Abstract
Exosomes transmit certain amounts of molecules to specific recipient cells for intercellular communication. Among these molecules, messenger RNAs (mRNAs) may be delivered and translated into proteins in the recipient cells, and these mRNAs are thought to be critical mediators of exosomal functions. There are three subtypes of M2 macrophages (M2Ф), M2aФ, M2bФ, and M2cФ, which have different specific functional programs. The aim of the present study was to screen the mRNA profiles in the exosomes of these macrophage subtypes and to analyze the transcriptomic profile features associated with their specific functions. The mRNA contents of the exosomes isolated from the culture supernatants of the M2Ф subtypes were analyzed and compared using the Illumina HiSeq platform. The results indicated that the exosomes contained particular mRNAs from their source cells and were messengers of cellular functions. Bioinformatics analysis suggested that the exosomal mRNAs from M2bФs are enriched in the Toll-like receptor (TLR), tumor necrosis factor (TNF), NOD-like receptor (NLR), and NF-kappa B (NF-κB) signaling pathways. The mRNA profile of exosomes from M2bФ was distinctly different from that of exosomes from M2aФ and M2cФ and was consistent with the M2bФ cytological characteristic of maintaining a high level of proinflammatory cytokine and regulatory factor production. Therefore, the mRNA profiles revealed several characteristics of the exosomes from diverse forms of M2Ф. Further functional investigations based on these results may advance the understanding of the physiological roles of exosome-transferred mRNAs in MФ functions.
Highlights
Exosomes are small membrane-derived vesicles of endocytic origin that are secreted by most cells and are approximately 40–200 nm in diameter [1]
Both the Roswell Park Memorial Institute- (RPMI-)1640 and Dulbecco’s modified Eagle’s medium (DMEM) were supplemented with 10% fetal bovine serum (FBS; Gibco) and 10 ng/ml macrophage colony-stimulating factor (MCSF; PeproTech, Rocky Hill, NJ, USA)
The phenotypes of the polarized bone marrow-derived macrophages (BMDMs) were identified based on their gene expression using qPCR
Summary
Exosomes are small membrane-derived vesicles of endocytic origin that are secreted by most cells and are approximately 40–200 nm in diameter [1]. Exosomes are extracellular phospholipid nanocarriers that function as signalosomes and transmit molecules (DNA, RNA, proteins, and lipids) to specific recipient cells for intercellular communication [2, 3]. Among the molecules contained in exosomes, messenger RNAs (mRNAs) may be delivered and translated into proteins in recipient cells [3, 4]. Valadi et al [3] first reported that exosomes mediate the transfer of mRNAs and demonstrated that exosomes are capable of shuttling mRNAs between mast cells. Zomer et al [5] reported that exosomes are able to deliver the mRNA encoding Cre recombinase to recipient cells to perform Cre-LoxP-mediated recombination. MRNAs are thought to be critical mediators of exosomal functions
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