Elsevier

Journal of Autoimmunity

Volume 70, June 2016, Pages 31-39
Journal of Autoimmunity

Human adipose mesenchymal stem cells as potent anti-fibrosis therapy for systemic sclerosis

https://doi.org/10.1016/j.jaut.2016.03.013Get rights and content

Highlights

  • Mesenchymal stem cells (MSC) are promising in treating systemic sclerosis (SSc).

  • Allogeneic MSC reduce skin and lung fibrosis in the HOCl-model of SSc.

  • Beyond species barrier, human MSC efficiently reduce tissue fibrosis in this model.

  • Human adipose MSC exert more potent effects than bone-marrow MSC.

  • Adipose tissue should be considered as a source of MSC in future clinical trials.

Abstract

Objectives

Displaying immunosuppressive and trophic properties, mesenchymal stem/stromal cells (MSC) are being evaluated as promising therapeutic options in a variety of autoimmune and degenerative diseases. Although benefits may be expected in systemic sclerosis (SSc), a rare autoimmune disease with fibrosis-related mortality, MSC have yet to be evaluated in this specific condition. While autologous approaches could be inappropriate because of functional alterations in MSC from patients, the objective of the present study was to evaluate allogeneic and xenogeneic MSC in the HOCl-induced model of diffuse SSc. We also questioned the source of human MSC and compared bone marrow- (hBM-MSC) and adipose-derived MSC (hASC).

Methods

HOCl-challenged BALB/c mice received intravenous injection of BM-MSC from syngeneic BALB/c or allogeneic C57BL/6 mice, and xenogeneic hBM-MSC or hASC (3 donors each). Skin thickness was measured during the experiment. At euthanasia, histology, immunostaining, collagen determination and RT-qPCR were performed in skin and lungs.

Results

Xenogeneic hBM-MSC were as effective as allogeneic or syngeneic BM-MSC in decreasing skin thickness, expression of Col1, Col3, α-Sma transcripts, and collagen content in skin and lungs. This anti-fibrotic effect was not associated with MSC migration to injured skin or with long-term MSC survival. Interestingly, compared with hBM-MSC, hASC were significantly more efficient in reducing skin fibrosis, which was related to a stronger reduction of TNFα, IL1β, and enhanced ratio of Mmp1/Timp1 in skin and lung tissues.

Conclusions

Using primary cells isolated from 3 murine and 6 human individuals, this preclinical study demonstrated similar therapeutic effects using allogeneic or xenogeneic BM-MSC while ASC exerted potent anti-inflammatory and remodeling properties. This sets the proof-of-concept prompting to evaluate the therapeutic efficacy of allogeneic ASC in SSc patients.

Introduction

Systemic sclerosis (SSc) is an orphan disease characterized by tissue fibrosis, microangiopathy and autoimmunity, still exhibiting poor prognosis in many patients [1]. One of the most current promising therapeutic approaches is cell therapy, including hematopoietic stem cell [2] and mesenchymal stem/stromal cell (MSC) transplantation [3], [4]. MSC are multipotent stromal progenitor cells that can be isolated from numerous tissues including bone marrow (BM), adipose tissue, synovium, dental pulp, umbilical cord, etc. They display immunomodulatory and trophic properties, among which their anti-fibrotic capacity is well described [5]. MSC have proven efficacy in several animal models of fibrosis [6], [7], [8] and we recently demonstrated in a murine model of HOCl-induced SSc, that an infusion of murine syngeneic MSC could alleviate skin and lung fibrosis through the modulation of inflammation, oxidative status and extracellular matrix remodeling [9].

While MSC-based clinical trials enrolling patients in phase I/II studies are ongoing, the finding of SSc-related alterations of MSC in their niche is of importance [10], [11], [12], [13], [14]. The question of using an allogeneic rather than autologous approach is therefore under debate. Another important issue regarding MSC-based therapy concerns the tissue source from which the cells are to be isolated. The most commonly used source of MSC is BM but an increasing number of studies investigate the potential of MSC isolated from subcutaneous fat, for obvious easier accessibility and higher recovery yield [15]. BM-derived MSC (BM-MSC) and adipose-derived MSC (ASC) share a common phenotype, differentiation potential and trophic function but exhibit disparities in the range of their functional and therapeutic activity [16], [17], [18], [19]. Moreover, the different MSC sources have scarcely been compared in preclinical or clinical studies [20], [21], [22], [23] and never investigated in the specific conditions of SSc.

We therefore evaluated the therapeutic potential of BM-MSC according to antigen compatibility and compared the efficacy of allogeneic and xenogeneic BM-MSC versus autologous/syngeneic BM-MSC in the murine preclinical model of HOCl-induced diffuse SSc. In this model, we also investigated the therapeutic effect of human ASC, obtained from several donors, by comparison with human BM-MSC.

Section snippets

Experimental design and animals

SSc was induced by daily intradermal injections of hypochlorite (HOCl) as previously described [9], [24] and according to the Laboratory Animal Care guidelines with approval from the Regional Ethics Committee on Animal Experimentation (CEEA-LR-11054). A healthy control group was injected with phosphate buffered saline (PBS). All experiments were performed in BALB/c mice, except for the biodistribution study performed in C57BL/6 mice. At day 21, homogeneous HOCl-challenged groups of mice were

Isolation and characterization of MSC

Murine BM-MSC (mBM-MSC) were isolated from BALB/c and C57BL/6 mice as previously described and further characterized [9]. mBM-MSC were made of a homogeneous population of cells that expressed the conventional markers for stromal progenitors Sca-1, CD29, CD44 and did not express the hematopoietic markers CD11b, CD45 or F4/80 (Fig. 1A). Human MSC isolated from BM (hBM-MSC) or adipose tissue (hASC) highly expressed the stromal progenitor markers CD73, CD90, CD13, and CD105 (Fig. 1A). Both cell

Discussion

In the last decades, MSC have been shown to exert potent immunosuppressive properties, affecting both the innate and adaptive immune responses, through the inhibition of immune cell proliferation and differentiation, and the promotion of immune tolerance by the generation of regulatory cells [28]. Hence, there has been a rising interest for MSC-based therapy in the field of autoimmune diseases, with promising results in various animal models and phase I/II clinical trials in multiple sclerosis,

Conclusion

To conclude, this preclinical study demonstrated that, beyond major histocompatibility antigen mismatch, MSC-based therapy still remained efficient in reducing skin and lung fibrosis in murine SSc, which is promising concerning allogeneic approaches in the human refractory disease. The potent effect obtained with human ASC underlined the interest of using subcutaneous adipose tissue rather than BM as a source of MSC in future clinical trials. However, the wide clinical heterogeneity of the

Competing interests

ALQ declares speaking fees from Actelion Pharmaceuticals. None of the other authors has any potential conflict of interest related to this manuscript.

Acknowledgments

Work in the laboratory Inserm U1183 was supported by the Inserm Institute and the University of Montpellier. We are indebted to Montpellier-Nîmes University Hospital and Association des Sclérodermiques de France (ASF) for funding. ATJM received a fellowship from French Health ministry and Inserm institute for this work. We acknowledge the Agence Nationale pour la Recherche for support of the national infrastructure: “ECELLFRANCE: Development of a national adult mesenchymal stem cell based

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