Nonviral in vivo gene delivery into tumors using a novel low volume jet-injection technology.
The jet-injection technology has developed as an applicable alternative to viral or liposomal gene delivery systems. In this study a novel, low-volume, 'high-speed jet injector' hand-held system was used for the direct gene transfer of naked DNA into tumors. Lewis-lung carcinoma bearing mice were jet-injected with the beta-galactosidase (LacZ), the green fluorescence (GFP) or the human tumor necrosis factor alpha (TNF-alpha) gene carrying vector plasmids. The animals received five jet injections into the tumor at a pressure of 3.0 bar, delivering 3--5 microl plasmid DNA (1 microg DNA/microl in water) per single jet injection. The jet injection of DNA leads to a widespread expression pattern within tumor tissues with penetration depths of 5--10 mm. Analysis of tumor cryosections revealed moderate LacZ or GFP expression at 48 h and strong reporter gene expression 72 h and 96 h after jet injection. The simultaneous jet injection of the TNF-alpha and LacZ carrying vectors demonstrated efficient expression and secretion of both the cytokine, as well as LacZ expression within the tumor 24 h, 48 h, 72 h, 96 h and 120 h after jet injection. These studies demonstrate the applicability of jet injection for the efficient in vivo gene transfer into tumors for nonviral gene therapy of cancer using minimal amounts of naked DNA.
Walther
W
W
Max-Delbrück-Center for Molecular Medicine, Berlin, Germany.
Stein
U
U
Fichtner
I
I
Malcherek
L
L
Lemm
M
M
Schlag
P M
PM
D000818:Animals; D018827:Carcinoma, Lewis Lung / Q000378:metabolism / Q000628:therapy; D005260:Female; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D049452:Green Fluorescent Proteins; D006801:Humans; D007202:Indicators and Reagents; D007277:Injections, Jet; D007763:Lac Operon / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D008815:Mice, Inbred Strains; D009368:Neoplasm Transplantation; D009369:Neoplasms / Q000628:therapy*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism
D007202:Indicators and Reagents; D008164:Luminescent Proteins; D014409:Tumor Necrosis Factor-alpha; D049452:Green Fluorescent Proteins