Elsevier

Transplant Immunology

Volume 55, August 2019, 101205
Transplant Immunology

Adipose tissue-derived mesenchymal stem cells ameliorate bone marrow aplasia related with graft-versus-host disease in experimental murine models

https://doi.org/10.1016/j.trim.2019.03.004Get rights and content

Highlights

  • Adipose tissue-derived mesenchymal stem cells (AD-MSCs) control GVHD in mouse models.

  • In particular, AD-MSCs protect against bone marrow aplasia related with GVHD in mice.

  • AD-MSCs may be useful to ameliorate bone marrow aplasia related with GVHD in humans.

Abstract

Graft-versus-host disease (GVHD) constitutes the most frequent complications after the allogeneic hematopoietic stem cell transplantation for a variety of hematological malignancies. In the present study, we explored the prophylactic potential of adipose tissue-derived mesenchymal stem cells (AD-MSCs) in controlling GVHD in murine models with a special focus on bone marrow aplasia related with acute GVHD. The CB6F1 mice were induced GVHD by the injection intravenously of C57BL/6 (B6-Ly-5.1) splenocytes without conditioning irradiation or chemotherapy. AD-MSCs from C3H mice were injected intravenously via tail veins. GVHD was assessed using flowcytometry analysis of peripheral blood cells and histopathologic analysis of target organs. Histopathological analyses revealed that AD-MSCs markedly suppressed the infiltration of lymphocytes into liver as well as the aplasia in bone marrow. This study is the first to clarify the effectiveness of AD-MSCs against bone marrow aplasia in GVHD, supporting a rationale of AD-MSCs for ameliorating bone marrow suppression and infectivity after allo-HSCT in human clinics.

Introduction

Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is a curative therapy with proven efficacy in the management of hematologic malignance. In some cases, however, it is complicated by acute and chronic graft-versus-host disease (GVHD) [[1], [2], [3]]. Outside of primary disease relapse following allo-HSCT, arguably the most obstacle to success following this therapy is the onset of GVHD [4]. Although corticosteroids are used as a first-line therapy [5], only 60–70% of patients with acute GVHD respond to standard corticosteroid therapy, and those who failed initial therapy showed only a 10–30% chance of long-term survival [4,6,7]. A wide variety of second-line treatments for acute GVHD are available including tacrolimus, mycophenolate mofetil (MMF), sirolimus, anti-thymocyte globulin (ATG), monoclonal antibodies against IL-2 receptor, TNFα, CD52, CD147, and CD3 [[8], [9], [10], [11], [12]]. However, the prognosis of patients under these treatments remains dismal because of higher risk of infectious complications, immunosuppression-mediated toxicity, and incomplete remission of GVHD [[8], [9], [10], [11], [12]]. Therefore, better strategies for the prophylaxis and treatment of GVHD are critical to improve the outcome of HSCT.

It is well known that the principal target organs of acute GVHD are the skin, liver and gastrointestinal tract. Moreover, GVHD is often accompanied by cytopenia and bone marrow suppression [13]. In a murine model of GVHD, Shono et al. reported the destruction of bone marrow hematopoietic niches, especially osteoblasts by donor T cells, resulting in bone marrow suppression [13], an example of bone marrow GVHD. Because bone marrow GVHD may cause life-threatening infection after the allo-HSCT [14], bone marrow destruction caused by acute GVHD should be prevented in clinics.

Mesenchymal stromal cells (MSCs) are a form of multipotent adult stem cells characterized by their plastic-adherent growth and expression of specific surface antigen [15]. MSCs possess the capacity to suppress immunological responses, support hematopoiesis and repair tissues [[16], [17], [18], [19]]. Based on a variety of immunoregulatory properties, MSCs (bone marrow derived MSCs) have been studied as a promising platform for cell-based therapies to prevent or treat GVHD [[20], [21], [22]]. MSCs are present in various tissues including bone marrow, peripheral bloods, umbilical cord blood and adipose tissue. It has been highlighted that adipose tissue derived MSCs (AD-MSCs) might serve as promising therapeutic tools for GVHD with a line of unique advantage [23]. In particular, large amount of adipose tissues can be obtained in a minimally invasive manner through lipectomy or liposuction. Furthermore, previous in vitro studies showed that AD-MSCs represent equipotent or higher immunoregulatory capacity compared to bone marrow derived MSCs [24].

