title: Analysis of muscle creatine kinase regulatory elements in recombinant adenoviral vectors.
abstract: Adenoviral gene transfer holds promise for gene therapy, but effective transduction of a large and distributed tissue such as muscle will almost certainly require systemic delivery. In this context, the use of muscle-specific regulatory elements such as the muscle creatine kinase (MCK) promoter and enhancer will avoid potentially harmful ectopic expression of transgenes. We describe here the development and testing of adenoviral vectors containing small, striated muscle-specific, highly active MCK expression cassettes. One of these regulatory elements (CK6) is less than 600 bp in length and is 12% as active as the CMV promoter/enhancer in muscle. A recombinant adenoviral vector containing this regulatory element retains very high muscle specificity, expressing 600-fold higher levels of transgene in muscle than in liver. Muscle-specific regulatory elements may also increase persistence of transduced muscle cells. Adenoviral transduction of dendritic cells has been shown to stimulate cytotoxic T-lymphocyte (CTL) responses directed against transgene epitopes. We show that human dendritic cells infected in vitro with MCK-containing adenoviruses do not express significant levels of transgene. Furthermore, while adenoviral vectors containing nonspecific promoters are normally cleared from muscle tissue within 1 month, we show that MCK-containing vectors express significant levels of transgene 4 months after intramuscular injection.
authors.affiliation: Department of Human Genetics and Center for Gene Therapy, University ofMichigan, Ann Arbor, MI 48109, USA.
authors.forename: M A
authors.initials: MA
authors.lastname: Hauser
authors.forename: A
authors.initials: A
authors.lastname: Robinson
authors.forename: D
authors.initials: D
authors.lastname: Hartigan-O'Connor
authors.forename: D A
authors.initials: DA
authors.lastname: Williams-Gregory
authors.forename: J N
authors.initials: JN
authors.lastname: Buskin
authors.forename: S
authors.initials: S
authors.lastname: Apone
authors.forename: C J
authors.initials: CJ
authors.lastname: Kirk
authors.forename: S
authors.initials: S
authors.lastname: Hardy
authors.forename: S D
authors.initials: SD
authors.lastname: Hauschka
authors.forename: J S
authors.initials: JS
authors.lastname: Chamberlain
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003402:Creatine Kinase / Q000235:genetics*; D003713:Dendritic Cells / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000493:pharmacokinetics; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology; D008099:Liver / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009132:Muscles / Q000201:enzymology* / Q000378:metabolism; D016296:Mutagenesis; D011401:Promoter Regions, Genetic; D012045:Regulatory Sequences, Nucleic Acid; D013997:Time Factors; D019076:Transgenes; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D003402:Creatine Kinase; D001616:beta-Galactosidase