Stem Cell Reviews and Reports

, Volume 12, Issue 4, pp 448–453

Transplantation of Human Adipose Mesenchymal Stem Cells in Non-Immunosuppressed GRMD Dogs is a Safe Procedure

  • M. V. Pelatti
  • J. P. A. Gomes
  • N. M. S. Vieira
  • E. Cangussu
  • V. Landini
  • T. Andrade
  • M. Sartori
  • L. Petrus
  • Mayana Zatz
Article

Abstract

The possibility to treat Duchenne muscular dystrophy (DMD), a lethal X-linked disorder, through cell therapy with mesenchymal stromal cells (MSCs) has been widely investigated in different animal models. However, some crucial questions need to be addressed before starting human therapeutic trials, particularly regarding its use for genetic disorders. How safe is the procedure? Are there any side effects following mesenchymal stem cell transplantation? To address these questions for DMD the best model is the golden retriever muscular dystrophy dog (GRMD), which is the closest model to the human condition displaying a much longer lifespan than other models. Here we report the follow-up of 5 GRMD dogs, which were repeatedly transplanted with human adipose-derived mesenchymal stromal cells (hASC), derived from different donors. Xenogeneic cell transplantation, which was done without immunosuppression, was well tolerated in all animals with no apparent long-term adverse effect. In the present study, we show that repeated heterologous stem-cell injection is a safe procedure, which is fundamental before starting human clinical trials.

Keywords

Human multipotent mesenchymal stromal cells Xenotransplantation Muscular dystrophy Cell therapy GRMD dogs 

Supplementary material

12015_2016_9659_MOESM1_ESM.pdf (516 kb)
Supplementary figure 1Smart Ultrasound evaluation 3 months after treatment. (PDF 516 kb)
12015_2016_9659_MOESM2_ESM.pdf (554 kb)
Supplementary figure 2Tutu Ultrasound evaluation 3 months after treatment. (PDF 554 kb)
12015_2016_9659_MOESM3_ESM.docx (1 mb)
Supplementary figure 3Smart x-ray evaluation 3 months after treatment. (DOCX 1044 kb)
12015_2016_9659_MOESM4_ESM.docx (1 mb)
Supplementary figure 4Tutu x-ray evaluation 3 months after treatment. (DOCX 1038 kb)
12015_2016_9659_MOESM5_ESM.pdf (505 kb)
Supplementary figure 5Yuan Ultrasound evaluation 61/2 years after treatment. (PDF 504 kb)
12015_2016_9659_MOESM6_ESM.docx (867 kb)
Supplementary figure 6Yuan x-ray evaluation 61/2 years after treatment. (DOCX 866 kb)
12015_2016_9659_MOESM7_ESM.pdf (527 kb)
Supplementary figure 7Rum Ultrasound evaluation 1 year after treatment. (PDF 526 kb)
12015_2016_9659_MOESM8_ESM.docx (1.2 mb)
Supplementary figure 8Rum x-ray evaluation. (DOCX 1220 kb)
12015_2016_9659_MOESM9_ESM.pdf (441 kb)
Supplementary table 1Cardiac evaluations of Sushi (normal dog), Smart and Tutu before and after treatment. Cardiac data include electrocardiography, mean arterial pressure and echocardiography. (PDF 441 kb)
12015_2016_9659_MOESM10_ESM.pdf (291 kb)
Supplementary table 2Complete blood test, including blood count and biochemical blood test of Sushi (normal dog), Smart and Tutu. Blood was collected before and 48 h after each stem cell injection. Presented data show minimum and maximum values observed in all 18 blood tests. (PDF 290 kb)
12015_2016_9659_MOESM11_ESM.pdf (201 kb)
Supplementary table 3Last complete blood test of Sushi (normal dog), Smart, Tutu, Yuan and Rum. Evaluation was performed 6 month after Tutu and Smart treatment; 10 months after Rum treatment; and 61/2year after Yuan treatment. (PDF 200 kb)

