Early transplantation of human immature dental pulp stem cells from baby teeth to golden retriever muscular dystrophy (GRMD) dogs: Local or systemic?

Journal of Translational Medicine - 2008
Irina Kerkis1, Carlos Eduardo Ambrósio2, Alexandre Kerkis3, Daniel Lahan Martins2, Eder Zucconi4, Silvia Moulin Ribeiro Fonseca1, Richard Haltí Cabral2, C. M. C. Maranduba1, Thaís Peixoto Gaiad2, Adriana Caroprezo Morini2, N.M. Vieira4, Marina Pandolphi Brólio2, Osvaldo A. Sant’Anna1, María Angélica Miglino2, Mayana Zatz4
1Laboratório de Genética e Imunoquímica, Instituto Butantan, São Paulo, SP, Brasil
2Departamento de Cirurgia da Faculdade de Medicina Veterinária da Universidade de São Paulo, SP, Brasil
3Genética Aplicada, Atividades Veterinárias LTD, São Paulo, SP, Brazil
4Centro de Estudos do Genoma Humano, Departamento de Genética e Biologia Evolutiva, Universidade de São Paulo, SP, Brasil

Tóm tắt

AbstractBackgroundThe golden retriever muscular dystrophy (GRMD) dogs represent the best available animal model for therapeutic trials aiming at the future treatment of human Duchenne muscular dystrophy (DMD). We have obtained a rare litter of six GRMD dogs (3 males and 3 females) born from an affected male and a carrier female which were submitted to a therapeutic trial with adult human stem cells to investigate their capacity to engraft into dogs muscles by local as compared to systemic injection without any immunosuppression.MethodsHuman Immature Dental Pulp Stem Cells (hIDPSC) were transplanted into 4 littermate dogs aged 28 to 40 days by either arterial or muscular injections. Two non-injected dogs were kept as controls. Clinical translation effects were analyzed since immune reactions by blood exams and physical scores capacity of each dog. Samples from biopsies were checked by immunohistochemistry (dystrophin markers) and FISH for human probes.Results and DiscussionWe analyzed the cells' ability in respect to migrate, engraftment, and myogenic potential, and the expression of human dystrophin in affected muscles. Additionally, the efficiency of single and consecutive early transplantation was compared. Chimeric muscle fibers were detected by immunofluorescence and fluorescentin situhybridisation (FISH) using human antibodies and X and Y DNA probes. No signs of immune rejection were observed and these results suggested that hIDPSC cell transplantation may be done without immunosuppression. We showed that hIDPSC presented significant engraftment in GRMD dog muscles, although human dystrophin expression was modest and limited to several muscle fibers. Better clinical condition was also observed in the dog, which received monthly arterial injections and is still clinically stable at 25 months of age.ConclusionOur data suggested that systemic multiple deliveries seemed more effective than local injections. These findings open important avenues for further researches.

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Tài liệu tham khảo

Fadic R, Mezzano V, Alvarez K, Cabrera D, Holmgren J, Brandan E: Increase in decorin and biglycan in Duchenne Muscular Dystrophy: role of fibroblasts as cell source of these proteoglycans in the disease. J Cell Mol Med. 2006, 10: 758-69. 10.1111/j.1582-4934.2006.tb00435.x.

Mauro A: Satellite cell of skeletal muscle fibers. J Biophys Biochem Cytol. 1961, 9: 493-95.

Zammi P, Beauchamp J: The skeletal muscle stem cells or son of stem cells?. Differentiation. 2001, 68: 193-204. 10.1046/j.1432-0436.2001.680407.x.

Gussoni E, Soneoka Y, Strickland CD, Buzney EA, Khan MK, Flint AF, Kunkel LM, Mulligan RC: Dystrophin expression in the mdx mouse restored by stem cell transplantation. Nature. 1999, 401: 390-94.

Stedman HH, Sweeney HL, Shrager JB, Maguire HC, Panettieri RA, Petrof B, Narusawa M, Leferovich JM, Sladky JT, Kelly AM: The mdx mouse diaphragm reproduces the degenerative changes of Duchenne muscular dystrophy. Nature. 1991, 352: 536-39. 10.1038/352536a0.

Bach AD, Beier JP, Stern-Staeter J, Horch RE: Skeletal muscle tissue engineering. J Cell Mol Med. 2004, 8: 413-22. 10.1111/j.1582-4934.2004.tb00466.x.

