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Transcriptome profiling of macrophages exposed to extracellular vesicles containing miR-17 family.

ABSTRACT: Purpose: The goal of this study is to generate NGS-derived transcriptome profiles (RNA-seq) from bone marrow-derived macrophages (BMDMs) exposed to extracellular vesicles (EVs) containing the miR-17 family (miR-17 and miR-93). Methods: BMDMs were isolated from mouse bone and were exposed to lung epithelial Beas2B cell-derived EVs transfected with scrambled control RNAs, miR-17 mimics or miR-93 mimics. Whole transcriptome profiles of the exposed BMDMs (n = 4 biological replicates were pooled) were then generated using the Ion Proton™ System. Results: A wide range of transcript expressions were regulated by the EV-containing miR-17 and/or miR-93 in BMDMs. AC118255.2, Phox2a, 1700066J24Rik, and Gm20412 were effectively downregulated by both MV-miR-17 and MV-miR-93. Conclusions: Our study provides potential target transcripts of EV-containing miR-17/93 in macrophages.

ORGANISM(S): Mus musculus

PROVIDER: GSE129931 | GEO | 2019/04/18

REPOSITORIES: GEO

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Project description:Lifestyle disorders like obesity, type 2 diabetes (T2D), and cardiovascular diseases can be prevented and treated by regular physical activity. During exercise, skeletal muscles release signaling factors that communicate with other organs and mediate beneficial effects of exercise. These factors include myokines, metabolites, and extracellular vesicles (EVs). In the present study, we have examined how electrical pulse stimulation (EPS) of myotubes, a model of exercise, affects the cargo of released EVs. Chronic low frequency EPS was applied for 24 h to human myotubes isolated and differentiated from biopsy samples from 6 morbidly obese females with T2D, and EVs, both exosomes and microvesicles (MV), were isolated from cell media 24 h thereafter. Size and concentration of EV subtypes were characterized by nanoparticle tracking analysis, surface markers were examined by flow cytometry and Western blotting, and morphology was confirmed by transmission electron microscopy. Protein content was assessed by high-resolution proteomic analysis (LC-MS/MS), non-coding RNA was quantified by Affymetrix microarray, and selected microRNAs (miRs) validated by real time RT-qPCR. The size and concentration of exosomes and MV were unaffected by EPS. Of the 400 miRs identified in the EVs, EPS significantly changed the level of 15 exosome miRs, of which miR-1233-5p showed the highest fold change. The miR pattern of MV was unaffected by EPS. Totally, about 1000 proteins were identified in exosomes and 2000 in MV. EPS changed the content of 73 proteins in exosomes, 97 in MVs, and of these 4 were changed in both exosomes and MV (GANAB, HSPA9, CNDP2, and ATP5B). By matching the EPS-changed miRs and proteins in exosomes, 31 targets were identified, and among these several promising signaling factors. Of particular interest were CNDP2, an enzyme that generates the appetite regulatory metabolite Lac-Phe, and miR-4433b-3p, which targets CNDP2. Several of the regulated miRs, such as miR-92b-5p, miR-320b, and miR-1233-5p might also mediate interesting signaling functions.

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Transcriptome profiling of macrophages exposed to extracellular vesicles containing miR-17 family.

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