title: Gene transfer into vascular cells using adeno-associated virus (AAV) vectors.
abstract: OBJECTIVE
  Recombinant viral vectors based on the nonpathogenic parvovirus, adeno-associated virus (AAV), have a number of attractive features for gene therapy, including the ability to transduce non-dividing cells and its long-term transgene expression. In this study, an AAV vector containing bacterial beta-galactosidase gene (lacZ) was used to transduce cultured rat vascular smooth muscle cells (VSMC) in vitro and rat thoracic aortas ex vivo.
  
  
  METHODS
  VSMC were transduced with AAV-lacZ at multiplicities of infection (MOI) ranging from 5.0 x 10(5) to 1.0 x 10(7). Expression of beta-galactosidase (beta-gal) in VSMC was evaluated by X-gal staining and a beta-gal ELISA method. Excised rat aortas were incubated with medium containing AAV-lacZ. Expression of beta-gal in the aortic segments was evaluated by X-gal staining.
  
  
  RESULTS
  With increasing MOI, up to 50% of cultured VSMC were positive by X-gal staining and the beta-gal expression increased up to 15 ng/mg protein. The expression gradually decreased during the culture but was detectable for at least 1 month. In the ex vivo study, AAV vectors transduced endothelial and adventitial cells in rat aortic segments, while no expression was seen in medial VSMC.
  
  
  CONCLUSIONS
  AAV vectors can efficiently transduce rat VSMC in vitro. AAV-mediated ex vivo gene transfer into the normal aorta resulted in efficient gene transfer into endothelial and adventitial cells but not into medial VSMC. These findings suggest that AAV-based vectors are promising for use in cardiovascular gene therapy.
authors.affiliation: Department of Cardiology, Jichi Medical School, Tochigi, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Maeda
authors.forename: U
authors.initials: U
authors.lastname: Ikeda
authors.forename: Y
authors.initials: Y
authors.lastname: Ogasawara
authors.forename: M
authors.initials: M
authors.lastname: Urabe
authors.forename: T
authors.initials: T
authors.lastname: Takizawa
authors.forename: T
authors.initials: T
authors.lastname: Saito
authors.forename: P
authors.initials: P
authors.lastname: Colosi
authors.forename: G
authors.initials: G
authors.lastname: Kurtzman
authors.forename: K
authors.initials: K
authors.lastname: Shimada
authors.forename: K
authors.initials: K
authors.lastname: Ozawa
mesh_terms: D001013:Aorta, Thoracic / Q000166:cytology; D002478:Cells, Cultured; D000229:Dependovirus; D004730:Endothelium, Vascular / Q000166:cytology; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D007763:Lac Operon; D009131:Muscle, Smooth, Vascular / Q000166:cytology*