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KMID : 0617320020110010293
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Journal of Pharmacetical Sceiences Ewha Womans University
2002 Volume.11 No. 1 p.293 ~ p.304
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Receptor-mediated delivery of an antisense gene to human brain cancer cells
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Zhang, Yun
Lee, Hwa Jeong/J. Boado, Ruben/M. Pardridge, William
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Abstract
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Background The goal of this work was the development of a gene targetingtechnology that will enable the delivery of therapeutic genes to brain cancercells in vivo following intravenous administration. High-grade brain gliomasoverexpress the epidermal growth factor receptor (EGFR) and EGFR anti-sense gene therapy could reduce the growth of EGFR-dependent gliomas.
Methods A human EGFR antisense gene driven by the SV4O promoter ina non-virat plasmid carrying elements that facilitate extra-chromosomalreplication was packaged in the interior of 85 nm pegylated immuno-liposomes (PILs) . The PILs were targeted to U87 human glioma cells withthe 83-14 murine monoclonal antibody (MAb) to the human insulin receptor(HIR).
Results Confocal fluorescent microscopy demonstrated that the unconju-gated HIR MAb is rapidly internalized by the glioma cells. Endoytosisfellowed by entry into the nucleus was also demonstrated for the HIR MAbconjugated PILs carrying fluorescein-labeled plasmid DNA. The PILs deliveredenogenous genes to virtually all cells in culture, based on β-galactosidasehistochemistry. The targeting of a luciferasr gene to the U87 cells with thePILs resulted in luciferase levels in excess of 150 pg/mg protein after 72 h ofincubation. The level of luciferase gene expression in the U87 cells acHievedwith the PIL gene targeting system was comparable to that with lipo-fectamine. Targeting the EGFR antisense gene to U87 glioma cells with thePILs resulted in more than 70% reduction in [^3H]thymidine incorporationinto the cells; this was paralleled by a 79% reduction in the level ofimmunoreactive EGFR.
Conclusion The present work describes the targeting of an EGFR antisensegene to human brain cancer cells, which results in a 70-80% inhibition incancer cell growth. PILs provide a new approach to gene targeting that iseffective in vivo following intravenous administration without viral vectors.
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