Yoshikazu Maeda, Uichi Ikeda, Masahisa Shimpo, Shuichi Ueno, Yoji Ogasawara, Masashi Urabe, Akihiro Kume, Toshihiro Takizawa, Takuma Saito, Peter Colosi, Gary Kurtzman, Kazuyuki Shimada, Keiya Ozawa
Journal of Molecular and Cellular Cardiology 30(7) 1341-1348 1998年 査読有り
Adeno-associated virus (AAV) vectors, derived from a non-pathogenic parvovirus, are considered to be an appropriate gene transfer vehicle for both dividing and non-dividing cells. In the present study, we investigated whether the rat heart could be efficiently transduced with AAV vectors. Rat cardiac myocytes (CM) were infected with AAV-lacZ vector containing β-galactosidase (β-gal) gene in vitro and the expression of β-gal in CM was evaluated by X-gal staining and β-gal ELISA. With increasing multiplicities of infection (MOI), more than 60% of CM were stained positively with X-gal, and the β-gal expression increased to 31.1 ± 4.6 ng/mg protein in a MOI-dependent manner (MOI: 104 to 106 particles/cell). The β-gal expression was also increased in an incubation period-dependent manner (1-24 h). β-gal expression was maximal at day 3 and then gradually decreased with time. However, β-gal expression at day 14 was almost the same level as that at day 1 (45.5 ± 5.9 v 55.2 ± 6.2 ng/mg protein). Excised rat right ventricular papillary muscles were also infected with AAV-lacZ ex vivo. When the β-gal expression was evaluated by X-gal staining, more than 80% of CM in the papillary muscles were stained positively, indicating efficient gene transfer into CM using AAV vectors. These findings suggest that AAV vectors are promising for cardiac gene therapy.
