Mesenchymal Stem Cells in Systemic Sclerosis: Allogenic or Autologous Approaches for Therapeutic Use?

Systemic sclerosis (SSc) is a rare autoimmune disease, which is potentially lethal. The physiopathology of the disease is still incompletely elucidated although the role of fibroblasts, endothelial cells (ECs), immune cells. and the environment (i.e., oxidative stress) has been demonstrated. This is an intractable disease with an urgent need to provide better therapeutic options to patients. Mesenchymal stem cells (MSCs) represent a promising therapeutic approach thanks to the number of trophic and pleiotropic properties they exert. Among these, MSCs display anti-fibrotic, angiogenic, and immunomodulatory capacities that might be of interest in the treatment of SSc by acting on different processes that are dysregulated in the disease. In the recent years, the therapeutic effectiveness of MSCs has been demonstrated in different preclinical animal models and is being investigated in phase I clinical trials. Both allogenic and autologous transplantation of MSCs isolated from bone marrow or adipose tissue is being evaluated. The rationale for using allogenic MSCs in SSc, as well as in other autoimmune diseases, is based on the possibility that autologous MSCs might be altered in these diseases. In SSc, reports from the literature are controversial. Nevertheless, the role of the oxidative environment and of the crosstalk with neighboring cells (fibroblasts and ECs) on the functional properties of MSCs has been reported. Here, we review the preclinical and clinical data reporting the interest of MSC-based treatment in SSc and question the use of autologous or allogeneic MSCs in perspective of clinical applications

Intriguing Relationships Between Cancer and Systemic Sclerosis: Role of the Immune System and Other Contributors

Systemic sclerosis (SSc) is an autoimmune connective tissue disorder, characterized by multisystem involvement, vasculopathy, and fibrosis. An increased risk of malignancy is observed in SSc (including breast and lung cancers), and in a subgroup of patients with specific autoantibodies (i.e., anti-RNA polymerase III and related autoantibodies), SSc could be a paraneoplastic syndrome and might be directly related to an immune response against cancer. Herein, we reviewed the literature, focusing on the most recent articles, and shed light onto the potential relationship between cancer and scleroderma regarding temporal and immunological dimensions

Therapeutic effect of extracellular vesicles derived from mesenchymal stromal cells in systemic sclerosis

La sclérodermie systémique (ScS) est une maladie rare et sévère, caractérisée par une dérégulation des cellules immunitaires, une fibrose et une vasculopathie généralisées. Aucun traitement curatif n’est validé à ce jour. Les cellules stromales mésenchymateuses (CSMs), dont l’efficacité a été démontrée dans plusieurs modèles précliniques de ScS, sont actuellement évaluées dans des essais cliniques. Elles exercent leurs propriétés régénératrices au travers de la sécrétion de médiateurs libérés dans le milieu extracellulaire ou contenus dans des vésicules extracellulaires (EVs). Nous nous intéressons ici au rôle des EVs libérées par les CSMs dans leur effet systémique au cours de la ScS.Dans un modèle murin de ScS induit par des injections d’acide hypochloreux, nous démontrons que les EVs stoppent l’évolution de la maladie au niveau cutané et pulmonaire sur le plan clinique, histologique et moléculaire. Nous identifions miR-29a-3p au sein des CSMs et de leurs EVs et prouvons qu’il est garant de leur effet thérapeutique, en ciblant Dnmt3a et Pdgfr-bb, deux nouvelles cibles identifiées dans la ScS.Nous améliorons l’efficacité thérapeutique des EVs sur les anomalies pulmonaires en pré-activant les CSMs à l’aide d’IFNγ alors que l’augmentation de la quantité d’EVs injectées n’a aucun effet. L’IFNγ module l’expression de molécules immunosuppressives dans les CSMs et leurs EVs, potentialisant ainsi leurs propriétés thérapeutiques.Alors que le rôle des CSMs de patients (CSMs-ScS) dans le développement de la maladie est sujet à débat, nous montrons que les CSMs-ScS inhibent in vitro la prolifération des lymphocytes T de manière similaire à des CSMs saines. En outre, elles sont capables d’améliorer le phénotype myofibroblastique de fibroblastes stimulés par du TGFβ1 qui présentent un comportement similaire à celui de fibroblastes de patients dans le modèle in vitro mis en place.En conclusion, les EVs sont aussi efficaces que les CSMs pour traiter la ScS murine et le pré-conditionnement des CSMs par l’IFNγ améliore leur effet au niveau pulmonaire. Les CSMs de patients conservent leurs fonctions anti-fibrotique et immunosuppressive in vitro et une de nos perspectives est d’évaluer leur efficacité thérapeutique in vivo.Systemic sclerosis (SSc) is a rare and severe disease characterized by generalized fibrosis, vasculopathy and deregulation of immune cells. No curative treatment is validated to date. Mesenchymal stromal cells (MSCs) are being evaluated in clinical trials as they are effective in several preclinical models of SSc. They exert their anti-fibrotic, pro-angiogenic and immunomodulatory properties through the secretion of mediators contained in extracellular vesicles (EVs). We investigate here the role of EVs released by MSCs in their systemic effect during SSc.In a murine model of hypochlorous acid-induced SSc, we demonstrate that EVs can stop the disease course and improve histological and molecular parameters in skin and lung samples. We identify miR-29a-3p expression in both MSCs and their EVs and prove that miR-29a-3p is responsible for a large part of their therapeutic effect, targeting Dnmt3a and Pdgfr-bb, two novel targets identified in SSc.We improve the therapeutic efficacy of EVs on lung abnormalities in SSc mice by stimulating MSCs with IFNγ before EVs production whereas increasing the amount of injected EVs did not have any benefit. IFNγ modulates the expression of known immunosuppressive factors in both MSCs and EVs, potentiating their therapeutic properties.While the role of MSCs from SSc patients (SSc MSCs) in the development of the disease is debated, we show that SSc MSCs inhibit in vitro T lymphocyte proliferation and improve the myofibroblastic phenotype obtained after fibroblast stimulation using TGFβ1 which display a similar profile as fibroblasts from SSc patients in the in vitro model used here.Overall, MSC-EVs are as effective as MSCs to stop the progression of murine SSc and EVs from IFNγ-preconditioned MSCs improve their beneficial effect in the lungs. SSc MSCs maintain their anti-fibrotic and immunosuppressive properties in vitro and one of our perspectives is to evaluate their therapeutic efficacy in vivo

