Development of lipopolyplexes for gene delivery: a comparison of the effects of differing modes of targeting peptide display on the structure and transfection activities of lipopolyplexes

Bofinger, Robin; Zaw-Thin, May; Mitchell, Nicholas J.; Patrick, P. Stephen; Stowe, Cassandra; Gomez-Ramirez, Ana; Hailes, Helen C.; Kalber, Tammy L.; Tabor, Alethea B.

Abstract

The design, synthesis and formulation of non‐viral gene delivery vectors is an area of renewed research interest. Amongst the most efficient non‐viral gene delivery systems are lipopolyplexes, in which cationic peptides are co‐formulated with plasmid DNA and lipids. One advantage of lipopolyplex vectors is that they have the potential to be targeted to specific cell types by attaching peptide targeting ligands on the surface, thus increasing both the transfection efficiency and selectivity for disease targets such as cancer cells. In this paper, we have investigated two different modes of displaying cell‐specific peptide targeting ligands at the surface of lipopolyplexes. Lipopolyplexes formulated with bimodal peptides, with both receptor binding and DNA condensing sequences, were compared with lipopolyplexes with the peptide targeting ligand directly conjugated to one of the lipids. Three EGFR targeting peptide sequences were studied, together with a range of lipid formulations and maleimide lipid structures. The biophysical properties of the lipopolyplexes and their transfection efficiencies in a basal‐like breast cancer cell line were investigated using plasmid DNA bearing genes for the expression of firefly luciferase and green fluorescent protein. Fluorescence quenching experiments were also used to probe the macromolecular organisation of the peptide and pDNA components of the lipopolyplexes. We demonstrated that both approaches to lipopolyplex targeting give reasonable transfection efficiencies, and the transfection efficiency of each lipopolyplex formulation is highly dependent on the sequence of the targeting peptide. To achieve maximum therapeutic efficiency, different peptide targeting sequences and lipopolyplex architectures should be investigated for each target cell type.

Citation

Bofinger, R., Zaw-Thin, M., Mitchell, N. J., Patrick, P. S., Stowe, C., Gomez-Ramirez, A., …Tabor, A. B. (2018). Development of lipopolyplexes for gene delivery: a comparison of the effects of differing modes of targeting peptide display on the structure and transfection activities of lipopolyplexes. Journal of Peptide Science, e3131. https://doi.org/10.1002/psc.3131

• Journal Article Type: Article
• Acceptance Date: Sep 14, 2018
• Online Publication Date: Oct 16, 2018
• Publication Date: Oct 16, 2018
• Deposit Date: Oct 23, 2018
• Publicly Available Date: Oct 23, 2018
• Journal: Journal of Peptide Science
• Print ISSN: 1075-2617
• Publisher: Wiley
• Peer Reviewed: Peer Reviewed
• Pages: e3131
• DOI: https://doi.org/10.1002/psc.3131
• Keywords: Organic Chemistry; Molecular Medicine; Biochemistry; Molecular Biology; Pharmacology; Structural Biology; Drug Discovery; General Medicine
• Public URL: https://nottingham-repository.worktribe.com/output/1181626
• Publisher URL: https://onlinelibrary.wiley.com/doi/abs/10.1002/psc.3131
• Contract Date: Oct 23, 2018