Therapeutic protein transduction of mammalian cells and mice by nucleic acid-free lentiviral nanoparticles.
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.
Link
Nils
N
Institute for Chemical and Bio-Engineering (ICB), Swiss Federal Institute of Technology, ETH Hoenggerberg, HCI F115, Wolfgang-Pauli-Strasse 10, CH-8093 Zurich, Switzerland.
Aubel
Corinne
C
Kelm
Jens M
JM
Marty
René R
RR
Greber
David
D
Djonov
Valentin
V
Bourhis
Jean
J
Weber
Wilfried
W
Fussenegger
Martin
M
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
D000970:Antineoplastic Agents; D009570:Nitriles; D011355:Prodrugs; D011993:Recombinant Fusion Proteins; C021514:cyanogenic beta-glucosidase; D001617:beta-Glucosidase; C005091:linamarin