Titre :
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Overexpression of the myostatin antagonist follistatin in normal myoblasts genetically modified with a lentivirus (abstract : congrès international de Myologie, 2005)
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contenu dans :
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Auteurs :
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Congrès international de myologie 2005 (International Congress of Myology 2005; 9-13 mai 2005; Nantes, France) ;
Benabdallah BF ;
Rousseau J ;
Bouchentouf M ;
Tremblay J
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Type de document :
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Article
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Année de publication :
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2005
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Pages :
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p. 112
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Langues:
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Anglais
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Mots-clés :
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colloque
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dystrophine
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follistatine
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immunocytochimie
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muscle squelettique
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myoblaste
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myostatine
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régénération musculaire
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thérapie cellulaire
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traumatisme musculaire
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Résumé :
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Communication n° 172. Background : Duchenne muscular dystrophy is a severe myopathy caused by the absence of a functional dystrophin in muscles. Transplantation of normal myoblasts is a potential therapy that permits to restore the expression of the dystrophin in transplanted muscles by the formation of hybrid dystrophin positive fibers. However, the success of this approach is compromised by the limited regeneration of damaged muscle. Myostatin, the most powerful inhibitor of muscle growth identified to date, is regulated by different antagonist proteins such as follistatin. Our purpose is to block the myostatin signal in mdx and SCID host mice by the respective transplantation of normal murine and human myoblasts genetically modified with a follistatin lentivirus. Methods : 293T packaging cell line was used to produce a lentivirus coding for the short form of follistatin protein under the control of a cytomegalovirus promoter. Normal murine and human myoblasts were infected with the lentivirus to overproduce the follistatin protein. The surexpression of the follistatin in both transfected 293T cells and infected murine and human normal myoblasts was verified by immuno-cytochemistry assay and Western blot. A lentivirus containing the eGFP gene was used as a control. The differentiation of control and lentivirus-follistatin infected myoblasts was also evaluated by the determination of the fusion index. Results : The immunocytochemistry assay showed that the follistatin was clearly overexpressed in both pCMV-Fst transfected 293T cells and infected murine and human normal myoblasts compared with the control cells transfected or infected with the eGFP lentivirus. The Western blot also demonstrated that the follistatin was effectively overexpressed in pCMV-Fst transfected 293T cells and infected murine and human normal myoblasts compared with the control cells transfected or infected with the eGFP lentivirus. The fusion index test also showed that myoblasts infected with the follistatin lentivirus formed more myotubes than myoblasts infected with the eGFP-lentivirus when cultured in differentiation conditions. Conclusions : The transplantation of normal murine or human myoblasts overexpressing follistatin, in mdx or SCID mouse respectively in order to antagonize the myostatin signaling, could be a good approach to improve the success of the potential cellular therapy of Duchenne myopathy.
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