title: Therapeutic protein transduction of mammalian cells and mice by nucleic acid-free lentiviral nanoparticles.
abstract: The straightforward production and dose-controlled administration of protein therapeutics remain major challenges for the biopharmaceutical manufacturing and gene therapy communities. Transgenes linked to HIV-1-derived vpr and pol-based protease cleavage (PC) sequences were co-produced as chimeric fusion proteins in a lentivirus production setting, encapsidated and processed to fusion peptide-free native protein in pseudotyped lentivirions for intracellular delivery and therapeutic action in target cells. Devoid of viral genome sequences, protein-transducing nanoparticles (PTNs) enabled transient and dose-dependent delivery of therapeutic proteins at functional quantities into a variety of mammalian cells in the absence of host chromosome modifications. PTNs delivering Manihot esculenta linamarase into rodent or human, tumor cell lines and spheroids mediated hydrolysis of the innocuous natural prodrug linamarin to cyanide and resulted in efficient cell killing. Following linamarin injection into nude mice, linamarase-transducing nanoparticles impacted solid tumor development through the bystander effect of cyanide.
authors.affiliation: Institute for Chemical and Bio-Engineering (ICB), Swiss Federal Institute of Technology, ETH Hoenggerberg, HCI F115, Wolfgang-Pauli-Strasse 10, CH-8093 Zurich, Switzerland.
authors.forename: Nils
authors.initials: N
authors.lastname: Link
authors.forename: Corinne
authors.initials: C
authors.lastname: Aubel
authors.forename: Jens M
authors.initials: JM
authors.lastname: Kelm
authors.forename: René R
authors.initials: RR
authors.lastname: Marty
authors.forename: David
authors.initials: D
authors.lastname: Greber
authors.forename: Valentin
authors.initials: V
authors.lastname: Djonov
authors.forename: Jean
authors.initials: J
authors.lastname: Bourhis
authors.forename: Wilfried
authors.initials: W
authors.lastname: Weber
authors.forename: Martin
authors.initials: M
authors.lastname: Fussenegger
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000378:metabolism; D002460:Cell Line; D045744:Cell Line, Tumor; D003131:Combined Modality Therapy; D006224:Cricetinae; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D008819:Mice, Nude; D049329:Nanostructures; D009369:Neoplasms / Q000188:drug therapy / Q000628:therapy*; D009570:Nitriles / Q000627:therapeutic use; D011355:Prodrugs / Q000627:therapeutic use; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D018874:Spheroids, Cellular; D014162:Transfection; D001617:beta-Glucosidase / Q000235:genetics* / Q000378:metabolism
chemical_list: D000970:Antineoplastic Agents; D009570:Nitriles; D011355:Prodrugs; D011993:Recombinant Fusion Proteins; C021514:cyanogenic beta-glucosidase; D001617:beta-Glucosidase; C005091:linamarin