Enhancing effects of galactosylated dendrimer/alpha-cyclodextrin conjugates on gene transfer efficiency.
To improve in vitro gene transfer efficiency and/or achieve cell-specific gene delivery of polyamidoamine (PAMAM) starburst dendrimer (generation 2, G2) conjugate with alpha-cyclodextrin (alpha-CDE conjugate (G2)), we prepared alpha-CDE conjugate bearing galactose (Gal-alpha-CDE conjugates) with the various degrees of substitution of the galactose moiety (DSG) as a novel non-viral vector. The agarose gel electrophoretic studies revealed that Gal-alpha-CDE conjugates formed complexes with plasmid DNA (pDNA) and protected the degradation of pDNA by DNase I, but these effects impaired as the DSG value increased. Dendrimer and alpha-CDE conjugate exerted pDNA condensation through the complexation, but Gal-alpha-CDE conjugates did not. Gal-alpha-CDE conjugate (DSG 4) was found to have much higher gene transfer activity than dendrimer, alpha-CDE conjugate and Gal-alpha-CDE conjugates (DSG 8, 15) in HepG2, NIH3T3 and A549 cells, which are independent of the expression of the asialoglycoprotein receptor. Transfection activity of Gal-alpha-CDE conjugate (DSG 4) was insensitive to the existence of competitors (asialofetuin and galactose) and serum. In addition, no cytotoxicity after transfection of the complex of pDNA with Gal-alpha-CDE conjugate (DSG 4) was observed. These results suggest the potential use of Gal-alpha-CDE conjugate (DSG 4) as a non-viral vector in various cells.
Wada
Koki
K
Graduate School of Pharmaceutical Sciences, Kumamoto University, Oe-honmachi, Kumamoto, Japan.
Arima
Hidetoshi
H
Tsutsumi
Toshihito
T
Hirayama
Fumitoshi
F
Uekama
Kaneto
K
D000818:Animals; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D005690:Galactose / Q000008:administration & dosage* / Q000737:chemistry / Q000378:metabolism*; D018014:Gene Transfer Techniques; D006801:Humans; D051379:Mice; D041681:NIH 3T3 Cells; D047391:alpha-Cyclodextrins / Q000008:administration & dosage* / Q000737:chemistry / Q000378:metabolism*
D047391:alpha-Cyclodextrins; D005690:Galactose; C032613:alpha-cyclodextrin