In vivo-targeted gene delivery using antibody-based nonviral vector.
Tissue-specific gene transfer remains one of the main challenges to deliver genes into designated and/or disseminated cells. We have previously shown successful gene transfer with a nonviral gene delivery system based on the simple chemical conjugation of plasmid DNA with antibody. However, this approach was hampered by low efficiency due to the poor translocation rate of DNA to the nucleus. To improve this approach, we have modified our vector by introducing noncovalent binding between the antibody and DNA, allowing the possibility to introduce different important molecules. The noncovalent association was achieved with neutravidin and biotinylated components: (1) biotinylated antibodies; (2) a biotinylated hemagglutinin fusogenic peptide of influenza virus to favor endosomal escape; and (3) biotinylated histone H1 to compact, protect, and associate DNA to the complex. We report here that this delivery system can be internalized by tumor cells targeted by a specific monoclonal antibody, permits the protection of the transfected DNA, and allows its subsequent transfer into the nucleus after escape from the endosomal compartment. We also demonstrate that, in vitro, gene transfer with this vector showed much higher reporter activity in cells (15 vs. 0.5%) and a stronger production of murine interleukin 2 as compared with our previous vector. In vivo, a single intravenous injection of the vector containing an antibody directed to the G250 renal cell carcinoma-associated antigen led to beta-galactosidase expression in engrafted tumor bearing G250 but not in G250-negative tumor or in other tissues. Altogether, these results indicate that our antibody-based vector is suitable to promote gene delivery in vitro and in vivo in tumor cells.
Déas
Olivier
O
INSERM U542/Paris-Sud University, Bâtiment Lavoisier, 16 avenue Paul Vaillant Couturier, 94807 Villejuif Cedex, France.
Angevin
Eric
E
Cherbonnier
Claire
C
Senik
Anna
A
Charpentier
Bernard
B
Levillain
Jean Paul
JP
Oosterwijk
Egbert
E
Hirsch
François
F
Dürrbach
Antoine
A
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism*; D019921:Biotinylation; D002467:Cell Nucleus / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D003850:Deoxyribonuclease I / Q000378:metabolism; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006389:Hemagglutinins, Viral / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D006801:Humans; D008164:Luminescent Proteins / Q000378:metabolism; D051379:Mice; D014407:Tumor Cells, Cultured; D001616:beta-Galactosidase / Q000378:metabolism
D000911:Antibodies, Monoclonal; C106533:G250 monoclonal antibody; D006389:Hemagglutinins, Viral; D006657:Histones; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D004247:DNA; D003850:Deoxyribonuclease I; D001616:beta-Galactosidase