Targeting of interstitial cells using a simple gene-transfer strategy.
BACKGROUND
Interstitial fibroblasts are central to the inflammatory response during the progression of tubulointerstitial fibrosis. We examined the efficiency of a new gene transfer method that targets interstitial cells by using parenchymal injection of DNA followed by electroporation.


METHODS
Fluoresceinisothiocyanate-labelled oligodeoxynucleotides (FITC-ODNs) or expression vectors were directly injected into the cortex of the kidney, followed by electroporation.


RESULTS
Transfection with FITC-ODNs or the EGFP expression vector resulted in efficient transfection in interstitial fibroblasts, but not in tubular epithelial cells or glomerular cells. Transfection efficiency was optimal after using a total of 150 microg of DNA in 1000 microl of PBS, combined with clamping of the renal vessels prior to electroporation. Gene expression peaked at 4 days after transfection and decreased by two orders of magnitude at 6 weeks post-transfection; however, expression recovered to near peak levels after parenchymal or intraperitoneal injection of FR901228, a histone deacetylase inhibitor.


CONCLUSIONS
We demonstrated that direct parenchymal injection of DNA combined with electroporation enables gene transfer into interstitial fibroblasts.
Fujii
Naohiko
N
Department of Advanced Technology for Transplantation, Osaka University Graduate School of Medicine, Suita 565-0871, Japan.
Isaka
Yoshitaka
Y
Takabatake
Yoshitsugu
Y
Mizui
Masayuki
M
Suzuki
Chigure
C
Takahara
Shiro
S
Ito
Takahito
T
Imai
Enyu
E
D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000008:administration & dosage / Q000494:pharmacology; D003239:Connective Tissue Cells / Q000187:drug effects / Q000378:metabolism; D047630:Depsipeptides / Q000008:administration & dosage / Q000494:pharmacology; D018274:Electroporation / Q000379:methods; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D016650:Fluorescein-5-isothiocyanate / Q000008:administration & dosage / Q000737:chemistry; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007668:Kidney / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D007678:Kidney Glomerulus / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008297:Male; D009838:Oligodeoxyribonucleotides / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors; D014162:Transfection / Q000379:methods
D000903:Antibiotics, Antineoplastic; D047630:Depsipeptides; D009838:Oligodeoxyribonucleotides; D049452:Green Fluorescent Proteins; C087123:romidepsin; D008156:Luciferases; D016650:Fluorescein-5-isothiocyanate