title: Fast therapeutic DNA internalization - A high potential transfection system based on a peptide mimicking cationic lipid.
abstract: The delivery of nucleic acids into cells is a determining factor for successful gene therapy. In this study we investigate the uptake and time dependent processing of a lipid-based non-viral nucleic acid delivery system composed of a peptide-mimicking cationic lipid (N-{6-amino-1-[N-(9Z)-octadec-9-enylamino]-1-oxohexan-(2S)-2-yl}-N'-{2-[N,N-bis(2-aminoethyl)amino]ethyl}-2-hexadecylpropandiamide - OH4) and a phospholipid (1,2-dioleoyl-sn-glycero-3-phosphoethanolamine - DOPE). Studies by confocal laser scanning microscopy (CLSM) indicate a rapid internalization of fluorescent labelled DNA within 1h. Furthermore, vesicular structures on the lipid surface were reported, which are associated with the application of the lipid-based non-viral vector. Time dependent investigations of the gene expression of a reporter gene encoding for enhanced green fluorescent protein (eGFP) or luciferase in 4 different cell lines demonstrate an initial gene expression soon after 4h followed by a boost in gene expression beginning from 12h to 24h. Investigations with selective blocking of endocytic pathways using low molecular weight inhibitors suggested clathrin-mediated endocytosis as main internalization route in 3 cell lines. Our research presents a new horizon in rapid gene therapy using non-viral vectors; due to the modifications of the lipid components, fast nucleic acid internalizations could be achieved using our delivery systems.
authors.affiliation: Department of Pharmaceutics and Biopharmaceutics, University Marburg, Ketzerbach 63, 35037 Marburg, Germany; Martin Luther University Halle-Wittenberg, Institute of Pharmacy, Wolfgang-Langenbeck-Strasse 4, 06120 Halle, Saale, Germany.
authors.forename: Christopher
authors.initials: C
authors.lastname: Janich
authors.affiliation: Department of Pharmaceutics and Biopharmaceutics, University Marburg, Ketzerbach 63, 35037 Marburg, Germany.
authors.forename: Shashank Reddy
authors.initials: SR
authors.lastname: Pinnapireddy
authors.affiliation: Martin Luther University Halle-Wittenberg, Institute of Pharmacy, Wolfgang-Langenbeck-Strasse 4, 06120 Halle, Saale, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Erdmann
authors.affiliation: Institute of Pharmacy, Biomedical Materials Group, Martin Luther University Halle-Wittenberg, Heinrich-Damerow-Str. 4, 06120 Halle, Saale, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Groth
authors.affiliation: Martin Luther University Halle-Wittenberg, Institute of Pharmacy, Wolfgang-Langenbeck-Strasse 4, 06120 Halle, Saale, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Langner
authors.affiliation: Department of Pharmaceutics and Biopharmaceutics, University Marburg, Ketzerbach 63, 35037 Marburg, Germany.
authors.forename: Udo
authors.initials: U
authors.lastname: Bakowsky
authors.affiliation: Martin Luther University Halle-Wittenberg, Institute of Pharmacy, Wolfgang-Langenbeck-Strasse 4, 06120 Halle, Saale, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Wölk
mesh_terms: D000818:Animals; D002412:Cations / Q000737:chemistry; D045744:Cell Line, Tumor; D022163:Clathrin-Coated Vesicles / Q000378:metabolism; D004247:DNA / Q000008:administration & dosage*; D016503:Drug Delivery Systems / Q000379:methods*; D004705:Endocytosis / Q000187:drug effects*; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D008081:Liposomes; D008156:Luciferases / Q000378:metabolism; D018613:Microscopy, Confocal; D010316:Particle Size; D010714:Phosphatidylethanolamines / Q000737:chemistry; D034421:Sus scrofa; D014162:Transfection / Q000379:methods*
chemical_list: D002412:Cations; D008081:Liposomes; D010714:Phosphatidylethanolamines; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; C020888:1,2-dielaidoylphosphatidylethanolamine; D004247:DNA; D008156:Luciferases
keywords: Cationic liposomes; Endocytosis; Fast uptake; Membrane fusion; Nucleic acid delivery; Vesicles