title: In vitro and in vivo gene delivery mediated by Lactosylated dendrimer/alpha-cyclodextrin conjugates (G2) into hepatocytes.
abstract: The purpose of this study is to evaluate in vitro and in vivo gene delivery efficiency of polyamidoamine (PAMAM) starburst dendrimer (generation 2, G2) conjugates with alpha-cyclodextrin (alpha-CDE (G2)) bearing lactose (Lac-alpha-CDE) with various degrees of substitution of the lactose moiety (DSL) as a novel hepatocyte-selective carrier in hepatocytes. Lac-alpha-CDE (DSL 2.6) was found to have much higher gene transfer activity than dendrimer, alpha-CDE, Lac-alpha-CDE (DSL 1.2, 4.6, 6.2 and 10.2) and lactosylated dendrimer (Lac-dendrimer, DSL 2.4) in HepG2 cells, which are dependent on the expression of cell-surface asialoglycoprotein receptor (ASGP-R), reflecting the cellular association of the plasmid DNA (pDNA) complexes. The physicochemical properties of pDNA complex with Lac-alpha-CDE (DSL 2.6) were almost comparable to that with alpha-CDE. Lac-alpha-CDE (DSL 2.6) provided negligible cytotoxicity up to a charge ratio of 150 in HepG2 cells. Lac-alpha-CDE (DSL 2.6) provided gene transfer activity higher than jetPEI-Hepatocyte to hepatocytes with much less changes of blood chemistry values 12h after intravenous administration in mice. These results suggest the potential use of Lac-alpha-CDE (DSL 2.6) as a non-viral vector for gene delivery toward hepatocytes.
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kumamoto University, 5-1 Oe-honmachi, Kumamoto 862-0973, Japan. arimah@gpo.kumamoto-u.ac.jp
authors.forename: Hidetoshi
authors.initials: H
authors.lastname: Arima
authors.forename: Shogo
authors.initials: S
authors.lastname: Yamashita
authors.forename: Yoshimasa
authors.initials: Y
authors.lastname: Mori
authors.forename: Yuya
authors.initials: Y
authors.lastname: Hayashi
authors.forename: Keiichi
authors.initials: K
authors.lastname: Motoyama
authors.forename: Kenjiro
authors.initials: K
authors.lastname: Hattori
authors.forename: Tomoko
authors.initials: T
authors.lastname: Takeuchi
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Jono
authors.forename: Yukio
authors.initials: Y
authors.lastname: Ando
authors.forename: Fumitoshi
authors.initials: F
authors.lastname: Hirayama
authors.forename: Kaneto
authors.initials: K
authors.lastname: Uekama
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D004247:DNA / Q000008:administration & dosage / Q000235:genetics*; D050091:Dendrimers / Q000737:chemistry*; D004337:Drug Carriers / Q000737:chemistry*; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D007624:KB Cells; D007785:Lactose / Q000737:chemistry*; D008099:Liver / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D018613:Microscopy, Confocal; D009928:Organ Specificity; D010957:Plasmids; D011094:Polyethyleneimine / Q000737:chemistry; D020349:Surface Plasmon Resonance; D047391:alpha-Cyclodextrins / Q000737:chemistry*
chemical_list: D050091:Dendrimers; D004337:Drug Carriers; C104700:PAMAM Starburst; D047391:alpha-Cyclodextrins; D011094:Polyethyleneimine; D004247:DNA; D007785:Lactose