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Herpes simplex virus vector‐mediated dystrophin gene transfer and expression in MDX mouse skeletal muscle
Author(s) -
Akkaraju Giridhar R.,
Huard Johnny,
Hoffman Eric P.,
Goins William F.,
Pruchnic Ryan,
Watkins Simon C.,
Cohen Justus B.,
Glorioso Joseph C.
Publication year - 1999
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/(sici)1521-2254(199907/08)1:4<280::aid-jgm45>3.0.co;2-l
Subject(s) - gene transfer , herpes simplex virus , virology , vector (molecular biology) , dystrophin , mdx mouse , skeletal muscle , biology , viral vector , gene , virus , anatomy , genetics , recombinant dna
Background Duchenne muscular dystrophy (DMD) results from mutations that prevent the expression of functional dystrophin in muscle fibers. Herpes simplex virus type‐1 (HSV‐1) represents a potentially useful vector for treatment of DMD because it has the capacity to accommodate the 14‐kb full‐length dystrophin cDNA and can efficiently transduce muscle cells. We have tested the ability of first‐ and second‐generation replication‐defective HSV vectors to deliver full‐length dystrophin to dystrophin‐deficient mdx muscle cells in vitro and in vivo . Methods First‐generation replication‐defective HSV vectors harboring full‐length or truncated (Becker) dystrophin expression cassettes and lacking a single viral immediate‐early (IE) gene were constructed and tested by immunofluorescence and immunoblotting for their ability to direct dystrophin expression in infected mdx cells in culture. To reduce vector cytotoxicity and safety concerns, a second‐generation dystrophin vector missing additional IE genes was constructed and tested in vitro and in vivo . Results Dystrophin expression was observed in infected mdx myotubes in vitro in all cases. Confocal microscopy showed exclusive localization of full‐length dystrophin to the cell membrane whereas the Becker variant was also found abundantly throughout the cytoplasm. Dystrophin expression in mdx mice was restored in muscle cells near the site of vector injection. Conclusion Highly defective HSV‐1 vectors which lack the ability to spread systemically and are greatly reduced in toxicity for infected cells, thus removing an impediment to prolonged transgene expression, can direct the delivery and proper expression of full‐length dystrophin whose considerable size is compatible with few other modes of delivery. These vectors may offer a legitimate opportunity toward the development of effective gene therapy treatments for DMD. Copyright © 1999 John Wiley & Sons, Ltd.