Polyion complex micelles as vectors in gene therapy--pharmacokinetics and in vivo gene transfer.
To establish non-viral gene delivery systems for intravenous administration, complexes of DNA and block copolymer consisting of poly-L-lysine and poly(ethylene glycol) were tested in in vivo turnover studies. The polyion complex micelles have self-assembling core-shell structures, yielding spherical nano-particles with small absolute values of zeta-potential. Southern blot analysis showed that supercoiled DNA was observed for 30 min and open circular or linear DNA was seen for 3 h after intravenous administration of PIC micelles having the charge ratios of 1:4 and PLL length of 48 mer. The PIC micelles with shorter PLL length showed lower stability in the blood stream suggesting that DNA is able to persist as an intact molecule in the blood stream using this system. Though having no ligands, PIC micelles with charge ratios of 1:2 and 1:4 transfected efficiently into HepG2 cells. Preincubation with free copolymer inhibited expression of the reporter gene, suggesting that adsorption of block copolymer to the cell surface blocked the interaction site of the PIC micelles. When the PIC micelles were injected via supramesenteric vein, expression of the gene was observed only in the liver and was sustained for 3 days. It was suggested that this gene delivery system is intrinsically efficient.
Harada-Shiba
M
M
National Cardiovascular Center Research Institute, Osaka, Japan.
Yamauchi
K
K
Harada
A
A
Takamisawa
I
I
Shimokado
K
K
Kataoka
K
K
D006528:Carcinoma, Hepatocellular / Q000628:therapy*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000493:pharmacokinetics*; D006801:Humans; D007275:Injections, Intravenous; D008113:Liver Neoplasms / Q000628:therapy*; D008156:Luciferases / Q000235:genetics; D008823:Micelles; D008854:Microscopy, Electron; D010957:Plasmids; D011092:Polyethylene Glycols; D011107:Polylysine; D013997:Time Factors; D014162:Transfection / Q000379:methods*; D014407:Tumor Cells, Cultured
D008823:Micelles; D011107:Polylysine; D011092:Polyethylene Glycols; D008156:Luciferases