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South African Journal of Science

On-line version ISSN 1996-7489
Print version ISSN 0038-2353

S. Afr. j. sci. vol.103 n.5-6 Pretoria May./Jun. 2007




Targeting of glycosylated lipoplexes in HepG2 cells: Anomeric and C-4 epimeric preference of the asialoglycoprotein receptor



Moganavelli SinghI; Colin B. RogersII; Mario AriattiI

IDepartment of Biochemistry, Westville Campus, University of KwaZulu-Natal, Private Bag X54001, Durban 4000, South Africa
IIDepartment of Chemistry, Westville Campus, University of KwaZulu-Natal, Private Bag X54001, Durban 4000, South Africa




This study was conducted to determine the capacity of the asialoglycoprotein receptor on the hepatocyte-derived cell line HepG2 to exhibit an anomeric preference with respect to the D-galacto moiety on cationic liposome membrane-anchored cholesteryl-α-D-galactopyranoside (Choagal) and cholesteryl-ß-D-galactopyranoside (Cholßgal) in cationic liposome/pGL3 plasmid DNA complexes constructed for non-viral, hepatocyte-directed gene transfer. In addition, cholesteryl-α-D-glucopyranoside (Choaglu) and cholesteryl-ß-D-glucopyranoside (Cholßglu) were separately formulated into cationic liposomes at the same mole ratio (11%) to examine the C-4 epimeric selectivity of the asialo-glycoprotein lectin for the glucopyranosides displayed in derived lipoplexes. Lipoplex formation was examined by gel retardation, ethidium displacement assays, and transmission electron microscopy. Plasmid DNA was shown to be fully liposome associated and maximally compacted at a liposome:DNA ratio of 6:1 (weight ratio), corresponding to a +/- charge ratio of 1.3 with complexes falling in the 80-200 nm size range, whereas at a 5:1 w/wratio [1.1 (+/-) charge ratio] lipoplexes were somewhat smaller (50-100 nm) but promoted higher transgene activity in HepG2 cells than 6:1 (w/w) lipoplexes, in the following order: Cholßgal>Cholαgal > Cholßglu = Cholαglu. Transgene activity levels in HeLa cells which lack the asialoglycoprotein receptor were approximately 10% of those achieved in HepG2 cells. Moreover, transgene activity in HepG2 cells was reduced by approximately 90% in the presence of excess asialofetuin, a ligand for the asialoglycoprotein lectin.



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Received 23 March 2007.
Accepted 22 June 2007.



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