Effect of serum components on the physico-chemical properties of cationic lipid/oligonucleotide complexes and on their interactions with cells
| Autor: | Lisa S. Uyechi, Francis C. Szoka, Olivier Zelphati, Lee G. Barron |
|---|---|
| Rok vydání: | 1998 |
| Předmět: |
Biophysics
Oligonucleotides Biochemistry Immunoglobulin G Fatty Acids Monounsaturated Endocrinology Cationic liposome Bovine serum albumin Cells Cultured Serum Albumin biology Chemistry Heparin Phosphatidylethanolamines Cell Membrane Cationic polymerization hemic and immune systems Blood Proteins respiratory system Lipid Metabolism Blood proteins Lipids In vitro Macroglobulin Complement system Lipoproteins LDL Quaternary Ammonium Compounds Cholesterol Microscopy Fluorescence biology.protein lipids (amino acids peptides and proteins) Lipoproteins HDL Oleic Acid |
| Zdroj: | Biochimica et biophysica acta. 1390(2) |
| ISSN: | 0006-3002 |
| Popis: | The interactions among serum components and cationic lipid–nucleic acid complexes are central to the understanding of how serum inhibits cellular delivery of oligonucleotides in vitro and in vivo. In this study, we show that several serum proteins, in particular bovine serum albumin (BSA), lipoproteins (HDL and LDL) and macroglobulin, interact with cationic lipid/oligonucleotide complexes, alter the complex diameter and zeta potential (from positive to negative values), and significantly interfere with the ability of 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) to deliver phosphorothioate oligonucleotides (ODN) into cells. Serum and BSA do not dissociate the ODN and lipid components, therefore inhibition of delivery cannot be attributed to a displacement of cationic lipid from the ODN. Rather BSA at 2.5 mg/ml, comparable to the amount found in 10% serum, decreases the cell association of ODN by about 5-fold and nuclear uptake of ODN by greater than 20-fold. In contrast, immunoglobulin G, the other major serum component, alters the zeta potential from positive to near neutral, has a modest effect on the diameter of the complex but does not affect cell association or nuclear delivery of the ODN at amounts found in 10% serum. Other molecules found in serum, specifically oleic acid and heparin, displace the ODN from the complex and thus interfere with delivery. This displacement is attenuated by first incubating the complex with BSA. Another manifestation of serum–complex interactions is that ODN significantly and cationic liposomes slightly, activate complement. However, formation of the complex markedly reduces the complement activation of the ODN. Finally, the effect of serum can be partially counteracted by the selection of the helper lipid (DOPE or cholesterol). Inclusion of a helper lipid reduces the effective charge ratio (cationic groups/anionic thioates) required to deliver ODN into cells and permits delivery in the presence of greater percentages of serum in the culture medium. These results support the current view that the binding of serum proteins to the complex is a significant factor in modulating the activity of cationic lipid–ODN complexes in culture and after intravenous administration. |
| Databáze: | OpenAIRE |
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