title: Sendai virus-mediated gene delivery into hepatocytes via isolated hepatic perfusion.
abstract: 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.
authors.affiliation: Department of Surgery, E1, Osaka University Graduate School of Medicine, Japan.
authors.forename: Shigeo
authors.initials: S
authors.lastname: Fujita
authors.forename: Akiko
authors.initials: A
authors.lastname: Eguchi
authors.forename: Jun
authors.initials: J
authors.lastname: Okabe
authors.forename: Atsushi
authors.initials: A
authors.lastname: Harada
authors.forename: Katsunori
authors.initials: K
authors.lastname: Sasaki
authors.forename: Naoko
authors.initials: N
authors.lastname: Ogiwara
authors.forename: Yoshifumi
authors.initials: Y
authors.lastname: Inoue
authors.forename: Toshinori
authors.initials: T
authors.lastname: Ito
authors.forename: Hikaru
authors.initials: H
authors.lastname: Matsuda
authors.forename: Kazunori
authors.initials: K
authors.lastname: Kataoka
authors.forename: Atsushi
authors.initials: A
authors.lastname: Kato
authors.forename: Mamoru
authors.initials: M
authors.lastname: Hasegawa
authors.forename: Mahito
authors.initials: M
authors.lastname: Nakanishi
mesh_terms: D000818:Animals; D005822:Genetic Vectors; D022781:Hepatocytes / Q000378:metabolism*; D008297:Male; D010477:Perfusion; D051381:Rats; D017208:Rats, Wistar; D029082:Sendai virus / Q000235:genetics*