Received for publication May 13, 2005.
Revised July 18, 2005.
Accepted for publication July 19, 2005.

Interaction with Blood Components Plays a Crucial Role in Asialoglycoprotein Receptor-mediated in Vivo Gene Transfer by Galactosylated Lipoplex

Abstract

In this study, we evaluated the effect of blood components (whole blood and serum) on asialoglycoprotein receptor-mediated in vivo gene transfer. The hepatic transfection activity of galactosylated lipoplex pre-incubated with serum was about 10-times higher than that without incubation after intraportal injection in mice. However, pre-incubation with whole blood significantly reduced hepatic transfection activity. Fluorescent resonance energy transfer analysis and agarose gel electrophoresis revealed that pre-incubation with serum reduced the degree of destabilization of the galactosylated lipoplex in blood, partially supporting enhanced hepatic transfection activity by pre-incubation with serum. Inhibition of hepatic transfection activity by pre-dosing galactosylated bovine serum albumin indicated that the galactosylated lipoplex exposed to serum is recognized by asialoglycoprotein-receptors on hepatocytes. Inactivation of serum prior to mixing with galactosylated lipoplex reduced liver accumulation and completely abolished enhancement of hepatic transfection activity by pre-incubation with active serum, suggesting that not only the stability of the lipoplex in blood but also the serum opsonin activity play important roles. Alternatively, pre-incubation with inactivated serum reduced the lung accumulation and inflammatory cytokine production of galactosylated lipoplex. The information provided by this study will be valuable for the future use, design, and development of galactosylated lipoplex for in vivo asialoglycoprotein receptor-mediated gene transfer.

Key words: Drug delivery system, Gene delivery, Gene therapy, Liposomes, Liver, Pharmacokinetics

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