Sendai virus-mediated gene delivery into hepatocytes via isolated hepatic perfusion.
The recombinant Sendai virus vector is a promising tool for human gene therapy, capable of inducing high-level expression of therapeutic genes in tissue cells in situ. The target tissues include airway epithelium, blood vessels, skeletal muscle, retina and the central nervous system, but application to hepatic tissues has not yet been achieved, because direct intraportal injection of the vector is not feasible. We report an efficient and harmless procedure of gene delivery by recombinant Sendai virus into rat parenchymal hepatocytes, based on isolated hepatic perfusion with controlled inflow. Critical parameters for successful hepatic gene delivery are a brief preperfusion period (25 degrees C, 5 min); appropriate vector concentration in the perfusate (10(7) pfu/ml); moderate portal vein pressure (12 mmHg) and a brief hyperthermic postperfusion period (42 degrees C, 5 min). Under these optimized conditions, marker genes were expressed in most parenchymal hepatocytes without significant damage to hepatic tissues. Furthermore, expression of the marker genes was undetectable in nonhepatic tissues, including the gonads, indicating that this approach strictly targets hepatic tissues and thus offers good clinical potential for human gene therapy.
Fujita
Shigeo
S
Department of Surgery, E1, Osaka University Graduate School of Medicine, Japan.
Eguchi
Akiko
A
Okabe
Jun
J
Harada
Atsushi
A
Sasaki
Katsunori
K
Ogiwara
Naoko
N
Inoue
Yoshifumi
Y
Ito
Toshinori
T
Matsuda
Hikaru
H
Kataoka
Kazunori
K
Kato
Atsushi
A
Hasegawa
Mamoru
M
Nakanishi
Mahito
M
D000818:Animals; D005822:Genetic Vectors; D022781:Hepatocytes / Q000378:metabolism*; D008297:Male; D010477:Perfusion; D051381:Rats; D017208:Rats, Wistar; D029082:Sendai virus / Q000235:genetics*