Print Email Facebook Twitter Adenoviral expression of a urokinase receptor - Targeted protease inhibitor inhibits neointima formation in murine and human blood vessels Title Adenoviral expression of a urokinase receptor - Targeted protease inhibitor inhibits neointima formation in murine and human blood vessels Author Quax, P.H.A. Lamfers, M.L.M. Lardenoye, J.H.P. Grimbergen, J.M. de Vries, M.R. Slomp, J. de Ruiter, M.C. Kockx, M.M. Verheijen, J.H. van Hinsbergh, V.W.M. Gaubius Instituut TNO Publication year 2001 Abstract Background - Smooth muscle cell migration, in addition to proliferation, contributes to a large extent to the neointima formed in humans after balloon angioplasty or bypass surgery. Plasminogen activator/plasmin-mediated proteolysis is an important mediator of this smooth muscle cell migration. Here, we report the construction of a novel hybrid protein designed to inhibit the activity of cell surface-bound plasmin, which cannot be inhibited by its natural inhibitors, such as α2-antiplasmin. This hybrid protein, consisting of the receptor-binding amino-terminal fragment of uPA (ATF), linked to the potent protease inhibitor bovine pancreas trypsin inhibitor (BPTI), can inhibit plasmin activity at the cell surface. Methods and Results - The effect of adenovirus-mediated ATF.BPTI expression on neointima formation was tested in human saphenous vein organ cultures. Infection of human saphenous vein segments with Ad.CMV.ATF.BPTI (5 × 109 pfu/mL) resulted in 87.5±3.8% (mean±SEM, n=10) inhibition of neointima formation after 5 weeks, whereas Ad.CMV.ATF or Ad.CMV.BPTI virus had only minimal or no effect on neointima formation. The efficacy of ATF.BPTI in vivo was demonstrated in a murine model for neointima formation. Neointima formation in the femoral artery of mice, induced by placement of a polyethylene cuff, was strongly inhibited (93.9±2%) after infection with Ad.CMV.mATF.BPTI, a variant of ATF.BPTI able to bind specifically to murine uPA receptor; Ad.CMV.mATF and Ad.CMV.BPTI had no significant effect. Conclusions - These data provide evidence that adenoviral transfer of a hybrid protein that binds selectively to the uPA receptor and inhibits plasmin activity directly on the cell surface is a powerful approach to inhibiting neointima formation and restenosis. Chemicals/CAS: Aprotinin, 9087-70-1; Peptide Fragments; Plasmin, EC 220.127.116.11; Recombinant Fusion Proteins; Urinary Plasminogen Activator, EC 18.104.22.168 Subject AdenoviridaeAnimalsAprotininBlood VesselsCattleCHO CellsCricetinaeFemoral ArteryFemoral VeinGene ExpressionGenetic VectorsHumansMiceMice, Inbred C57BLMuscle, Smooth, VascularOrgan Culture TechniquesPeptide FragmentsPlasminRecombinant Fusion ProteinsSaphenous VeinTransfectionTunica IntimaUrinary Plasminogen Activator To reference this document use: http://resolver.tudelft.nl/uuid:95229d60-8b8b-4c4c-b81e-bd6adc9a76ff TNO identifier 235959 ISSN 0009-7322 Source Circulation, 103 (4), 562-569 Document type article Files To receive the publication files, please send an e-mail request to TNO Library.