Camirand G, Rousseau J, Ducharme MÈ, Rothstein DM, Tremblay JP: Novel Duchenne muscular dystrophy treatment through myoblast transplantation tolerance with anti-CD45RB, anti-CD154 and mixed chimerism. Am J Transplant. 2004, 4: 1255-265. 10.1111/j.1600-6143.2004.00501.x.

Sharp NJ, Kornegay JN, Van Camp SD, Herbstreith MH, Secore SL, Kettle S, Hung WY, Constantinou CD, Dykstra MJ, Roses AD: An error in dystrophin mRNA processing causes golden retriever muscular dystrophy, an animal homologue of Duchenne muscular dystrophy. Genomics. 1992, 13: 115-21. 10.1016/0888-7543(92)90210-J.

Valentine BA, Winand NJ, Pradhan D, Moise NS, de Lahunta A, Kornegay JN, Cooper BJ: Canine X-linked muscular dystrophy as an animal model of Duchenne muscular dystrophy: a review. Am J Med Genet. 1992, 42: 352-56. 10.1002/ajmg.1320420320.

Dell'Agnola C, Wang Z, Storb R, Tapscott SJ, Kuhr CS, Hauschka SD, Lee RS, Sale GE, Zellmer E, Gisburne S, Bogan J, Kornegay JN, Cooper BJ, Gooley TA, Little MT: Hematopoietic stem cell transplantation does not restore dystrophin expression in Duchenne muscular dystrophy dogs. Blood. 2004, 104: 4311-318. 10.1182/blood-2004-06-2247.

Sampaolesi M, Blot S, D'Antona G, Granger N, Tonlorenzi R, Innocenzi A, Mognol P, Thibaud JL, Galvez BG, Barthelemy I, Perani L, Mantero S, Guttinger M, Pansarasa O, Rinaldi C, Cusella De Angelis MG, Torrente Y, Bordignon C, Bottinelli R, Cossu G: Mesoangioblast stem cells ameliorate muscle function in dystrophic dogs. Nature. 2006, 444: 574-79. 10.1038/nature05282.

Grounds MD, Davies KE: The allure of stem cell therapy for muscular dystrophy. Neuromuscul Disord. 2007, 17: 206-8. 10.1016/j.nmd.2007.01.007.

Kerkis I, Kerkis A, Dozortsev D, Stukart-Parsons GC, Gomes Massironi SM, Pereira LV, Caplan AI, Cerruti HF: Isolation and characterization of a population of immature dental pulp stem cells expressing OCT-4 and other embryonic stem cells markers. Cells Tissues Organs. 2000, 184: 105-16. 10.1159/000099617.

de Mendonça Costa A, Bueno DF, Martins MT, Kerkis I, Kerkis A, Fanganiello RD, Cerruti H, Alonso N, Passos-Bueno MR: Reconstruction of large cranial defects in nonimmunosuppressed experimental design with human dental pulp stem cells. J Craniofacial Surg. 2008, 19: 204-10.

Pierdomenico L, Bonsi L, Calvitti M, Rondelli D, Arpinati M, Chirumbolo G, Becchetti E, Marchionni C, Alviano F, Fossati V, Staffolani N, Franchina M, Grossi A, Bagnara GP: Multipotent mesenchymal stem cells with immunosuppressive activity can be easily isolated from dental pulp. Transplantation. 2005, 80: 836-42. 10.1097/01.tp.0000173794.72151.88.

Di Nicola M, Carlo-Stella C, Magni M, Milanesi M, Longoni PD, Matteucci P, Grisanti S, Gianni AM: Human bone marrow stromal cells supress T-Lymphocyte proliferation induced by cellular or nonspecific mitogenic stimuli. Blood. 2002, 99: 3838-843. 10.1182/blood.V99.10.3838.

Gandia C, Armiñan A, García-Verdugo JM, Lledó E, Ruiz A, Miñana MD, Sanchez-Torrijos J, Payá R, Mirabet V, Carbonell-Uberos F, Llop M, Montero JA, Sepúlveda P: Human dental pulp stem cells improve left ventricular function, induce angiogenesis and reduce infarct size in rats with acute myocardial infarction. Stem Cells. 2008, 26: 638-45. 10.1634/stemcells.2007-0484.

