Cancer gene therapy using plasmid DNA: pharmacokinetic study of DNA following injection in mice.
The fate of plasmid DNA complexed with cationic lipids delivered intravenously in mice was evaluated at selected timepoints up to 6 months postinjection. Blood half-life and tissue distribution of plasmid DNA and potential expression in tissues were examined. Southern blot analyses of blood indicated that intact plasmid DNA was rapidly degraded, with a half-life of less than 5 min for intact plasmid, and was no longer detectable at 1 hr postinjection. Southern analyses of tissue demonstrated that intact DNA was differentially retained in the lung, spleen, liver, heart, kidney, marrow, and muscle up to 24 hr postinjection. After 7 days, no intact plasmid DNA was detectable by Southern blot analysis; however, the plasmid was detectable by the polymerase chain reaction (PCR) in all tissues examined at 7 and 28 days postinjection. At 6 months postinjection, femtogram levels of plasmid were detected only in muscle. Immunohistochemical analyses did not detect encoded protein in the tissues harboring residual plasmid at 1 or 7 days postinjection.
Lew
D
D
Vical Inc., San Diego, CA 92121, USA.
Parker
S E
SE
Latimer
T
T
Abai
A M
AM
Kuwahara-Rundell
A
A
Doh
S G
SG
Yang
Z Y
ZY
Laface
D
D
Gromkowski
S H
SH
Nabel
G J
GJ
D000818:Animals; D001483:Base Sequence; D002412:Cations / Q000737:chemistry; D002478:Cells, Cultured; D004247:DNA; D004274:DNA, Recombinant; D005260:Female; D005805:Genes, MHC Class I; D015316:Genetic Therapy; D015793:HLA-B7 Antigen / Q000235:genetics*; D006801:Humans; D007150:Immunohistochemistry; D007275:Injections, Intravenous; D008055:Lipids / Q000008:administration & dosage / Q000097:blood / Q000737:chemistry / Q000493:pharmacokinetics* / Q000494:pharmacology; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D008969:Molecular Sequence Data; D009369:Neoplasms / Q000628:therapy*; D010957:Plasmids / Q000008:administration & dosage / Q000097:blood / Q000493:pharmacokinetics*; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D014018:Tissue Distribution; D014162:Transfection; D014407:Tumor Cells, Cultured; D001613:beta 2-Microglobulin / Q000235:genetics
C096492:Allovectin-7; D002412:Cations; D004274:DNA, Recombinant; D015793:HLA-B7 Antigen; D008055:Lipids; D001613:beta 2-Microglobulin; D004247:DNA