Stabilized plasmid-lipid particles for systemic gene therapy.
The structure of 'stabilized plasmid-lipid particles' (SPLP) and their properties as systemic gene therapy vectors has been investigated. We show that SPLP can be visualized employing cryo-electron microscopy to be homogeneous particles of diameter 72 +/- 5 nm consisting of a lipid bilayer surrounding a core of plasmid DNA. It is also shown that SPLP exhibit long circulation lifetimes (circulation half-life >6 h) following intravenous (i.v.) injection in a murine tumor model resulting in accumulation of up to 3% of the total injected dose and concomitant reporter gene expression at a distal (hind flank) tumor site. In contrast, i v. injection of naked plasmid DNA or plasmid DNA-cationic liposome complexes did not result in significant plasmid delivery to the tumor site or gene expression at that site. Furthermore, it is shown that high doses of SPLP corresponding to 175 microg plasmid per mouse are nontoxic as assayed by monitoring serum enzyme levels, whereas i.v. injection of complexes give rise to significant toxicity at dose levels above 20 microg plasmid per mouse. It is concluded that SPLP exhibit properties consistent with potential utility as a nontoxic systemic gene therapy vector.
Tam
P
P
Inex Pharmaceuticals Corporation, Burnaby, BC, Canada.
Monck
M
M
Lee
D
D
Ludkovski
O
O
Leng
E C
EC
Clow
K
K
Stark
H
H
Scherrer
P
P
Graham
R W
RW
Cullis
P R
PR
D000818:Animals; D020285:Cryoelectron Microscopy; D005260:Female; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D007275:Injections, Intravenous; D008055:Lipids; D008156:Luciferases / Q000235:genetics; D008175:Lung Neoplasms / Q000628:therapy*; D051379:Mice; D008810:Mice, Inbred C57BL; D009374:Neoplasms, Experimental / Q000628:therapy*; D010957:Plasmids
D008055:Lipids; D008156:Luciferases