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J. Biol. Chem., Vol. 276, Issue 36, 33875-33880, September 7, 2001

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Extracellular Glycosaminoglycans Modify Cellular Trafficking of Lipoplexes and Polyplexes^*

Marika Ruponen^§, Seppo Rönkkö, Paavo Honkakoski, Jukka Pelkonen^¶, Markku Tammi^**, and Arto Urtti

From the Departments of  Pharmaceutics, ^¶ Clinical Microbiology, and ^** Anatomy, University of Kuopio and the  Department of Pediatrics, Kuopio University Hospital, FIN-70211 Kuopio, Finland

It has been shown that extracellular glycosaminoglycans (GAGs) limit the gene transfer by cationic lipids and polymers. The purpose of this study was to clarify how interactions with anionic GAGs (hyaluronic acid and heparan sulfate) modify the cellular uptake and distribution of lipoplexes and polyplexes. Experiments on cellular DNA uptake and GFP reporter gene expression showed that decreased gene expression can rarely be explained by lower cellular uptake. In most cases, the cellular uptake is not changed by GAG binding to the lipoplexes or polyplexes. Reporter gene expression is decreased or blocked by heparan sulfate, but it is increased by hyaluronic acid; this suggests that intracellular factors are involved. Confocal microscopy experiments demonstrated that extracellular heparan sulfate and hyaluronic acid are taken into cells both with free and DNA-associated carriers. We conclude that extracellular GAGs may alter both the cellular uptake and the intracellular behavior of the DNA complexes.

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