Efficient gene transfer into cardiac myocytes using adeno-associated virus (AAV) vectors.
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 beta-galactosidase (beta-gal) gene in vitro, and the expression of beta-gal in CM was evaluated by X-gal staining and beta-gal ELISA. With increasing multiplicities of infection (MOI), more than 60% of CM were stained positively with X-gal, and the beta-gal expression increased to 31.1 +/- 4.6 ng/mg protein in a MOI-dependent manner (MOI: 10(4) to 10(6) particles/cell). The beta-gal expression was also increased in an incubation period-dependent manner (1-24 h). beta-gal expression was maximal at day 3 and then gradually decreased with time. However, beta-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 beta-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.
Maeda
Y
Y
Department of Cardiology, Jichi Medical School, Tochigi, Japan.
Ikeda
U
U
Shimpo
M
M
Ueno
S
S
Ogasawara
Y
Y
Urabe
M
M
Kume
A
A
Takizawa
T
T
Saito
T
T
Colosi
P
P
Kurtzman
G
G
Shimada
K
K
Ozawa
K
K
D000818:Animals; D002478:Cells, Cultured; D000229:Dependovirus; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors; D006352:Heart Ventricles / Q000166:cytology; D009206:Myocardium / Q000166:cytology*; D010210:Papillary Muscles / Q000166:cytology; D051381:Rats; D017207:Rats, Sprague-Dawley; D014161:Transduction, Genetic; D001616:beta-Galactosidase / Q000235:genetics
D001616:beta-Galactosidase