PMID- 20562095
OWN - NLM
STAT- MEDLINE
DCOM- 20110126
LR  - 20121115
IS  - 1755-3245 (Electronic)
IS  - 0008-6363 (Linking)
VI  - 88
IP  - 2
DP  - 2010 Nov 1
TI  - A novel urokinase receptor-targeted inhibitor for plasmin and matrix 
      metalloproteinases suppresses vein graft disease.
PG  - 367-75
LID - 10.1093/cvr/cvq203 [doi]
AB  - 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 
      RESULTS: A 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.
FAU - Eefting, Daniel
AU  - Eefting D
AD  - Department of Vascular Surgery, Leiden University Medical Center, PO Box 9600, 
      2300 RC Leiden, The Netherlands.
FAU - Seghers, Leonard
AU  - Seghers L
FAU - Grimbergen, Jos M
AU  - Grimbergen JM
FAU - de Vries, Margreet R
AU  - de Vries MR
FAU - de Boer, Hetty C
AU  - de Boer HC
FAU - Lardenoye, Jan-Willem H P
AU  - Lardenoye JW
FAU - Jukema, J Wouter
AU  - Jukema JW
FAU - van Bockel, J Hajo
AU  - van Bockel JH
FAU - Quax, Paul H A
AU  - Quax PH
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20100617
PL  - England
TA  - Cardiovasc Res
JT  - Cardiovascular research
JID - 0077427
RN  - 0 (Apolipoprotein E3)
RN  - 0 (Matrix Metalloproteinase Inhibitors)
RN  - 0 (Receptors, Urokinase Plasminogen Activator)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (Tissue Inhibitor of Metalloproteinase-1)
RN  - 9087-70-1 (Aprotinin)
RN  - EC 3.4.21.7 (Fibrinolysin)
RN  - EC 3.4.21.73 (Urokinase-Type Plasminogen Activator)
RN  - EC 3.4.24.- (Matrix Metalloproteinases)
SB  - IM
MH  - Animals
MH  - Apolipoprotein E3/genetics
MH  - Aprotinin/biosynthesis/genetics
MH  - Atherosclerosis/genetics/metabolism/pathology
MH  - Carotid Arteries/surgery
MH  - Cattle
MH  - Cell Line
MH  - *Cell Proliferation
MH  - Disease Models, Animal
MH  - Electroporation
MH  - Fibrinolysin/antagonists & inhibitors/*metabolism
MH  - *Genetic Therapy/methods
MH  - Graft Occlusion, Vascular/genetics/metabolism/pathology/*prevention & control
MH  - Humans
MH  - Hypercholesterolemia/genetics/metabolism/pathology
MH  - Hyperplasia
MH  - Male
MH  - Matrix Metalloproteinase Inhibitors
MH  - Matrix Metalloproteinases/*metabolism
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - Mice, Mutant Strains
MH  - Receptors, Urokinase Plasminogen Activator/*metabolism
MH  - Recombinant Fusion Proteins/biosynthesis
MH  - Saphenous Vein/*metabolism/pathology/surgery
MH  - Time Factors
MH  - Tissue Culture Techniques
MH  - Tissue Inhibitor of Metalloproteinase-1/biosynthesis/genetics
MH  - Transfection
MH  - Urokinase-Type Plasminogen Activator/biosynthesis/genetics
MH  - Venae Cavae/*metabolism/pathology/transplantation
EDAT- 2010/06/22 06:00
MHDA- 2011/01/28 06:00
CRDT- 2010/06/22 06:00
PHST- 2010/06/22 06:00 [entrez]
PHST- 2010/06/22 06:00 [pubmed]
PHST- 2011/01/28 06:00 [medline]
AID - cvq203 [pii]
AID - 10.1093/cvr/cvq203 [doi]
PST - ppublish
SO  - Cardiovasc Res. 2010 Nov 1;88(2):367-75. doi: 10.1093/cvr/cvq203. Epub 2010 Jun 
      17.