Effet thérapeutique des vésicules extracellulaires isolées à partir de cellules stromales mésenchymateuses dans la sclérodermie systémique

Systemic sclerosis (SSc) is a rare and severe disease characterized by generalized fibrosis, vasculopathy and deregulation of immune cells. No curative treatment is validated to date. Mesenchymal stromal cells (MSCs) are being evaluated in clinical trials as they are effective in several preclinical models of SSc. They exert their anti-fibrotic, pro-angiogenic and immunomodulatory properties through the secretion of mediators contained in extracellular vesicles (EVs). We investigate here the role of EVs released by MSCs in their systemic effect during SSc.In a murine model of hypochlorous acid-induced SSc, we demonstrate that EVs can stop the disease course and improve histological and molecular parameters in skin and lung samples. We identify miR-29a-3p expression in both MSCs and their EVs and prove that miR-29a-3p is responsible for a large part of their therapeutic effect, targeting Dnmt3a and Pdgfr-bb, two novel targets identified in SSc.We improve the therapeutic efficacy of EVs on lung abnormalities in SSc mice by stimulating MSCs with IFNγ before EVs production whereas increasing the amount of injected EVs did not have any benefit. IFNγ modulates the expression of known immunosuppressive factors in both MSCs and EVs, potentiating their therapeutic properties.While the role of MSCs from SSc patients (SSc MSCs) in the development of the disease is debated, we show that SSc MSCs inhibit in vitro T lymphocyte proliferation and improve the myofibroblastic phenotype obtained after fibroblast stimulation using TGFβ1 which display a similar profile as fibroblasts from SSc patients in the in vitro model used here.Overall, MSC-EVs are as effective as MSCs to stop the progression of murine SSc and EVs from IFNγ-preconditioned MSCs improve their beneficial effect in the lungs. SSc MSCs maintain their anti-fibrotic and immunosuppressive properties in vitro and one of our perspectives is to evaluate their therapeutic efficacy in vivo.La sclérodermie systémique (ScS) est une maladie rare et sévère, caractérisée par une dérégulation des cellules immunitaires, une fibrose et une vasculopathie généralisées. Aucun traitement curatif n’est validé à ce jour. Les cellules stromales mésenchymateuses (CSMs), dont l’efficacité a été démontrée dans plusieurs modèles précliniques de ScS, sont actuellement évaluées dans des essais cliniques. Elles exercent leurs propriétés régénératrices au travers de la sécrétion de médiateurs libérés dans le milieu extracellulaire ou contenus dans des vésicules extracellulaires (EVs). Nous nous intéressons ici au rôle des EVs libérées par les CSMs dans leur effet systémique au cours de la ScS.Dans un modèle murin de ScS induit par des injections d’acide hypochloreux, nous démontrons que les EVs stoppent l’évolution de la maladie au niveau cutané et pulmonaire sur le plan clinique, histologique et moléculaire. Nous identifions miR-29a-3p au sein des CSMs et de leurs EVs et prouvons qu’il est garant de leur effet thérapeutique, en ciblant Dnmt3a et Pdgfr-bb, deux nouvelles cibles identifiées dans la ScS.Nous améliorons l’efficacité thérapeutique des EVs sur les anomalies pulmonaires en pré-activant les CSMs à l’aide d’IFNγ alors que l’augmentation de la quantité d’EVs injectées n’a aucun effet. L’IFNγ module l’expression de molécules immunosuppressives dans les CSMs et leurs EVs, potentialisant ainsi leurs propriétés thérapeutiques.Alors que le rôle des CSMs de patients (CSMs-ScS) dans le développement de la maladie est sujet à débat, nous montrons que les CSMs-ScS inhibent in vitro la prolifération des lymphocytes T de manière similaire à des CSMs saines. En outre, elles sont capables d’améliorer le phénotype myofibroblastique de fibroblastes stimulés par du TGFβ1 qui présentent un comportement similaire à celui de fibroblastes de patients dans le modèle in vitro mis en place.En conclusion, les EVs sont aussi efficaces que les CSMs pour traiter la ScS murine et le pré-conditionnement des CSMs par l’IFNγ améliore leur effet au niveau pulmonaire. Les CSMs de patients conservent leurs fonctions anti-fibrotique et immunosuppressive in vitro et une de nos perspectives est d’évaluer leur efficacité thérapeutique in vivo

