Long-term stabilization of vein graft wall architecture and prolonged resistance to experimental atherosclerosis after E2F decoy oligonucleotide gene therapy.
OBJECTIVE
We tested the hypothesis that a single intraoperative transfection of rabbit vein grafts with a decoy oligonucleotide that blocks cell-cycle gene transactivation by the transcription factor E2F induces long-term stable adaptation that involves medial hypertrophy and a resistance to neointimal hyperplasia and atherosclerosis.


METHODS
Jugular vein to carotid artery interposition vein grafts in hypercholesterolemic rabbits were treated, using pressure-mediated delivery, with either E2F decoy oligonucleotide, scrambled oligonucleotide, or vehicle alone. E2F decoy inhibition of cell-cycle gene expression was determined by measuring proliferating cell nuclear antigen upregulation and bromodeoxyuridine incorporation in vascular smooth muscle cells. Neointimal hyperplasia and atherosclerosis were compared between groups at 6 months after operation. Wall stress was derived from the ratio of luminal radius to wall thickness. Normal rabbits exposed to 6 weeks of diet-induced hypercholesterolemia starting 6 months after operation were analyzed in the same manner.


RESULTS
The E2F decoy oligonucleotide, but not scrambled oligonucleotide or vehicle alone, inhibited proliferating cell nuclear antigen expression and smooth muscle cell proliferation. Furthermore, this manipulation of cell-cycle gene expression yielded an inhibition of neointimal hyperplasia and atherosclerotic plaque formation throughout the 6 months of cholesterol feeding. In normocholesterolemic rabbits, vehicle-treated and scrambled oligonucleotide-treated vein grafts remain susceptible to diet-induced atherosclerosis as well, whereas resistance to this disease induction remained stable in genetically engineered grafts.


CONCLUSIONS
A single intraoperative pressure-mediated delivery of E2F decoy effectively provides vein grafts with long-term resistance to neointimal hyperplasia and atherosclerosis. These findings suggest that long-term reduction in human vein graft failure rates may be feasible with this ex vivo gene therapy approach.
Ehsan
A
A
Division of Cardiovascular Medicine, Brigham and Women's Hospital/Harvard Medical School, 75 Francis Street, Boston, MA 202115, USA.
Mann
M J
MJ
Dell'Acqua
G
G
Dzau
V J
VJ
D000714:Anastomosis, Surgical; D000818:Animals; D000963:Antimetabolites / Q000493:pharmacokinetics / Q000627:therapeutic use; D001161:Arteriosclerosis / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism / Q000628:therapy*; D001973:Bromodeoxyuridine / Q000493:pharmacokinetics / Q000627:therapeutic use; D002340:Carotid Artery Diseases / Q000139:chemically induced / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D017536:Carotid Artery, Common / Q000601:surgery*; D002352:Carrier Proteins; D018797:Cell Cycle Proteins / Q000235:genetics / Q000627:therapeutic use*; D002455:Cell Division / Q000187:drug effects; D002791:Cholesterol, Dietary / Q000633:toxicity; D015342:DNA Probes / Q000737:chemistry; D004268:DNA-Binding Proteins; D018450:Disease Progression; D050684:E2F Transcription Factors; D015316:Genetic Therapy / Q000379:methods*; D006984:Hypertrophy; D007601:Jugular Veins / Q000378:metabolism / Q000473:pathology / Q000637:transplantation*; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000473:pathology; D018809:Proliferating Cell Nuclear Antigen / Q000187:drug effects / Q000378:metabolism; D011817:Rabbits; D056465:Retinoblastoma-Binding Protein 1; D050680:Transcription Factor DP1; D014157:Transcription Factors / Q000235:genetics / Q000627:therapeutic use*; D014162:Transfection; D017539:Tunica Intima / Q000187:drug effects / Q000473:pathology; D015854:Up-Regulation / Q000187:drug effects
D000963:Antimetabolites; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D002791:Cholesterol, Dietary; D015342:DNA Probes; D004268:DNA-Binding Proteins; D050684:E2F Transcription Factors; D018809:Proliferating Cell Nuclear Antigen; D056465:Retinoblastoma-Binding Protein 1; D050680:Transcription Factor DP1; D014157:Transcription Factors; D001973:Bromodeoxyuridine