title: The role of the helper lipid on the DNA transfection efficiency of lipopolyplex formulations.
abstract: Multifunctional, lipopolyplex formulations comprising a mixture of cationic liposomes and cationic, receptor-targeting peptides have potential use in gene therapy applications. Lipopolyplex formulations described here are typically far more efficient transfection agents than binary lipoplex or polyplex formulations. It has been shown previously that the peptide component mediates both DNA packaging and targeting of the nanoparticle while in this report we investigate the contribution of the lipid component. We hypothesised that the lipid components synergise with the peptides in the transfection process by promoting endosomal escape after lipid bilayer fusion. Lipopolyplexes were prepared with cationic liposomes comprising DOTAP with either neutral lipid DOPE or DOPC. DOPE promotes fusogenic, inverted hexagonal lipid structures while DOPC promotes more stable laminar structures. Lipopolyplexes containing DOPE showed substantially higher transfection efficiency than those formulated with DOPC, both in vitro and in vivo. DOPE-containing lipopolyplexes showed rapid endosomal trafficking and nuclear accumulation of DNA while DOPC-containing formulations remained within the late endo-lysosomal compartments. These findings are consistent with previous finding for the role of DOPE in lipoplexes and support the hypothesis regarding the function of the lipid components in lipopolyplexes. These findings will help to inform future lipopolyplex design, strategies and clinical development processes.
authors.affiliation: Experimental and Personalised Medicines Section, UCL Institute of Child Health, 30 Guilford Street, London WC1N 1EH, UK.
authors.forename: Zixiu
authors.initials: Z
authors.lastname: Du
authors.affiliation: Experimental and Personalised Medicines Section, UCL Institute of Child Health, 30 Guilford Street, London WC1N 1EH, UK.
authors.forename: Mustafa M
authors.initials: MM
authors.lastname: Munye
authors.affiliation: Experimental and Personalised Medicines Section, UCL Institute of Child Health, 30 Guilford Street, London WC1N 1EH, UK.
authors.forename: Aristides D
authors.initials: AD
authors.lastname: Tagalakis
authors.affiliation: Experimental and Personalised Medicines Section, UCL Institute of Child Health, 30 Guilford Street, London WC1N 1EH, UK.
authors.forename: Maria D I
authors.initials: MD
authors.lastname: Manunta
authors.affiliation: Experimental and Personalised Medicines Section, UCL Institute of Child Health, 30 Guilford Street, London WC1N 1EH, UK.
authors.forename: Stephen L
authors.initials: SL
authors.lastname: Hart
mesh_terms: D000818:Animals; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D004247:DNA / Q000737:chemistry; D011992:Endosomes / Q000378:metabolism; D005229:Fatty Acids, Monounsaturated / Q000737:chemistry; D005260:Female; D006801:Humans; D008051:Lipid Bilayers / Q000008:administration & dosage / Q000737:chemistry*; D008081:Liposomes / Q000008:administration & dosage / Q000737:chemistry*; D008168:Lung / Q000378:metabolism; D008561:Membrane Fusion; D051379:Mice; D010455:Peptides / Q000737:chemistry; D010713:Phosphatidylcholines / Q000737:chemistry*; D010714:Phosphatidylethanolamines / Q000737:chemistry*; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D020545:Respiratory Mucosa / Q000166:cytology / Q000378:metabolism; D014162:Transfection / Q000379:methods*
chemical_list: C410262:1,2-dioleoyl-glycero-3-phosphatidyl ethanolamine; D005229:Fatty Acids, Monounsaturated; D008051:Lipid Bilayers; D008081:Liposomes; D010455:Peptides; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds; D004247:DNA; C017251:1,2-oleoylphosphatidylcholine; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane