| Research Abstract |
Goal of this study is development of new gene and cell therapy approach for neuro-muscular disorders. Especially we focus on muscular dystrophies and try to establish dystrophin replacement therapy. 1) We have generated series of viral vectors for gene delivery : containing maker genes, LacZ and eGFP, various short version of dystrophin with or without eGFP fusion gene, skeletal muscle specific transcriptional factors, MyoD, Pax3, and Pax7. To drive these genes of interest, we have also prepared series of promoter sets, ubiquitous promoters : CMV, RSV, and CAG, and skeletal muscle specific promoters : Human Skeletal a-actin gene promoter (HSA), CK6 (modified Creatinin Kinase promoter version 6), and MHCK7. 2) We successfully presented 'cell-fate control therapy model' of DMD : Fibroblasts derived from mdx mouse tail were trasnduced with a myoD-ER(T) gene, which is regulatable myoD, then myo-converted with Tamoxifen ex vivo as well as in vivo, to have 'new muscle cells' to supply a therapeutic micro-dystrophin protein in mdx mouse muscles (19. Kimura et al 2008). 3) We have also reported at least two year period of dystrophin gene expression after in vivo direct delivery with lentiviral vector into mdx mouse muscle, In this case, we demonstrated pathological and physiological function were recovered after dystrophin gene replacement and delivered gene were stably exist in muscle satellite cells which were muscle specific stem cells (11. Kimura et al 2010). 4) We have also generated Lv-mediated transgenic mouse to analyze promoter cassettes for therapeutic gene expression. In these animals, we have checked gene expression levels in various tissues such as skeletal muscle, heart, and others, also shown fiber type variations of gene expression in skeletal muscles (2. Suga et al 2011).
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