A novel gene therapy to the graft organ by a rapid injection of naked DNA I: long-lasting gene expression in a rat model of limb transplantation.
BACKGROUND
It is important to develop a nontoxic gene transfer method for immunosuppressed patients. In this study, the authors applied a nonviral gene transfer method using rapid injection of naked DNA into the graft limb in rats.


METHODS
Naked DNA (beta-galactosidase, luciferase, or green fluorescent protein expressing plasmid) was used to test an intravascular gene transfer approach in various conditions on the Lewis rat limb. Then, in a rat limb transplantation model, these marker genes were administered preoperatively (day -2) or perioperatively (day 0) to the graft limb by the authors' "venous protocol." The expression level of luciferase was observed over a long period using a noninvasive living image acquisition IVIS system.


RESULTS
Effective intravascular delivery of gene to the rat limb was achieved by a rapid bolus injection of naked DNA through the femoral caudal epigastric vein. Using this procedure, the limb graft with the marker gene perioperatively in place was safely transplanted. After limb transplantation, sustained marker gene expression was observed for more than 2 months.


CONCLUSIONS
This is the first report showing that the method of rapid injection of naked DNA into the limb can be applied to gene modification for organ transplantation.
Sato
Yuki
Y
Division of Organ Replacement Research, Center for Molecular Medicine, Jichi Medical School, Yakushiji, Minamikawachi, Tochigi, Japan.
Ajiki
Takashi
T
Inoue
Seiichiro
S
Hakamata
Yoji
Y
Murakami
Takashi
T
Kaneko
Takashi
T
Takahashi
Masafumi
M
Kobayashi
Eiji
E
D000818:Animals; D004247:DNA / Q000008:administration & dosage / Q000235:genetics*; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D006085:Graft Survival / Q000502:physiology*; D049452:Green Fluorescent Proteins; D006614:Hindlimb / Q000637:transplantation*; D007275:Injections, Intravenous; D008164:Luminescent Proteins / Q000235:genetics; D051381:Rats; D011917:Rats, Inbred Lew; D016038:Skin Transplantation; D013997:Time Factors; D014185:Transplantation, Isogeneic / Q000502:physiology*; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D004247:DNA; D001616:beta-Galactosidase