Platelet-derived endothelial cell growth factor gene therapy for limb ischemia.
OBJECTIVE
Platelet-derived endothelial cell growth factor (PD-ECGF) is identical to thymidine phosphorylase (TP), and it can induce angiogenesis, including arteriogenesis, in chronically ischemic canine myocardium. Because its effect on peripheral arterial disease has not been elucidated, we investigated whether overexpression of PD-ECGF/TP could ameliorate chronic limb ischemia in rabbits.


METHODS
Left femoral arteries were resected from 24 male rabbits. After 10 days, a plasmid vector containing human PD-ECGF/TP complimentary DNA was injected into 10 sites in the adductor muscles. Control groups received either the LacZ plasmid vector or saline vehicle only (n = 8 per group). Blood pressure was measured in the calf before surgery, at the onset of ischemia, 10 days later, and 20 and 30 days after gene transfer. Collateral vessel development and limb perfusion were assessed by angiography, and resected tissues underwent molecular and histologic examination.


RESULTS
In the PD-ECGF/TP group, human PD-ECGF/TP messenger RNA and protein were still detected at 30 days after treatment. Calf blood pressure decreased significantly after femoral artery resection in all three groups. It subsequently showed a greater increase in the PD-ECGF/TP group than in either control group, and the difference was significant at 20 days after treatment (PD-ECGF/TP, 97.4 +/- 7.4; LacZ, 58.6 +/- 6.9; saline, 41.3 +/- 3.6). Immunohistochemical staining demonstrated an increased ratio of capillaries and arterioles to muscle fibers in the PD-ECGF/TP group (2.14 +/- 0.13 and 1.51 +/- 0.06), but not in the LacZ group (1.39 +/- 0.04 and 0.71 +/- 0.05) or the saline group (1.34 +/- 0.05 and 0.71 +/- 0.04, P < .01). The angiographic score was higher in the PD-ECGF/TP group (0.96 +/- 0.08) than in the LacZ group (0.50 +/- 0.02) or saline group (0.51 +/- 0.03) at 30 days after gene transfer (P < .01).


CONCLUSIONS
This study demonstrated that PD-ECGF/TP gene transfer induced angiogenesis and decreased ischemia in a rabbit hindlimb model by promoting arteriogenesis, suggesting that targeting this gene may be a promising therapeutic strategy for peripheral vascular disease.
Yamada
Narihisa
N
Second Department of Surgery, Faculty of Medical Sciences, University of Fukui, Fukui, Japan.
Li
Wei
W
Ihaya
Akio
A
Kimura
Tetsuya
T
Morioka
Kouichi
K
Uesaka
Takahiko
T
Takamori
Atsushi
A
Handa
Mitsuteru
M
Tanabe
Sawaka
S
Tanaka
Kuniyoshi
K
D000818:Animals; D001794:Blood Pressure; D004195:Disease Models, Animal; D005121:Extremities / Q000098:blood supply*; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D007511:Ischemia / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology / Q000628:therapy*; D008297:Male; D018482:Muscle, Skeletal / Q000098:blood supply / Q000473:pathology; D018919:Neovascularization, Physiologic; D010957:Plasmids / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism; D011817:Rabbits; D012039:Regional Blood Flow; D013939:Thymidine Phosphorylase / Q000096:biosynthesis* / Q000235:genetics*; D013997:Time Factors; D014162:Transfection
D012333:RNA, Messenger; D013939:Thymidine Phosphorylase