title: Effective repetitive dystrophin gene transfer into skeletal muscle of adult mdx mice using a helper-dependent adenovirus vector expressing the coxsackievirus and adenovirus receptor (CAR) and dystrophin.
abstract: BACKGROUND
  The helper-dependent adenovirus (HDAd) vector is less immunogenic and has a larger cloning capacity of up to 37 kb enough to carry the full-length dystrophin cDNA. However, high and long-term expression of dystrophin transduced to mature muscle still remains difficult. One of the main reasons for this is that the expression of the coxsackievirus and adenovirus receptor (CAR) is very low in mature muscle.
  
  
  METHODS
  We have constructed two different HDAd vectors. One contains the LacZ and the murine full-length dystrophin expression cassette (HDAdLacZ-dys), and the other is a new, improved vector containing the CAR and the dystrophin expression cassette (HDAdCAR-dys).
  
  
  RESULTS
  We initially demonstrated high dystrophin expression and prevention of the dystrophic pathology in mdx muscle injected during the neonatal phase with HDAdLacZ-dys. Furthermore, we demonstrated that repeated injections of HDAdCAR-dys into mature muscle led to approximately nine times greater dystrophin-positive fibers in number than a single injection, thereby recovering the expression of dystrophin-associated proteins. This data has also shown that HDAdCAR-dys enabled administration of adenovirus (Ad) vector to the host with pre-existing immunity to the same serotype of Ad.
  
  
  CONCLUSIONS
  Repetitive injections of the HDAd vector containing the CAR and the dystrophin expression cassette could improve the efficiency of subsequent dystrophin gene transfer to mature mdx muscle. This result suggests that our new HDAd vector will provide a novel gene therapy strategy for Duchenne muscular dystrophy, raising the prospects for gene therapy of other hereditary myopathies.
authors.affiliation: Department of Neurology, Graduate School of Medical Sciences, Kumamoto University, 1-1-1 Honjo, Kumamoto 860-0811, Japan.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Uchida
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Maeda
authors.forename: En
authors.initials: E
authors.lastname: Kimura
authors.forename: Satoshi
authors.initials: S
authors.lastname: Yamashita
authors.forename: Yasuto
authors.initials: Y
authors.lastname: Nishida
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Arima
authors.forename: Teruyuki
authors.initials: T
authors.lastname: Hirano
authors.forename: Eiichiro
authors.initials: E
authors.lastname: Uyama
authors.forename: Shuji
authors.initials: S
authors.lastname: Mita
authors.forename: Makoto
authors.initials: M
authors.lastname: Uchino
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D016189:Dystrophin / Q000172:deficiency* / Q000235:genetics* / Q000378:metabolism; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics; D006378:Helper Viruses / Q000235:genetics / Q000502:physiology*; D006801:Humans; D007111:Immunity, Cellular / Q000276:immunology; D007273:Injections, Intramuscular; D051379:Mice; D008810:Mice, Inbred C57BL; D018101:Mice, Inbred mdx; D018482:Muscle, Skeletal / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D020388:Muscular Dystrophy, Duchenne / Q000473:pathology / Q000517:prevention & control; D011991:Receptors, Virus / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins
chemical_list: C568771:CLMP protein, human; C568772:CLMP protein, mouse; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D016189:Dystrophin; D011991:Receptors, Virus; D011993:Recombinant Fusion Proteins