Mesenchymal Stem Cell-Derived Extracellular Vesicles: Opportunities and Challenges for Clinical Translation

International audienceExtracellular vesicles (EVs), including exosomes and microvesicles, derived from mesenchymal stem/stromal cells (MSCs) exert similar effects as their parental cells, and are of interest for various therapeutic applications. EVs can act through uptake by the target cells followed by release of their cargo inside the cytoplasm, or through interaction of membrane-bound ligands with receptors expressed on target cells to stimulate downstream intracellular pathways. EV-based therapeutics may be directly used as substitutes of intact cells or after modification for targeted drug delivery. However, for the development of EV-based therapeutics, several production, isolation, and characterization requirements have to be met and the quality of the final product has to be tested before its clinical implementation. In this review, we discuss the challenges associated with the development of EV-based therapeutics and the regulatory specifications for their successful clinical translation

Mesenchymal Stem Cell Derived Extracellular Vesicles in Aging

International audienceAging is associated with high prevalence of chronic degenerative diseases that take a large part of the increasing burden of morbidities in a growing demographic of elderly people. Aging is a complex process that involves cell autonomous and cell non-autonomous mechanisms where senescence plays an important role. Senescence is characterized by the loss of proliferative potential, resistance to cell death by apoptosis and expression of a senescence-associated secretory phenotype (SASP). SASP includes pro-inflammatory cytokines and chemokines, tissue-damaging proteases, growth factors; all contributing to tissue microenvironment alteration and loss of tissue homeostasis. Emerging evidence suggests that the changes in the number and composition of extracellular vesicles (EVs) released by senescent cells contribute to the adverse effects of senescence in aging. In addition, age-related alterations in mesenchymal stem/stromal cells (MSCs) have been associated to dysregulated functions. The loss of functional stem cells necessary to maintain tissue homeostasis likely directly contributes to aging. In this review, we will focus on the characteristics and role of EVs isolated from senescent MSCs, the potential effect of MSC-derived EVs in aging and discuss their therapeutic potential to improve age-related diseases

Mesenchymal Stem Cells in Systemic Sclerosis: Allogenic or Autologous Approaches for Therapeutic Use?

International audienceSystemic sclerosis (SSc) is a rare autoimmune disease, which is potentially lethal. The physiopathology of the disease is still incompletely elucidated although the role of fibroblasts, endothelial cells (ECs), immune cells. and the environment (i.e., oxidative stress) has been demonstrated. This is an intractable disease with an urgent need to provide better therapeutic options to patients. Mesenchymal stem cells (MSCs) represent a promising therapeutic approach thanks to the number of trophic and pleiotropic properties they exert. Among these, MSCs display anti-fibrotic, angiogenic, and immunomodulatory capacities that might be of interest in the treatment of SSc by acting on different processes that are dysregulated in the disease. In the recent years, the therapeutic effectiveness of MSCs has been demonstrated in different preclinical animal models and is being investigated in phase I clinical trials. Both allogenic and autologous transplantation of MSCs isolated from bone marrow or adipose tissue is being evaluated. The rationale for using allogenic MSCs in SSc, as well as in other autoimmune diseases, is based on the possibility that autologous MSCs might be altered in these diseases. In SSc, reports from the literature are controversial. Nevertheless, the role of the oxidative environment and of the crosstalk with neighboring cells (fibroblasts and ECs) on the functional properties of MSCs has been reported. Here, we review the preclinical and clinical data reporting the interest of MSC-based treatment in SSc and question the use of autologous or allogeneic MSCs in perspective of clinical applications

