Analysis of muscle creatine kinase regulatory elements in recombinant adenoviral vectors

Michael A. Hauser, Ann Robinson, Dennis Hartigan-O'Connor, DeeAnn Williams-Gregory, Jean N. Buskin, Steve Apone, Christopher J. Kirk, Stephen Hardy, Stephen D. Hauschka, Jeffrey S. Chamberlain

Research output: Contribution to journalArticle

73 Scopus citations

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 asthe 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 contain ing 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.

Original languageEnglish (US)
Pages (from-to)16-25
Number of pages10
JournalMolecular Therapy
Volume2
Issue number1
DOIs
StatePublished - Jul 2000
Externally publishedYes

Keywords

  • Adenovirus
  • Dendritic cell
  • Muscle creatine kinase
  • Muscle-specific
  • Muscular dystrophy

ASJC Scopus subject areas

  • Molecular Biology

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  • Cite this

    Hauser, M. A., Robinson, A., Hartigan-O'Connor, D., Williams-Gregory, D., Buskin, J. N., Apone, S., Kirk, C. J., Hardy, S., Hauschka, S. D., & Chamberlain, J. S. (2000). Analysis of muscle creatine kinase regulatory elements in recombinant adenoviral vectors. Molecular Therapy, 2(1), 16-25. https://doi.org/10.1006/mthe.2000.0089