© 1997 by European Society of Cardiology
Copyright © 1997, European Society of Cardiology
Gene transfer into vascular cells using adeno-associated virus (AAV) vectors
aDepartment of Cardiology, Jichi Medical School, Minamikawachi-machi, Tochigi, Japan
bDepartment of Molecular Biology, Institute of Hematology, Jichi Medical School, Minamikawachi-machi, Tochigi, Japan
cDepartment of Anatomy, Jichi Medical School, Minamikawachi-machi, Tochigi, Japan
dAvigen Inc., Alameda, CA, USA
* Corresponding author. Department of Molecular Biology, Institute of Hematology, Jichi Medical School, Minamikawachi-machi, Tochigi 329-04, Japan. Tel.: +81 (285) 44-2111; fax: +81 (285) 44-8675; e-mail: kozawa@jichi.ac.jp
Objectives: 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 β-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 5.0x105 to 1.0x107. Expression of β-galactosidase (β-gal) in VSMC was evaluated by X-gal staining and a β-gal ELISA method. Excised rat aortas were incubated with medium containing AAV-lacZ. Expression of β-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 β-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.
KEYWORDS Adeno-associated virus vector; Gene transfer; Smooth muscle cell; Endothelial cell; Aorta
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