Honeyman K, Carville KS, Howell JM, Fletcher S, Wilton SD: Development of a snapback method of single-strand conformation polymorphism analysis for genotyping Golden Retrievers for the X-linked muscular dystrophy allele. Am J Vet Res. 1999, 60: 734-37.

Vainzof M, Passos-Bueno MR, Zatz M: Immunological methods for the analysis of protein expression in neurmuscular diseases. 2003, Potter, NT: Methods in Molecular Medicine-Neurogenetics: Methods and Protocols. Humana Press Inc., Totowa, NJ, 355-78.

Thanh LT, Neguyen TM, Helliwell TR, Morris GE: Characterization of revertant muscle fibers in Duchenne Muscular Dystrophy using exon-specific monoclonal antibodies against dystrophin. Am J Hum Genet. 1995, 56: 725-31.

Thibaud JL, Monnet A, Bertoldi D, Barthélély I, Blot S, Carlier PG: Characterization of dystrophic muscle in golden retriever muscular dystrophy dogs by nuclear magnetic resonance imaging. Neuromuscul Disord. 2007, 17: 569-75. 10.1016/j.nmd.2007.03.004.

Zhang W, Walboomers XF, Shi S, Fan M, Jansen JA: Multilineage differentiation potential of stem cells derived from dental pulp after cryopreservation. Tissue Eng. 2006, 12: 2813-823. 10.1089/ten.2006.12.2813.

DiDio LJA: Tratado de anatomia aplicada. Póllus Editorial, São Paulo. 1998, 103-109.

Zatz M, Rapaport D, Vainzof M, Rocha JML, Passos-Bueno MR, Rabbi-Bortolini E, Pavanello RCM, Peres CA: Serum creatine-kinase (CK) and pyruvate-kinase (PK) activity as a function of clinical evolution in Duchenne (DMD) and Becker (BMD) muscular dystrophies. J Neurol Sci. 1991, 102: 190-196. 10.1016/0022-510X(91)90068-I.

Tse WT, Pendleton JD, Beyer WM, Egalka MC, Guinan EC: Suppression of allogeneic T-cell proliferation by human marrow stromal cells: implications in transplantation. Transplantation. 2003, 75: 389-97. 10.1097/01.TP.0000045055.63901.A9.

Maccario R, Podestà M, Moretta A, Cometa A, Comoli P, Montagna D, Daudt L, Ibatici A, Piaggio G, Pozzi S, Frassoni F, Locatelli F: Interaction of human mesenchymal stem cells with cells involved in alloantigen-specific immune response favors the differentiation of CD4+ T-cell subsets expressing a regulatory/suppressive phenotype. Haematologic. 2005, 90: 516-25.

Prevosto C, Zancolli M, Canevali P, Zocchi MR, Poggi A: Generation of CD4+ or CD8+ regulatory T cells upon mesenchymal stem cell-lymphocyte interaction. Haematologica. 2007, 92: 881-88. 10.3324/haematol.11240.

Radley HG, De Luca A, Lynch GS, Grounds MD: Duchenne muscular dystrophy: focus on pharmaceutical and nutritional interventions. Int J Biochem Cell Biol. 2007, 39: 469-77. 10.1016/j.biocel.2006.09.009.

Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, Deans R, Keating A, Prockop Dj, Horwitz E: Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy. 2006, 8: 315-7. 10.1080/14653240600855905.

Nguyen F, Cherel Y, Guigand L, Goubault-Leroux I, Wyers M: Muscle lesions associated with dystrophin deficiency in neonatal golden retriever puppies. Comp Pathol. 2002, 126: 100-08. 10.1053/jcpa.2001.0526.

Ambrosio C, Zucconi E, Martins D, Vanucchi C, Perez M, Vieira N, Valadares M, Jazedje T, Miglino M, Zatz M: Extreme clinical variability in GRMD: From neonatal death to asymptomatic carriers. [abstract]. Neuromuscular Disorders. 2007, 17: 776-10.1016/j.nmd.2007.06.056. doi:10.1016/j.nmd.2007.06.056

Tögel F, Westenfelder C: Adult Bone Marrow-Derived Stem Cells for Organ Regeneration and Repair. Developmental Dynamics. 2007, 236: 3321-3331. 10.1002/dvdy.21258.

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