow cytometry analysis and confocal microscopy as well as radioactive labeling experiments were performed to examine the uptake of these exosomes and the shuttle of the RNA to other mast cells and CD34+ progenitor cells.ResultsIn this study, we show that human mast cells release RNA-containing exosomes, with the capacity to shuttle RNA between cells. Interestingly, by using microRNA microarray analysis, 116 microRNAs could be identified in the exosomes and 134 microRNAs in the donor mast cells. Furthermore, DNA microarray experiments revealed the presence of approximately 1800 mRNAs in the exosomes, which represent 15% of the donor cell mRNA content. In addition, transfer experiments revealed that exosomes can shuttle RNA between human mast cells and to CD34+ hematopoietic progenitor cells.ConclusionThese findings suggest that exosomal shuttle RNA (esRNA) can play a role in the communication between cells, including mast cells and CD34+ progenitor cells, implying a role in cells maturation process.

Human Mast Cell Sensitization with IgE Increases miRNA-210 Expression

“…It is known that miR-210 is upregulated during hypoxia, stimulates angiogenesis, induces proliferation, and decreases apoptosis [9]. In an allergic context, miR-210 expression has been shown to increase in mouse lymph nodes and T regulatory cells following dermal chemical sensitization [10], and miR-210 is upregulated in exosomes released by the HMC-1 cell line [11]. …”

Emerging roles of exosomes during epithelial–mesenchymal transition and cancer progression

“…Exosomes containing mRNAs can be transferred and influence the translational profile of recipient cells and tumour progression [47,72,76,77]. For example, Skog et al showed that by incorporating mRNA for a reporter oncogenic protein EGFRvIII into EVs, these transcripts (3426 transcripts present at different levels in exosomes and their donor cells) could be delivered to, and translated by recipient cells [77].…”

Mast Cells, Basophils and Mucosal Immunity

“…Exosomes contain a cornucopia of lipids, proteins, mRNA, and microRNA species and represent a novel mechanism of cell-cell communications, including exchange of genetic information (van Niel et al, 2006;Record et al, 2011). Mast cell exosomes can directly activate other cells (Skokos et al, 2001;Al-Nedawi et al, 2005), transfer mRNA and selective species of microRNA to other cells (Ekstrom et al, 2012), as well as several proteins (Valadi et al, 2007). Mast cell exosomes released during stress responses may be able to protect other cells from similar injury (Eldh et al, 2010), an effect likely mediated by transferred RNA.…”