References

  1. 1.
    Wicklund, M. P. (2013). The muscular dystrophies. continuum (Minneapolis Minnesota), 19, 1535–1570.Google Scholar
  2. 2.
    Emery, A. E. H. (2002). The muscular dystrophies. Lancet, 359, 687–695.CrossRefPubMedGoogle Scholar
  3. 3.
    Vainzof, M., et al. (2008). Animal models for genetic neuromuscular diseases. Journal of Molecular Neuroscience, 34, 241–248.CrossRefPubMedGoogle Scholar
  4. 4.
    Allamand, V., & Campbell, K. P. (2000). Animal models for muscular dystrophy: valuable tools for the development of therapies. Human Molecular Genetics, 9, 2459–2467.CrossRefPubMedGoogle Scholar
  5. 5.
    Sharp, N. J., et al. (1992). An error in dystrophin mRNA processing in golden retriever muscular dystrophy, an animal homologue of Duchenne muscular dystrophy. Genomics, 13, 115–121.CrossRefPubMedGoogle Scholar
  6. 6.
    Zatz, M., et al. (2014). Milder course in Duchenne patients with nonsense mutations and no muscle dystrophin. Neuromuscular Disorders, 24, 986–989.CrossRefPubMedGoogle Scholar
  7. 7.
    Zucconi, E., et al. (2010). Ringo: discordance between the molecular and clinical manifestation in a golden retriever muscular dystrophy dog. Neuromuscular Disorders, 20, 64–70.CrossRefPubMedGoogle Scholar
  8. 8.
    Zatz, M., et al. (2015). A normal life without muscle dystrophin. Neuromuscular Disorders. doi:10.1016/j.nmd.2015.02.007.Google Scholar
  9. 9.
    Vieira, N. M., et al. (2015). Muscular dystrophy in a family of labrador retrievers with no muscle dystrophin and a mild phenotype. Neuromuscular Disorders. doi:10.1016/j.nmd.2015.02.012.Google Scholar
  10. 10.
    Quattrocelli, M., Cassano, M., Crippa, S., Perini, I., & Sampaolesi, M. (2010). Cell therapy strategies and improvements for muscular dystrophy. Cell Death and Differentiation, 17, 1222–1229.CrossRefPubMedGoogle Scholar
  11. 11.
    Sampaolesi, M., et al. (2006). Mesoangioblast stem cells ameliorate muscle function in dystrophic dogs. Nature, 444, 574–579.CrossRefPubMedGoogle Scholar
  12. 12.
    Nitahara-Kasahara, Y., et al. (2012). Long-term engraftment of multipotent mesenchymal stromal cells that differentiate to form myogenic cells in dogs with Duchenne muscular dystrophy. Molecular Therapy, 20, 168–177.CrossRefPubMedGoogle Scholar
  13. 13.
    Rouger, K., et al. (2011). Systemic delivery of allogenic muscle stem cells induces long-term muscle repair and clinical efficacy in duchenne muscular dystrophy dogs. The American Journal of Pathology, 179, 2501–2518.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Cerletti, M., et al. (2008). Highly efficient, functional engraftment of skeletal muscle stem cells in dystrophic muscles. Cell, 134, 37–47.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Valadares, M. C., et al. (2014). Human adipose tissue derived pericytes increase life span in Utrn (tm1Ked) Dmd (mdx) /J mice. Stem Cell Reviews, 10, 830–840.CrossRefPubMedGoogle Scholar
  16. 16.
    Vieira, N. M., et al. (2012). Human adipose-derived mesenchymal stromal cells injected systemically into GRMD dogs without immunosuppression are able to reach the host muscle and express human dystrophin. Cell Transplantation, 21, 1407–1417.CrossRefPubMedGoogle Scholar
  17. 17.
    Honeyman, K., Carville, K. S., Howell, J. M., Fletcher, S., & Wilton, S. D. (1999). Development of a snapback method of single-strand conformation polymorphism analysis for genotyping Golden Retrievers for the X-linked muscular dystrophy allele. American Journal of Veterinary Research, 60, 734–737.PubMedGoogle Scholar
  18. 18.
