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
Health
Biomedical Research
Gene therapy
Matrix metalloproteinases
Plasmin(ogen)
Vascular surgery
Vein graft disease
apolipoprotein E3
aprotinin
hybrid protein
matrix metalloproteinase
matrix metalloproteinase inhibitor
plasmin
plasmin inhibitor
plasminogen activator
tissue inhibitor of metalloproteinase 1
tissue inhibitor of metalloproteinase 1 amino terminal fragment of urokinase type plasminogen activator bovine pancreas trypsin inhibitor
unclassified drug
urokinase receptor
animal cell
animal experiment
animal model
animal tissue
article
carotid artery
cell surface
controlled study
electroporation
expression vector
female
gene overexpression
gene targeting
genetic transfection
human
human tissue
hypercholesterolemia
in vivo culture
intima
male
mouse
nonhuman
nonviral gene delivery system
priority journal
vein 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