DNA/polyethyleneimine/hyaluronic acid small complex particles and tumor suppression in mice.
The highest barriers for non-viral vectors to an efficient in vivo gene transfection would be (1) non-specific interaction with biological molecules, and (2) large size of the DNA complex particles. Protective coating of the DNA/polyethyleneimine (PEI) complexes by hyaluronic acid (HA) effectively diminished the adverse interactions with biological molecules. Here we found HA also protected the DNA/PEI complexes against aggregation and inactivation through lyophilization-and-rehydration procedures. It allows us to prepare the concentrated very small DNA complex particles (<70 nm) suspension by preparing the complexes at highly diluted conditions, followed by lyophilized-and-rehydrated to a small volume. In vivo gene expression efficiency of the small complex was examined with mice subcutaneously inoculated with B16 melanoma cells. These formulations showed high reporter-gene expression level in tumor after intravenous injection into tumor-bearing mice. Small complex was then made of the plasmid encoding GM-CSF gene, and injected into the mice bearing subcutaneous solid B16 tumor. After intravenous injection, it induced apparent tumor growth suppression in 50% of the mice. Notably, significant therapeutic effect was detected in the mice that received intratumoral injection, and 75% of the mice were completely cured with disappearance of tumor.
Ito
Tomoko
T
Department of Textile Science, Otsuma Women's University, Chiyoda-ku, Tokyo, Japan.
Yoshihara
Chieko
C
Hamada
Katsuyuki
K
Koyama
Yoshiyuki
Y
D000818:Animals; D045744:Cell Line, Tumor; D004247:DNA / Q000378:metabolism*; D005612:Freeze Drying; D005615:Freezing; D015870:Gene Expression / Q000187:drug effects; D020868:Gene Silencing / Q000187:drug effects; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics / Q000627:therapeutic use; D006820:Hyaluronic Acid / Q000494:pharmacology*; D008156:Luciferases / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence; D009369:Neoplasms / Q000473:pathology / Q000628:therapy*; D010957:Plasmids / Q000235:genetics; D011094:Polyethyleneimine / Q000494:pharmacology*; D014162:Transfection
D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D011094:Polyethyleneimine; D006820:Hyaluronic Acid; D004247:DNA; D008156:Luciferases