Le TGFBI sécrété par les cellules stromales mésenchymateuses améliore l'arthrose et est détecté dans les vésicules extracellulaires

International audienceMesenchymal stem/stromal cells (MSCs) are of interest in the context of osteoarthritis (OA) therapy. We previously demonstrated that TGFβ-induced gene product-h3 (TGFBI/BIGH3) is downregulated in human MSCs (hMSCs) from patients with OA, suggesting a possible link with their impaired regenerative potential. In this study, we investigated TGFBI contribution to MSC-based therapy in OA models. First, we showed that co-culture with murine MSCs (mMSCs) partly restored the expression of anabolic markers and decreased expression of catabolic markers in OA-like chondrocytes only upon priming by TGFβ3. Moreover, TGFβ3-primed hMSCs not only modulated the expression of anabolic and catabolic markers, but also decreased inflammatory factors. Then, we found that upon TGFBI silencing, mMSCs partly lost their inductive effect on chondrocyte anabolic markers. Injection of hMSCs in which TGFBI was silenced did not protect mice from OA development. Finally, we showed that MSC chondroprotection was attributed to the presence of TGFBI mRNA and protein in extracellular vesicles. Our findings suggest that TGFBI is a chondroprotective factor released by MSCs and an anabolic regulator of cartilage homeostasis

Lung Fibrosis Is Improved by Extracellular Vesicles from IFNγ-Primed Mesenchymal Stromal Cells in Murine Systemic Sclerosis

International audienceBackground: Systemic sclerosis (SSc) is a severe autoimmune disease for which mesenchymal stromal cells (MSCs)-based therapy was reported to reduce SSc-related symptoms in pre-clinical studies. Recently, extracellular vesicles released by MSCs (MSC-EVs) were shown to mediate most of their therapeutic effect. Here, we aimed at improving their efficacy by increasing the MSC-EV dose or by IFNγ-priming of MSCs.Methods: small size (ssEVs) and large size EVs (lsEVs) were recovered from murine MSCs that were pre-activated using 1 or 20 ng/mL of IFNγ. In the HOCl-induced model of SSc, mice were treated with EVs at day 21 and sacrificed at day 42. Lung and skin samples were collected for histological and molecular analyses.Results: increasing the dose of MSC-EVs did not add benefit to the dose previously reported to be efficient in SSc. By contrast, IFNγ pre-activation improved MSC-EVs-based treatment, essentially in the lungs. Low doses of IFNγ decreased the expression of fibrotic markers, while high doses improved remodeling and anti-inflammatory markers. IFNγ pre-activation upregulated iNos, IL1ra and Il6 in MSCs and ssEVs and the PGE2 protein in lsEVs.Conclusion: IFNγ-pre-activation improved the therapeutic effect of MSC-EVs preferentially in the lungs of SSc mice by modulating anti-inflammatory and anti-fibrotic markers

Extracellular Vesicles Are More Potent Than Adipose Mesenchymal Stromal Cells to Exert an Anti-Fibrotic Effect in an In Vitro Model of Systemic Sclerosis

International audienceSystemic sclerosis (SSc) is a complex disorder resulting from dysregulated interactions between the three main pathophysiological axes: fibrosis, immune dysfunction, and vasculopathy, with no specific treatment available to date. Adipose tissue-derived mesenchymal stromal cells (ASCs) and their extracellular vesicles (EVs) have proved efficacy in pre-clinical murine models of SSc. However, their precise action mechanism is still not fully understood. Because of the lack of availability of fibroblasts isolated from SSc patients (SSc-Fb), our aim was to determine whether a TGFβ1-induced model of human myofibroblasts (Tβ-Fb) could reproduce the characteristics of SSc-Fb and be used to evaluate the anti-fibrotic function of ASCs and their EVs. We found out that Tβ-Fb displayed the main morphological and molecular features of SSc-Fb, including the enlarged hypertrophic morphology and expression of several markers associated with the myofibroblastic phenotype. Using this model, we showed that ASCs were able to regulate the expression of most myofibroblastic markers on Tβ-Fb and SSc-Fb, but only when pre-stimulated with TGFβ1. Of interest, ASC-derived EVs were more effective than parental cells for improving the myofibroblastic phenotype. In conclusion, we provided evidence that Tβ-Fb are a relevant model to mimic the main characteristics of SSc fibroblasts and investigate the mechanism of action of ASCs. We further reported that ASC-EVs are more effective than parental cells suggesting that the TGFβ1-induced pro-fibrotic environment may alter the function of ASCs