title: Modifying plasmid-loaded HSA-nanoparticles with cell penetrating peptides - Cellular uptake and enhanced gene delivery.
abstract: Gene therapy bears great potential for the cure of a multitude of human diseases. Research efforts focussed on the use of viral delivery vectors in the past decades, neglecting non-viral gene therapies of physical or chemical origin due to low transfection efficiency. However, side effects such as activation of oncogenes and inflammatory reactions upon immune cell activation are major obstacles impeding the clinical applicability of viral gene therapy vectors. The aim of this study was the development of a non-viral gene delivery system based on plasmid-loaded human serum albumin nanoparticles, which are biocompatible, biodegradable, and non-toxic in relevant concentrations. The surface of said nanoparticles was modified with different cell penetrating peptides, namely Tat, nona-arginine R9, and the penetratin analogue EB1. We hypothesise that the surface modified nanoparticles can effectively enter HEK 293T cells based on the cell penetrating properties of the different peptides attached. A variety of inhibitors were used targeting distinct uptake pathways in an effort to understand the mechanisms utilized by the various cell penetrating peptides on the surface of the nanoparticles. A significant increase in transfection efficiency compared to free DNA or polyplexes was seen for these novel delivery vectors.
authors.affiliation: Institute of Pharmaceutical Technology and Biopharmacy, University of Münster, Corrensstraße 48, 48149 Münster, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Mesken
authors.affiliation: Institute of Pharmaceutical Science, King's College London, Franklin-Wilkins Building, 150 Stamford Street, London, SE1 9NH, UK.
authors.forename: A
authors.initials: A
authors.lastname: Iltzsche
authors.affiliation: Institute of Pharmaceutical Technology and Biopharmacy, University of Münster, Corrensstraße 48, 48149 Münster, Germany.
authors.forename: D
authors.initials: D
authors.lastname: Mulac
authors.affiliation: Institute of Pharmaceutical Technology and Biopharmacy, University of Münster, Corrensstraße 48, 48149 Münster, Germany. Electronic address: k.langer@wwu.de.
authors.forename: K
authors.initials: K
authors.lastname: Langer
mesh_terms: D002470:Cell Survival / Q000187:drug effects; D057846:Cell-Penetrating Peptides / Q000008:administration & dosage* / Q000737:chemistry*; D004247:DNA / Q000008:administration & dosage / Q000737:chemistry; D005079:Excipients; D015696:Gene Products, tat / Q000737:chemistry; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D057809:HEK293 Cells; D006801:Humans; D053758:Nanoparticles / Q000737:chemistry*; D010957:Plasmids / Q000737:chemistry*; D012709:Serum Albumin / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000633:toxicity; D014162:Transfection
chemical_list: D057846:Cell-Penetrating Peptides; D005079:Excipients; D015696:Gene Products, tat; D012709:Serum Albumin; D004247:DNA
keywords: Cell penetrating peptides; Cell transfection; Cellular uptake; Human serum albumin (HSA); Plasmid DNA; Transfection