Although many studies reported that MSCs ameliorated GVHD, to our knowledge, there have been no reports focusing on the impact of MSCs on bone marrow aplasia related with GVHD. There are a large number of murine acute GVHD models, and most of them use lethally irradiation regimen prior to transplantation. On the other hand, in a MHC-mismatched murine model, parent-to-F1 transplants have been studied using no irradiation conditioning [25,26]. As pretreating chemotherapy and irradiation are responsible for bone marrow destruction [14], it has been difficult to analyze bone marrow per se in preexisting GVHD murine models. The model without irradiation is advantageous for assessing the impact of GVHD itself on bone marrow. In this context, we employed murine GVHD model of MHC homozygous-donor to heterozygous-recipient without pretreating chemotherapy or irradiation.

Section snippets

Animals

C57BL/6 J(H-2b), (BALB/c × C57BL/6) F1 (CB6F1) (H-2dXb) and C3H/He(H-2k) female mice of 6–8 weeks of age were purchased from Japan SLC, Inc. (Shizuoka, Japan). This study was conducted in compliance with the Animal Welfare Act, the implementing Animal Welfare Regulations and in accordance with the principles of the Guide for the Care and Use of Laboratory Animals. Study protocol was reviewed and approved by the Animal Experiment Ethics Committee of the University of the Ryukyus (A2016029).

Isolation, culture, and in vitro differentiation of AD-MSCs

To

Characterization of isolated AD-MSCs

AD-MSCs were isolated from adipose tissues obtained from the inguinal region of C3H/He mice. The cultured AD-MSCs exhibited a fibroblast-like, spindle-shaped morphology (Fig. 1A). Flow cytometry analysis was used to assess whether the AD-MSCs showed typical profile of MSC surface marker. The AD-MSCs highly expressed a variety of markers for authentic mesenchymal stem cells including Scasingle bondI, CD29, CD90 and CD105, while they were negative for hematopoietic and endothelial markers such as CD31 and

Discussion

The present study demonstrated that bone marrow destruction induced by acute GVHD was potently protected by the administration of AD-MSCs, where inflammation or bone marrow destruction caused by preconditioning (i.e. irradiation or chemotherapy) was absent. We used murine allo-HSCT model of MHC homo-donor to hetero-recipient without preconditioning therapy, because this model is likely to correspond to transfusion-associated GVHD models in humans rather than allo-HSCT. In most cases,

Conflict of interest

The authors report no potential conflict of interest.

Authors' contributions

Y. N. and A. M. performed the research, analyzed the data and wrote the manuscript; Y. M. performed the research; N. T. analyzed the data, S. O., S. N., H. N., M. M., T. F., K. K. and S. M. wrote the manuscript. H. K. and H. M. supervised the research, analyzed the data and wrote the manuscript.

Funding

This work was supported in part by grants-in-Aid from Japan Society for the Promotion of Science (JSPS; KAKENHI Grant Numbers 15K19520 and 24591338), the Council for Science, Technology and Innovation (CSTI), Cross-ministerial Strategic Innovation Promotion Program (SIP) ‘Technologies for creating next-generation agriculture, forestry and fisheries’, the Lotte Foundation, the Japan Foundation for Applied Enzymology, New Energy and Industrial Technology Development Organization (NEDO), the

Acknowledgments

We would like to express my gratitude to T. Teshima and D. Hashimoto, Department of Hematology, Hokkaido University Graduate School of Medicine, for technical support. We are grateful to S. Kitamura, S Uchibori, S Tomori, T Hanashiro, N Shimabukuro and I Tedokon for reviewing the manuscript. We thank M. Hirata, H. Kaneshiro, I. Asato and C. Noguchi for secretarial assistance.

References (44)

  • K. Le Blanc et al.

    Mesenchymal stem cells for treatment of steroid-resistant, severe, acute graft-versus-host disease: a phase II study

    Lancet

    (2008)
  • P. Kebriaei et al.

    Adult human mesenchymal stem cells added to corticosteroid therapy for the treatment of acute graft-versus-host disease

    Biol. Blood Marrow Transplant.

    (2009)
  • J. Kurtzberg et al.

    Allogeneic human mesenchymal stem cell therapy (remestemcel-L, Prochymal) as a rescue agent for severe refractory acute graft-versus-host disease in pediatric patients

    Biol. Blood Marrow Transplant.

    (2014)
  • K.R. Cooke et al.

