The potential role of proteoglycans in cationic lipid-mediated gene delivery. Studies of the interaction of cationic lipid-DNA complexes with model glycosaminoglycans.
Recent evidence supports a role for proteoglycans in polycation-mediated gene delivery. Therefore, the interaction of glycosaminoglycans with cationic lipid-DNA complexes (CLDCs) has been characterized using a combination of biophysical approaches. At low ionic strength, CLDCs bind to heparin-derivatized Sepharose particles, with the ratio of cationic lipid to DNA controlling the binding. Incorporation of the helper lipids cholesterol or 1,2-dioleoyl-phosphatidylethanolamine increases the amount of bound CLDC. Heparin also induces the aggregation of CLDCs, with cholesterol reducing this effect. Isothermal titration calorimetry demonstrates an endothermic heat for the binding of heparin to CLDCs at low ionic strength, whereas circular dichroism studies suggest a heparin-stimulated release of DNA from CLDCs at a greater than 20-fold charge excess. Increasing the ionic strength to 0.11 reduces CLDC binding to heparin beads, and greatly enhances the release of DNA from CLDCs by heparin. The ability of the cell surface glycosaminoglycan heparan sulfate to release DNA from CLDCs is more sensitive than heparin to the incorporation of the cholesterol or 1,2-dioleoyl-phosphatidylethanolamine. Titration calorimetry reveals an exothermic heat for the interaction glycosaminoglycans with CLDCs at higher ionic strength. These results are consistent with the direct involvement of proteoglycans in transfection.
Wiethoff
C M
CM
Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, Kansas 66047, USA.
Smith
J G
JG
Koe
G S
GS
Middaugh
C R
CR
D002151:Calorimetry; D004247:DNA / Q000737:chemistry*; D004337:Drug Carriers; D005229:Fatty Acids, Monounsaturated / Q000737:chemistry*; D018014:Gene Transfer Techniques; D020404:Glycerophospholipids / Q000737:chemistry*; D006025:Glycosaminoglycans / Q000737:chemistry*; D006493:Heparin / Q000737:chemistry*; D007700:Kinetics; D008027:Light; D008081:Liposomes; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds / Q000737:chemistry*; D012542:Scattering, Radiation; D013816:Thermodynamics
C410262:1,2-dioleoyl-glycero-3-phosphatidyl ethanolamine; D004337:Drug Carriers; D005229:Fatty Acids, Monounsaturated; D020404:Glycerophospholipids; D006025:Glycosaminoglycans; D008081:Liposomes; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds; D006493:Heparin; D004247:DNA; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane