Selective alpha v beta 3 integrin blockade potently limits neointimal hyperplasia and lumen stenosis following deep coronary arterial stent injury: evidence for the functional importance of integrin alpha v beta 3 and osteopontin expression during neointima formation.
Lumen loss from vascular restenosis remains a leading cause of chronic revascularization failure.


OBJECTIVE
We hypothesized that cell-matrix adhesion, migration, and differentiation events that underlie restenosis are mediated by alpha v beta 3 integrin-ligand interactions.


METHODS
Using immunohistochemistry and in situ hybridization, we examined the spatial and temporal vessel wall expression of alpha v beta 3 and osteopontin following deep coronary arterial injury. Cell migration and adhesion assays were performed to demonstrate the affinity and specificity of XJ 735 for various vessel wall integrins. The effects of XJ 735 (a selective cyclic Arg-Gly-Asp (RGD) peptidomimetic alpha v beta 3 antagonist) on neointimal hyperplasia and lumen stenosis were tested in a porcine coronary injury model. Normolipemic swine underwent oversized stent injury followed by XJ 735 administration (9 animals, 28 lesions; 1 mg/kg bolus + 7 days 4 mg/kg/d infusion + 21 days 2 mg/kg i.v. bolus 12 hourly) or placebo (10 animals, 30 arterial lesions).


RESULTS
Maximal alpha v beta 3 immunoreactivity was observed between 7-14 days following injury in the neointima, media, and adventitia. Maximal osteopontin mRNA signal in the neointima, media, and adventitia was observed at 14, 7 and 28 days respectively. IC50 for XJ 735 alpha v beta 3-mediated inhibition of human and porcine endothelial cell adhesion, and vascular smooth muscle cell migration, ranged from 0.6 to 4.4 microM. In contrast, IC50 for porcine or human alpha IIb/beta 3, alpha 4 beta 1, alpha v beta 5, and alpha 5 beta 1 inhibition exceeded 100 microM. Steady state XJ 735 plasma levels exceeded 5 microM. Despite slightly higher injury scores in XJ 735 treated animals, significant reductions in mean neointima area (43% reduction; p = 0.0009), and mean percent lumen stenosis (approximately 2.9 fold reduction; p = 0.04) were observed in XJ 735 treated animals. XJ 735 treatment did not significantly alter the relative size of the arterial injury and reference sites (geometric remodeling). Comparison of neontima area vs. injury score regression lines revealed significant reductions in slope (p = 0.0001) and intercept (p = 0.0001) for XJ 735.


CONCLUSIONS
Selective alpha v beta 3 blockade is an effective anti-restenosis strategy that potently limits neointimal growth and lumen stenosis following deep arterial injury. The co-ordinate spatial and temporal upregulation of alpha v beta 3 expression following vessel wall injury, and the high affinity and specificity of XJ 735 for alpha v beta 3, confirms the importance of this integrin in adhesive and migratory cell-matrix events underlying coronary restenosis.
Srivatsa
S S
SS
Department of Internal Medicine, Mayo Clinic, Rochester, MN 55905, USA.
Fitzpatrick
L A
LA
Tsao
P W
PW
Reilly
T M
TM
Holmes
D R
DR
Schwartz
R S
RS
Mousa
S A
SA
D000818:Animals; D002448:Cell Adhesion; D002465:Cell Movement; D003327:Coronary Disease / Q000378:metabolism / Q000517:prevention & control*; D003331:Coronary Vessels / Q000293:injuries* / Q000378:metabolism; D004730:Endothelium, Vascular / Q000378:metabolism; D005340:Fibrinogen / Q000378:metabolism; D053495:Osteopontin; D010456:Peptides, Cyclic / Q000627:therapeutic use*; D011485:Protein Binding; D019030:Receptors, Vitronectin / Q000037:antagonists & inhibitors* / Q000378:metabolism; D012008:Recurrence; D012795:Sialoglycoproteins / Q000378:metabolism; D015607:Stents; D013552:Swine; D019096:Vitronectin / Q000378:metabolism
D010456:Peptides, Cyclic; D019030:Receptors, Vitronectin; C506895:SPP1 protein, human; D012795:Sialoglycoproteins; D019096:Vitronectin; C101857:XJ 735; D053495:Osteopontin; D005340:Fibrinogen