Print Email Facebook Twitter A novel urokinase receptor-targeted inhibitor for plasmin and matrix metalloproteinases suppresses vein graft disease Title A novel urokinase receptor-targeted inhibitor for plasmin and matrix metalloproteinases suppresses vein graft disease Author Eefting, D. Seghers, L. Grimbergen, J.M. de Vries, M.R. de Boer, H.C. Lardenoye, J.W.H.P. Jukema, J.W. van Bockel, J.H. Quax, P.H.A. TNO Kwaliteit van Leven Publication year 2010 Abstract Aims Matrix metalloproteinases (MMP) and plasminogen activator (PA)/plasmin-mediated proteolysis, especially at the cell surface, play important roles in matrix degeneration and smooth muscle cell migration, which largely contributes to vein graft failure. In this study, a novel hybrid protein was designed to inhibit both protease systems simultaneously. MMP and plasmin activity were inhibited at the cell surface by this hybrid protein, consisting of the receptor-binding amino-terminal fragment (ATF) of urokinase-type PA, linked to both the tissue inhibitor of metalloproteinases (TIMP-1) and bovine pancreas trypsin inhibitor (BPTI), a potent protease inhibitor. The effect of overexpression of this protein on vein graft disease was studied. Methods and resultsA non-viral expression vector encoding the hybrid protein TIMP-1.ATF.BPTI was constructed and validated. Next, cultured segments of human veins were transfected with this vector. Expressing TIMP-1.ATF.BPTI in vein segments resulted in a mean 36 ± 14 reduction in neointima formation after 4 weeks. In vivo inhibition of vein graft disease by TIMP-1.ATF.BPTI is demonstrated in venous interpositions placed into carotid arteries of hypercholesterolaemic APOE*3Leiden mice. After 4 weeks, vein graft thickening was significantly inhibited in mice treated with the domains TIMP-1, ATF, or BPTI (36-49 reduction). In the TIMP-1.ATF.BPTI-treated mice, vein graft thickening was reduced by 67±4, which was also significantly stronger when compared with the individual components.Conclusion These data provide evidence that cell surface-bound inhibition of the PA and MMP system by the hybrid protein TIMP-1.ATF.BPTI, overexpressed in distant tissues after electroporation-mediated non-viral gene transfer, is a powerful approach to prevent vein graft disease. © 2010 The Author. Subject HealthBiomedical ResearchGene therapyMatrix metalloproteinasesPlasmin(ogen)Vascular surgeryVein graft diseaseapolipoprotein E3aprotininhybrid proteinmatrix metalloproteinasematrix metalloproteinase inhibitorplasminplasmin inhibitorplasminogen activatortissue inhibitor of metalloproteinase 1tissue inhibitor of metalloproteinase 1 amino terminal fragment of urokinase type plasminogen activator bovine pancreas trypsin inhibitorunclassified drugurokinase receptoranimal cellanimal experimentanimal modelanimal tissuearticlecarotid arterycell surfacecontrolled studyelectroporationexpression vectorfemalegene overexpressiongene targetinggenetic transfectionhumanhuman tissuehypercholesterolemiain vivo cultureintimamalemousenonhumannonviral gene delivery systempriority journalvein graft disease To reference this document use: http://resolver.tudelft.nl/uuid:cb524f30-ee7c-4293-9f93-a07ab74656f4 DOI https://doi.org/10.1093/cvr/cvq203 TNO identifier 425144 ISSN 0008-6363 Source Cardiovascular Research, 88 (2), 367-375 Document type article Files To receive the publication files, please send an e-mail request to TNO Library.