Efficient gene expression in skin wound sites following local plasmid injection.
Transfection of the skin by local gene delivery, as well as widespread transfection of systemic tissues following intravenous injection of cationic liposome/DNA complexes have been reported. Here, we show that surgically wounded mouse skin can be transfected either by local injection of DNA alone or by intravenous injection of optimized cationic liposome/DNA complexes; however, direct cutaneous injection produces much higher levels of gene expression in the skin, which is targeted to dermal and subdermal layers. High levels of chloramphenicol acetyltransferase activity were present from 3 h to 2 wk following direct injection of a gene expression plasmid into wounded skin and were maintained at detectable levels up to 8 wk after injection. Expression of transferred chloramphenicol acetyltransferase as well as beta-GAL genes was localized to fibroblasts, macrophages, and adipocytes as determined by histochemistry and immunohistochemistry. Further- more, local injection of a human granulocyte- colony-stimulating factor gene expression plasmid produced high levels of the biologically relevant human granulocyte-colony-stimulating factor protein in wounded mouse skin. This efficient and simple method of site-specific gene transfer into wounds may lead to the development of cutaneous gene therapy directed against disorders of abnormal cutaneous wound healing.
Meuli
M
M
Department of Surgery, University Children's Hospital Zurich, Zurich, Switzerland.
Liu
Y
Y
Liggitt
D
D
Kashani-Sabet
M
M
Knauer
S
S
Meuli-Simmen
C
C
Harrison
M R
MR
Adzick
N S
NS
Heath
T D
TD
Debs
R J
RJ
D000818:Animals; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D003587:Cytomegalovirus / Q000235:genetics; D004279:DNA, Viral / Q000032:analysis; D004305:Dose-Response Relationship, Drug; D005260:Female; D015870:Gene Expression; D016179:Granulocyte Colony-Stimulating Factor / Q000235:genetics; D007267:Injections; D051379:Mice; D008813:Mice, Inbred ICR; D010957:Plasmids / Q000008:administration & dosage*; D013997:Time Factors; D014162:Transfection; D014945:Wound Healing / Q000235:genetics*; D014947:Wounds and Injuries / Q000235:genetics*
D004279:DNA, Viral; D016179:Granulocyte Colony-Stimulating Factor; D015500:Chloramphenicol O-Acetyltransferase