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.
Department of Cardiology, Jichi Medical School, Tochigi, Japan.
Y
Y
Maeda
U
U
Ikeda
M
M
Shimpo
S
S
Ueno
Y
Y
Ogasawara
M
M
Urabe
A
A
Kume
T
T
Takizawa
T
T
Saito
P
P
Colosi
G
G
Kurtzman
K
K
Shimada
K
K
Ozawa
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