title: Improved intracellular delivery of peptide- and lipid-nanoplexes by natural glycosides.
abstract: Targeted nanocarriers undergo endocytosis upon binding to their membrane receptors and are transported into cellular compartments such as late endosomes and lysosomes. In gene delivery the genetic material has to escape from the cellular compartments into the cytosol. The process of endosomal escape is one of the most critical steps for successful gene delivery. For this reason synthetic lipids with fusogenic properties such as 2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) are integrated into the nanocarriers. In this study we show that a natural, plant derived glycoside (SO1861) from Saponaria officinalis L. greatly improves the efficacy of lipid based as well as non-lipid based targeted nanoplexes consisting of a targeted K16 peptide with a nucleic acid binding domain and plasmid-DNA, minicircle-DNA or small interfering RNA (siRNA). By confocal live cell imaging and single cell analyses, we demonstrate that SO1861 augments the escape of the genetic cargo out of the intracellular compartments into the cytosol. Co-localisation experiments with fluorescence labelled dextran and transferrin indicate that SO1861 induces the release of the genetic cargo out of endosomes and lysosomes. However, the transduction efficacy of a lentivirus based gene delivery system was not augmented. In order to design receptor-targeted nanoplexes (LPD) with improved functional properties, SO1861 was integrated into the lipid matrix of the LPD. The SO1861 sensitized LPD (LPDS) were characterized by dynamic light scattering and transmission electron microscopy. Compared to their LPD counterparts the LPDS-nanoplexes showed a greatly improved gene delivery. As shown by differential scanning calorimetry SO1861 can be easily integrated into the lipid bilayer of glycerophospholipid model membranes. This underlines the great potential of SO1861 as a new transfection multiplier for non-viral gene delivery systems.
authors.affiliation: Wolfson Centre for Gene Therapy of Childhood Disease, UCL Institute of Child Health, University College London, 30 Guilford Street, London WC1N 1EH, United Kingdom; Institute of Pharmacy, Free University Berlin, Königin-Luise-Str. 2+4, D-14195 Berlin, Germany. Electronic address: weng@zedat.fu-berlin.de.
authors.forename: Alexander
authors.initials: A
authors.lastname: Weng
authors.affiliation: Wolfson Centre for Gene Therapy of Childhood Disease, UCL Institute of Child Health, University College London, 30 Guilford Street, London WC1N 1EH, United Kingdom.
authors.forename: Maria D I
authors.initials: MD
authors.lastname: Manunta
authors.affiliation: Institut für Laboratoriumsmedizin, Klinische Chemie und Pathobiochemie, Charité - Universitätsmedizin Berlin, Augustenburger Platz 1, D-13353 Berlin, Germany.
authors.forename: Mayank
authors.initials: M
authors.lastname: Thakur
authors.affiliation: Institute of Pharmacy, Free University Berlin, Königin-Luise-Str. 2+4, D-14195 Berlin, Germany.
authors.forename: Roger
authors.initials: R
authors.lastname: Gilabert-Oriol
authors.affiliation: Wolfson Centre for Gene Therapy of Childhood Disease, UCL Institute of Child Health, University College London, 30 Guilford Street, London WC1N 1EH, United Kingdom.
authors.forename: Aristides D
authors.initials: AD
authors.lastname: Tagalakis
authors.affiliation: Wolfson Centre for Gene Therapy of Childhood Disease, UCL Institute of Child Health, University College London, 30 Guilford Street, London WC1N 1EH, United Kingdom.
authors.forename: Ayad
authors.initials: A
authors.lastname: Eddaoudi
authors.affiliation: Wolfson Centre for Gene Therapy of Childhood Disease, UCL Institute of Child Health, University College London, 30 Guilford Street, London WC1N 1EH, United Kingdom.
authors.forename: Mustafa M
authors.initials: MM
authors.lastname: Munye
authors.affiliation: Wolfson Centre for Gene Therapy of Childhood Disease, UCL Institute of Child Health, University College London, 30 Guilford Street, London WC1N 1EH, United Kingdom.
authors.forename: Conrad A
authors.initials: CA
authors.lastname: Vink
authors.affiliation: Leibniz-Institut für Molekulare Pharmakologie (FMP), Robert-Rössle-Str. 10, 13125 Berlin, Germany.
authors.forename: Burkhard
authors.initials: B
authors.lastname: Wiesner
authors.affiliation: Leibniz-Institut für Molekulare Pharmakologie (FMP), Robert-Rössle-Str. 10, 13125 Berlin, Germany.
authors.forename: Jenny
authors.initials: J
authors.lastname: Eichhorst
authors.affiliation: Institute of Pharmacy, Free University Berlin, Königin-Luise-Str. 2+4, D-14195 Berlin, Germany.
authors.forename: Matthias F
authors.initials: MF
authors.lastname: Melzig
authors.affiliation: Wolfson Centre for Gene Therapy of Childhood Disease, UCL Institute of Child Health, University College London, 30 Guilford Street, London WC1N 1EH, United Kingdom. Electronic address: s.hart@ucl.ac.uk.
authors.forename: Stephen L
authors.initials: SL
authors.lastname: Hart
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D004247:DNA / Q000008:administration & dosage*; D011992:Endosomes / Q000378:metabolism; D018014:Gene Transfer Techniques; D006027:Glycosides / Q000737:chemistry*; D006801:Humans; D008055:Lipids / Q000737:chemistry*; D051379:Mice; D008969:Molecular Sequence Data; D053758:Nanoparticles / Q000737:chemistry; D010455:Peptides / Q000737:chemistry*; D010957:Plasmids / Q000008:administration & dosage*; D034741:RNA, Small Interfering / Q000008:administration & dosage*; D029750:Saponaria / Q000737:chemistry; D014162:Transfection
chemical_list: D006027:Glycosides; D008055:Lipids; D010455:Peptides; D034741:RNA, Small Interfering; D004247:DNA
keywords: Endosomal escape; Gene delivery; Natural glycoside