Human multipotent mesenchymal stromal cells cytokine priming promotes RAB27B-regulated secretion of small extracellular
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(2020) 11:539
RESEARCH
Open Access
Human multipotent mesenchymal stromal cells cytokine priming promotes RAB27Bregulated secretion of small extracellular vesicles with immunomodulatory cargo Anastasia Cheng1, Dongsic Choi1, Maximilien Lora1, Dominique Shum-Tim2, Janusz Rak1 and Inés Colmegna1,3*
Abstract Background: The paracrine effects of multipotent mesenchymal stromal cells (MSCs) are mediated by their secretome composed by soluble factors (i.e., cytokines, growth factors, hormones) and extracellular vesicles (EVs). EVs promote intercellular communication, and the EV cargoes [e.g., proteins, soluble factors, microRNAs (miRNAs), messenger RNA (mRNA), DNA] reflect the molecular and functional characteristics of their parental cells. MSCderived EVs (MSC-EVs) are currently evaluated as subcellular therapeutics. A key function of the MSC secretome is its ability to promote immune tolerance (i.e., immunopotency), a property that is enhanced by priming approaches (e.g., cytokines, hypoxia, chemicals) and inversely correlates with the age of the MSC donors. We evaluated mechanisms underlying MSC vesiculation and the effects of inflammation and aging on this process. Methods: We evaluated the effects of interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α) on human adipose-derived MSC: (a) vesiculation (custom RT2 Profiler PCR Array), (b) EV profiles (Nanoparticle Tracking Analysis and Nanoparticle Flow Cytometry), (c) EV cargo (proteomic analysis and Western blot analysis), and (d) immunopotency (standard MSC:CD4 T cell proliferation inhibition assay). We confirmed the role of RAB27B on MSC vesiculation (RAB27B siRNA) and assessed its differential contribution to vesiculation in adult and pediatric MSCs (qPCR). Results: Cytokine priming upregulated RAB27B in adipose-derived MSCs increasing their secretion of exosome-like small EVs (sEVs; < 200 nm) containing two key mediators of immunopotency: A20 and TSG-6. These EVs inhibited T cell proliferation in a dose-dependent manner. RAB27B siRNA inhibited MSC vesiculation. Adipose-derived MSCs isolated from pediatric donors exhibited higher RAB27B expression and secreted more sEVs than adult MSCs. Conclusions: Cytokine priming is a useful strategy to harvest anti-inflammatory MSC-sEVs for clinical applications. Of relevance, donor age should be considered in the selection of MSC-sEVs for clinical applications. Keywords: Multipotent mesenchymal stromal cells, MSC, Extracellular vesicles, Exosomes, sEVs, RAB27B, TSG6, A20
* Correspondence: [email protected] 1 Research Institute of the McGill University Health Centre, McGill University, 1001 Decarie Blvd, Office # EM2-3238, Montreal, QC H4A 3J1, Canada 3 Division of Rheumatology, Department of Medicine, McGill University, Montreal, QC, Canada Full list of author information is available at the end of the article © The Author(s). 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in an
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