    Vieira, N. M., et al. (2008). SJL dystrophic mice express a significant amount of human muscle proteins following systemic delivery of human adipose-derived stromal cells without immunosuppression. Stem Cells, 26, 2391–2398.CrossRefPubMedGoogle Scholar
  19. 19.
    Zuk, P. A., et al. (2002). Human adipose tissue is a source of multipotent stem cells. Molecular Biology of the Cell, 13, 4279–4295.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Caplan, A. I. (1991). Mesenchymal stem cells. Journal of Orthopaedic Research, 9, 641–650.CrossRefPubMedGoogle Scholar
  21. 21.
    Dominici, M., et al. (2006). Minimal criteria for defining multipotent mesenchymal stromal cells. The international society for cellular therapy position statement. Cytotherapy, 8, 315–317.CrossRefPubMedGoogle Scholar
  22. 22.
    Caplan, A. I., & Dennis, J. E. (2006). Mesenchymal stem cells as trophic mediators. Journal of Cellular Biochemistry, 98, 1076–1084.CrossRefPubMedGoogle Scholar
  23. 23.
    Singer, N. G., & Caplan, A. I. (2011). Mesenchymal stem cells: mechanisms of inflammation. Annual Review of Pathology, 6, 457–478.CrossRefPubMedGoogle Scholar
  24. 24.
    Caplan, A. I., & Sorrell, J. M. (2015). The MSC curtain that stops the immune system. Immunology Letters. doi:10.1016/j.imlet.2015.06.005.PubMedGoogle Scholar
  25. 25.
    Ichim, T. E., et al. (2010). Mesenchymal stem cells as anti-inflammatories: implications for treatment of Duchenne muscular dystrophy. Cellular Immunology, 260, 75–82.CrossRefPubMedGoogle Scholar
  26. 26.
    Le Blanc, K. (2006). Mesenchymal stromal cells: tissue repair and immune modulation. Cytotherapy, 8, 559–561.CrossRefPubMedGoogle Scholar
  27. 27.
    Le Blanc, K., Tammik, L., Sundberg, B., Haynesworth, S. E., & Ringdén, O. (2003). Mesenchymal stem cells inhibit and stimulate mixed lymphocyte cultures and mitogenic responses independently of the major histocompatibility complex. Scandinavian Journal of Immunology, 57, 11–20.CrossRefPubMedGoogle Scholar
  28. 28.
    English, K. (2013). Mechanisms of mesenchymal stromal cell immunomodulation. Immunology and Cell Biology, 91, 19–26.CrossRefPubMedGoogle Scholar
  29. 29.
    N. M. Vieira et al. (2015) Jagged 1 Rescues the Duchenne Muscular Dystrophy Phenotype. Cell, 1–10.Google Scholar
  30. 30.
    B. Gharaibeh, M. Lavasani, J. H. Cummins, J. Huard. (2011). Terminal differentiation is not a major determinant for the success of stem cell therapy - cross-talk between muscle-derived stem cells and host cells. Stem Cell Research & Therapy.Google Scholar
  31. 31.
    Pinheiro, C. H. D. J., et al. (2012). Local injections of adipose-derived mesenchymal stem cells modulate inflammation and increase angiogenesis ameliorating the dystrophic phenotype in dystrophin-deficient skeletal muscle. Stem Cell Reviews, 8, 363–374.CrossRefPubMedGoogle Scholar
  32. 32.
    Murphy, M. B., Moncivais, K., & Caplan, A. I. (2013). Mesenchymal stem cells: environmentally responsive therapeutics for regenerative medicine. Experimental and Molecular Medicine, 45, e54.CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Lee, S.-R., et al. (2015). Long-term survival and differentiation of human neural stem cells in nonhuman primate brain with no immunosuppression. Cell Transplantation, 24, 191–201.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • M. V. Pelatti
    • 1
  • J. P. A. Gomes
    • 1
  • N. M. S. Vieira
    • 1
  • E. Cangussu
    • 1
  • V. Landini
    • 1
  • T. Andrade
    • 1
  • M. Sartori
    • 1
  • L. Petrus
    • 1
  • Mayana Zatz
    • 1
  1. 1.Human Genome and Stem-cell Research Center, Institute of BiosciencesUniversity of São PauloSão PauloBrasil

Personalised recommendations