The role of ex-vivo gene therapy of vein grafts with Egr-1 decoy in the suppression of intimal hyperplasia.
OBJECTIVE
To test the hypothesis that vein graft intimal hyperplasia can be significantly suppressed by a single intra-operative transfection of the graft with a decoy oligonucleotide (ODN) binding the transcription factor Egr-1.


METHODS
Experimental study.


METHODS
Jugular vein to carotid artery interposition grafts in rabbits were treated with Egr-1 decoy, mutant decoy ODN, vehicle alone, using a non-distending pressure of 300 mm Hg for 20 min, or were left untreated. All animals were fed a 2% cholesterol diet. The animals were sacrificed after 48h, 6 weeks and 12 weeks. Paraffin-embedded vein sections were subjected to angiometric analysis.


RESULTS
Successful delivery of the ODN was confirmed by DAPI staining. Quantitative real-time PCR revealed a 60% decrease of the Egr-1 gene expression in the animals in which the Egr-1 decoy ODN was delivered. Cellular proliferation was also significantly decreased as indicated by the Ki-67 labelling index. An increase in intimal and medial thickness was found in all vein grafts. However, intimal thickness was significantly reduced in the grafts treated with Egr-1 decoy ODN, whereas luminal area was significantly increased.


CONCLUSIONS
A single intra-operative pressure-mediated transfection of vein grafts with Egr-1 decoy ODN significantly suppresses intimal hyperplasia in a rabbit hypercholesterolaemic model.
Peroulis
M
M
Center of Experimental Surgery, Foundation of Biomedical Research, Academy of Athens, Athens, Greece.
Kakisis
J
J
Kapelouzou
A
A
Giagini
A
A
Giaglis
S
S
Mantziaras
G
G
Kostomitsopoulos
N
N
Karayannacos
P
P
Macheras
A
A
D000818:Animals; D002339:Carotid Arteries / Q000601:surgery; D049109:Cell Proliferation; D004195:Disease Models, Animal; D051766:Early Growth Response Protein 1 / Q000235:genetics*; D005786:Gene Expression Regulation; D015316:Genetic Therapy / Q000379:methods*; D006083:Graft Occlusion, Vascular / Q000235:genetics / Q000517:prevention & control*; D006965:Hyperplasia; D007432:Intraoperative Period; D007601:Jugular Veins / Q000601:surgery; D008297:Male; D009841:Oligonucleotides / Q000737:chemistry; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D017539:Tunica Intima / Q000473:pathology*
D051766:Early Growth Response Protein 1; D009841:Oligonucleotides