Lipopolyplex ternary delivery systems incorporating C14 glycerol-based lipids
| Autor: | Alethea B. Tabor, Frederick Campbell, M. Jayne Lawrence, Barbara Fridrich, M. Firouz Mohd Mustapa, Katharina Welser, Tony Ng, Jimmy Ho, Helen C. Hailes, Laila Kudsiova, David J. Barlow, Melanie Keppler |
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| Rok vydání: | 2011 |
| Předmět: |
Chemical Phenomena
Membrane Fluidity Molecular Conformation Pharmaceutical Science Peptide Breast Neoplasms Glyceryl Ethers Endosomes Biology Ligands chemistry.chemical_compound Cell Line Tumor Drug Discovery Membrane fluidity Humans Particle Size Lipid bilayer Peptide sequence Fluorescent Dyes chemistry.chemical_classification Gel electrophoresis Cell Nucleus Gene Transfer Techniques Lipid metabolism Stereoisomerism Transfection DNA Lipid Metabolism Lipids Neoplasm Proteins chemistry Biochemistry Molecular Medicine lipids (amino acids peptides and proteins) Female Peptides Integrin alpha5beta1 Plasmids |
| Zdroj: | Molecular pharmaceutics. 8(5) |
| ISSN: | 1543-8392 |
| Popis: | The structure, biophysical properties and biological behavior of lipopolyplex ternary gene delivery vectors incorporating novel C14 glycerol based lipids of varying alkyl chain geometry (containing cis, trans or alkyne double bonds) have been studied in the presence and absence of a bifunctional targeting peptide designed to both condense DNA and confer integrin-specific targeting. In vitro transfection studies in breast cancer MDA-MB-231 cells revealed that ternary formulations of lipid:peptide:DNA (LPD) complexes prepared using the aforementioned lipids possessed highly synergistic transfection activity up to 2500-fold higher than their respective lipid:DNA (LD) or peptide:DNA (PD) counterparts. Furthermore, the small structural differences in the lipid alkyl chain geometries also resulted in pronounced differences in transfection within each type of formulation, whereby the trans lipids showed best activity when formulated as LD complexes, whereas the cis lipids were superior in LPD formulations. Confocal fluorescence internalization studies using labeled components of the formulations showed both the lipid and the DNA of LD complexes to be trapped in endocytic compartments, whereas in the case of LPD complexes, the DNA was clearly released from the endosomal compartments and, together with the peptide, internalized within the cell nucleus. Physicochemical characterization of the formulations carried out by light and neutron scattering, zeta potential measurement, and negative staining electron microscopy detected major structural differences between LD and LPD complexes. Gel electrophoresis assays additionally showed differences between the individual lipids tested in each type of formulation. In conclusion, the superior transfection of the trans lipids in the LD complexes was thought to be attributed to superior DNA binding caused by a more closely matched charge distribution of the more rigid, trans lipids with the DNA. In the case of the LPD complexes, the DNA was thought to be predominantly condensed by the cationic portion of the peptide forming a central core surrounded by a lipid bilayer from which the targeting sequence partially protrudes. The more fluid, cis lipids were thought to confer better activity in this formulation due to allowing more of the targeting peptide sequence to protrude. |
| Databáze: | OpenAIRE |
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