Clinical and basic scientific researchi toward gene therapy in progressive muscular dystrophies

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We have conducted the molecular genetic analysis and clinical application for genotype-phenotype correlation in progressive muscular dystrophies, especially Duchenne muscular dystrophy (DMD) and Fukuyama congenital muscular dystrophy (FCMD). For the purpose of obtaining complinentary primary genetic defects in the DMD gene by introducing copies of recombinant gene constructs into nuscle cells ex vivo, the potential use of cationic liposomes as physical gene delivery systems for cultured human dystrophic skeletal muscles was examined, from the basic scientific point of view. Various cationic liposome formulations, i.e.cellfectin, DMRIE-C,lipofectin, lipofectAMINE and lipofectAMINE PLUS,were examined for their use in plasmid DNA (beta-gal reporter gene) transfection. For transgene expression, muscle cultures were stained or were applied to detect beta-gal activities by the ELISA method. These in vitro studies indicated that lipofectAMINE PLUS showed high efficiency. In addition, myoblast non-clonal cultures were more efficient than clonal cultures. This is the first successful introduction of foreign genes into primary cultures of human dystrophic myoblasts by means of cationic liposomes.