    An experimental model of idiopathic pneumonia syndrome after bone marrow transplantation: I. The roles of minor H antigens and endotoxin

    Blood

    (1996)
  • I. Kopolovic et al.

    A systematic review of transfusion-associated graft-versus-host disease

    Blood

    (2015)
  • B. Fang et al.

    Favorable response to human adipose tissue-derived mesenchymal stem cells in steroid-refractory acute graft-versus-host disease

    Transplant. Proc.

    (2007)
  • B. Fang et al.

    Using human adipose tissue-derived mesenchymal stem cells as salvage therapy for hepatic graft-versus-host disease resembling acute hepatitis

    Transplant. Proc.

    (2007)
  • M. Jurado et al.

    Adipose tissue-derived mesenchymal stromal cells as part of therapy for chronic graft-versus-host disease: a phase I/II study

    Cytotherapy

    (2017)
  • M. Rizk et al.

    Heterogeneity in studies of mesenchymal stromal cells to treat or prevent graft-versus-host disease: a scoping review of the evidence

    Biol. Blood Marrow Transplant.

    (2016)
  • M. Rodbell

    Metabolism of isolated fat cells. I. Effects of hormones on glucose METABOLISM and lipolysis

    J. Biol. Chem.

    (1964)
  • J.H. Sung et al.

    Isolation and characterization of mouse mesenchymal stem cells

    Transplant. Proc.

    (2008)
  • Y. Muguruma et al.

    Reconstitution of the functional human hematopoietic microenvironment derived from human mesenchymal stem cells in the murine bone marrow compartment

    Blood

    (2006)
  • Cited by (7)

    • Umbilical cord-derived mesenchymal stem cells promote myeloid-derived suppressor cell enrichment by secreting CXCL1 to prevent graft-versus-host disease after hematopoietic stem cell transplantation

      2021, Cytotherapy
      Citation Excerpt :

      Dermis-derived mesenchymal stem cells (DMSCs) may control refractory GVHD by preventing peripheral blood mononuclear cells from entering S phase, suppressing the activation of CD3+ T cells and increasing the proportion of regulatory T cells (Tregs) [10]. Adipose tissue-derived mesenchymal stem cells (ADSCs) suppress the infiltration of lymphocytes into the liver and also suppress bone marrow (BM) aplasia and show potential for the treatment of GVHD [11]. The human umbilical cord is an alternative source of MSCs with several advantages, including easy access as a discarded tissue, low risk of viral infection, rich source of materials and lack of ethical concerns [12].

    • IFN-γ gene loaded human umbilical mesenchymal stromal cells targeting therapy for Graft-versus-host disease

      2021, International Journal of Pharmaceutics
      Citation Excerpt :

      Shono et al. reported the destruction of bone marrow hematopoietic niches in a murine model of GVHD, especially osteoblasts by donor T cells, resulting in bone marrow suppression. ( Nishi et al., 2019; Shono et al., 2010) Therefore, we analyzed the potential effect of HUMSCs + Ad-h/mIFN-γ on cell counts in peripheral blood of recipient mice. The blood sample was taken from the orbital vein weekly.

    • Phytocannabinoids promote viability and functional adipogenesis of bone marrow-derived mesenchymal stem cells through different molecular targets

      2020, Biochemical Pharmacology
      Citation Excerpt :

      The results presented in this paper suggest that select non-euphoric phytocannabinoids, i.e. CBDA, CBG or CBD, either individually or in combination, modulate multiple relevant receptors, including CB2 (possibly indirectly, since these compounds do not possess agonist activity at CB2 at the concentrations used in this study, so further investigations are required) and PPARγ, whose simultaneous activation appear to demonstrate additivity, promoting the targeted enhancement of viable endogenous BM-MSCs and their adipogenic potential, respectively. This may bypass the problems associated with exogenous MSC delivery [89] by actively directing MSC lineage fate to transition into committed adipocyte progenitors and the subsequent enhancement of adipogenic induction capacity, selecting for specific functional mature adipocyte subtypes, e.g. white, beige or brown adipocytes. The promiscuous pharmacology of phytocannabinoids makes them viable candidates for new medicines for the treatment of metabolic syndromes through the simultaneous resolution of collective complications due to impaired development, maintenance, activity and function of the adipose tissue.

    View all citing articles on Scopus
    1

    These authors contributed equally to this work.

    View full text