1. Delivery of oligoribonucleotides to human hepatoma cells using cationic lipid particles conjugated to ferric protoporphyrin IX (heme).
- Author
-
Takle GB, Thierry AR, Flynn SM, Peng B, White L, Devonish W, Galbraith RA, Goldberg AR, and George ST
- Subjects
- Animals, Cations, Cell Line, Cell Nucleus metabolism, Chlorocebus aethiops, Cytoplasm metabolism, DNA, Recombinant administration & dosage, DNA, Recombinant pharmacokinetics, Drug Carriers, Fatty Acids, Monounsaturated chemistry, Fatty Acids, Monounsaturated pharmacokinetics, Genes, Reporter, Genetic Vectors administration & dosage, Genetic Vectors pharmacokinetics, Heme chemistry, Heme pharmacokinetics, Humans, Kidney, Luciferases biosynthesis, Luciferases genetics, Mice, Microscopy, Fluorescence, Oligoribonucleotides chemistry, Oligoribonucleotides pharmacokinetics, Organ Specificity, Particle Size, Phosphatidylethanolamines chemistry, Phosphatidylethanolamines pharmacokinetics, Quaternary Ammonium Compounds chemistry, Quaternary Ammonium Compounds pharmacokinetics, Recombinant Fusion Proteins biosynthesis, Recombinant Fusion Proteins genetics, Species Specificity, Tumor Cells, Cultured, Vero Cells, Carcinoma, Hepatocellular pathology, Fatty Acids, Monounsaturated administration & dosage, Heme administration & dosage, Liver Neoplasms pathology, Neoplasm Proteins metabolism, Oligoribonucleotides administration & dosage, Phosphatidylethanolamines administration & dosage, Quaternary Ammonium Compounds administration & dosage, Receptors, Cell Surface metabolism
- Abstract
The receptor-ligand interaction between hepatocyte heme receptors and heme was evaluated as a basis for developing a targeted cationic lipid delivery reagent for nucleic acids. Heme (ferric protoporphyrin IX) was conjugated to the aminolipid dioleoyl phosphatidylethanolamine (DOPE) and used to form cationic lipid particles with dioleoyl trimethylammonium propane (DOTAP). These lipids particles (DDH) protect oligoribonucleotides from degradation in human serum and increase oligoribonucleotide uptake into 2.2.15 human hepatoma cells (to a level of 50-60 ng oligo/10(4) cells) when compared with the same lipid particles (DD) prepared identically without heme. The DDH heme level that was optimal for oligoribonucleotide delivery was also optimal for maximum expression of plasmid-encoded luciferase. The enhancing effect of heme was evident only at net particle negative charge. Fluorescence microscopy showed that DDH delivered oligoribonucleotides into both the 2.2.15 cell cytoplasm and nucleus. DDH may thus be a potentially useful delivery vehicle for oligonucleotide-based therapeutics and transgenes, appropriate for use in such liver diseases as viral hepatitis, hepatoma, and hypercholesterolemia.
- Published
- 1997
- Full Text
- View/download PDF