Adipose derived mesenchymal stem cell exosomes loaded with miR-10a promote the differentiation of Th17 and Treg from naive CD4+ T cell
Introduction
CD4+ T cells have a central role in orchestrating the immune system. Various subsets of CD4+ T cells including regulatory (Treg) cells and effector T helper (Th) cells, especially Th1, Th2 and Th17, should work in equilibrium to ensure proper immune response against foreign pathogens and cancer [1,2]. Imbalance between CD4+ T cell subsets leads to dysregulated immune responses such as immune tolerance to pathogens and tumors or autoimmune/inflammatory diseases. In particular, the imbalance between Th17 and Treg is emerging as the major etiologic factor in autoimmunity and cancer [3,4].
To ensure the balance between various subsets of CD4+ T cells, their differentiation from naïve CD4+ T cells is precisely regulated by multifaceted interplay of several cytokines and microRNAs which activate subset-specific transcription factors [4,5]. In this regard, exploring the factors involved in the differentiation of naive T cells to CD4+ T cell subsets, as well as improving the methods for fine tuning subset specific responses, have great clinical importance. Emergence of adoptive T cell transfer therapies has further increased the significance of precise exvivo control over differentiation of naïve T cells [4].
Several microRNAs have been so far revealed as regulators of the balance between CD4+ T cell subsets [6]. In particular, miR-10a has been reported to be preferentially expressed in naturally occurring Tregs [12,14], showing an inverse correlation with susceptibility to autoimmune disease [7,8]. In the presence of retinoic acid, retroviral overexpression of miR-10a in mouse naïve CD4+ T cells has been shown to inhibit their differentiation to Th17 [9]. In addition, lentiviral transduction of miR-10a in CD4+ T cells from human peripheral blood has been reported to suppress Th1 and Th17 responses without affecting those of Th2 and Treg [10]. However, the impacts of nonvirally transfected miR-10a on CD4+ T cell subsets have not been studied yet.
Mesenchymal stem cells (MSCs) are well known for their immunomodulatory roles, promising a wide range of therapeutic potentials in immune-related disorders [11]. The immunomodulation exerted by MSCs is substantially mediated through regulating the fate of T cells to maintain the balance between CD4+ T cell subsets [12]. The secretome of MSCs, especially the MSC derived exosomes, have the major role in transmitting the regulatory signals to immune cells [13]. In addition to their therapeutic potentials, MSC derived exosomes are emerging as ideal vehicles for delivery of small non-coding RNAs to target cells [14,15]. Notably, MSC derived exosomes are highly enriched in miR-10a, which might further enhance their efficacy as vehicles for delivery of this microRNA [[16], [17], [18]].
Along with our studies on therapeutic potentials of noncoding RNAs delivered by MSC derived exosomes, in this study we examined the impacts of miR-10a loaded MSC derived exosomes on differentiation of naive CD4+ T cells. For this purpose, human adipose tissue derived mesenchymal stem cells (AD-MSCs) as the most advantageous source of MSCs were isolated and their secreted exosomes (AD-MSC-Exo) were transfected with miR-10a. Following treatment with miR-10a loaded AD-MSC-Exos, in vitro differentiation of mouse naïve CD4+ T cells toward Th1, Th2, Th17, and Treg was examined by measuring the expression levels of subset-specific cytokines and transcription factors.
Section snippets
Isolation of AD-MSCs
The study design is schematically represented in Fig. 1. AD-MSCs were isolated from adipose tissue of 3 healthy adult donors as previously described [19]. In brief, samples of adipose tissue were enzymatically dissociated at 37 °C with type I collagenase (0.075%) for 20 min, centrifuged at 500g for 10 min and the pellets were resuspened in Dulbecco's modified Eagle medium (DMEM) with 10% Fetal Bovine Serum (FBS) (All from Gibco, Waltham, MA, USA) and 1% pen/strep and incubated at 37 °C in 5%
Characterization of AD-MSCs
MSCs were purified from human adipose tissue and characterized via morphological properties and differentiation to adipocytes and osteocytes at passage 3. The cells displayed a spindle-like morphology (Fig. 2A). In order to determine the differentiation potency of isolated cells, after 21 days incubation with osteogenic and adipogenic media, their differentiation was evaluated with Alizarin red S and Oil Red O staining, respectively (Fig. 2B and C). Furthermore, the purity of MSCs was
Discussion
miR-10a is known as a regulator of CD4+ T cell fate according to former reports based on viral transduction [[7], [8], [9], [10]]. Considering AD-MSC-Exos as effective immunomodulators as well as advantageous vehicles for delivery of microRNAs [13], the hypothetical regulatory roles of miR-10a on the fate of CD4+ T cells need to be examined using MSC-Exo mediated delivery. In this study, as a proof of the concept, we have shown effective induction of miR-10a in naïve CD4+ T cells using
Conclusion
Recent studies have highlighted the roles of Th17 in antitumor immunity [4]. Accordingly, novel methods are under investigation to provide Th17 dominant responses including the exvivo expansion of Th17 cells to be used as adoptive transfer therapy. In novel anti-tumor immunotherapeutic strategies, Tregs and Th17 are used in concert to maximize the efficiency against tumor while minimizing the toxicity against self tissue. miR-10a loaded AD-MSC-Exos, concomitantly promoting both Treg and Th17
Declaration of competing interest
The authors declare that there are neither ethical nor financial conflicts of interest involved in the manuscript.
Acknowledgments
This work was supported by the Shahid Beheshti University of Medical Sciences (Grant No: 6604) and the Iranian Stem Cell Council, Tehran, Iran.
References (29)
- et al.
Th17 and regulatory T cell balance in autoimmune and inflammatory diseases
Autoimmun. Rev.
(2014) - et al.
Phenotypical and functional evaluation of dendritic cells after exosomal delivery of miRNA-155
Life Sci.
(2019) - et al.
In vitro analysis of immunomodulatory effects of mesenchymal stem cell- and tumor cell-derived exosomes on recall antigen-specific responses
Int. Immunopharmacol.
(2019) - et al.
MSC-derived extracellular vesicles attenuate immune responses in two autoimmune murine models: type 1 diabetes and uveoretinitis
Stem Cell Rep
(2017) - et al.
Mesenchymal stem cell exosomes promote immunosuppression of regulatory T cells in asthma
Exp. Cell Res.
(2018) - et al.
Differentiation and function of T cell subsets in infectious diseases
J Immunol Res
(2018) Regulators of T-cell fate: integration of cell migration, differentiation and function
Immunol. Rev.
(2019)- et al.
When worlds collide: Th17 and Treg cells in cancer and autoimmunity
Cell. Mol. Immunol
(2018) - et al.
The multifaceted Interface between cytokines and microRNAs: an ancient mechanism to regulate the good and the bad of inflammation
Front. Immunol.
(2018) - et al.
MicroRNA-mediated regulation of T helper type 17/regulatory T-cell balance in autoimmune disease
Immunology
(2018)
MicroRNA 10a marks regulatory T cells
PLoS One
miR-182 and miR-10a are key regulators of Treg specialisation and stability during Schistosome and Leishmania-associated inflammation
PLoS Pathog.
TGF-β and retinoic acid induce the microRNA miR-10a, which targets Bcl-6 and constrains the plasticity of helper T cells
Nat. Immunol.
miR-10a inhibits dendritic cell activation and Th1/Th17 cell immune responses in IBD
Gut
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