Title
Gene transfer of the urokinase-type plasminogen activator receptor-targeted matrix metalloproteinase inhibitor TIMP-1.ATF suppresses neointima formation more efficiently than tissue inhibitor of metalloproteinase-1
Author
Lamfers, M.L.M.
Grimbergen, J.M.
Aalders, M.C.
Havenga, M.J.
de Vries, M.R.
Huisman, L.G.M.
van Hinsbergh, V.W.M.
Quax, P.H.A.
Gaubius instituut TNO
Publication year
2002
Abstract
Proteases of the plasminogen activator (PA) and matrix metalloproteinase (MMP) system play an important role in smooth muscle cell (SMC) migration and neointima formation after vascular injury. Inhibition of either PAs or MMPs has previously been shown to result in decreased neointima formation in vivo. To inhibit both protease systems simultaneously, a novel hybrid protein, TIMP-1.ATF, was constructed consisting of the tissue inhibitor of metalloproteinase-1 (TIMP-1) domain, as MMP inhibitor, linked to the receptor-binding amino terminal fragment (ATF) of urokinase. By binding to the u-PA receptor this protein will not only anchor the TIMP-1 moiety directly to the cell surface, it will also prevent the local activation of plasminogen by blocking the binding of urokinase-type plasminogen activator (u-PA) to its receptor. Adenoviral expression of TIMP-1.ATF was used to inhibit SMC migration and neointima formation in human saphenous vein segments in vitro. SMC migration was inhibited by 65% in Ad.TIMP-1.ATF-infected cells. Infection with adenoviral vectors encoding the individual domains, Ad.TIMP-1 and Ad.ATF, reduced migration by 32% and 52%, respectively. Neointima formation in saphenous vein organ cultures infected with Ad.TIMP-1.ATF was inhibited by 72% compared with 42% reduction after Ad.TIMP-1 infection and 34% after Ad.ATF infection. These data show that binding of TIMP-1.ATF hybrid protein to the u-PA receptor at the cell surface strongly enhances the inhibitory effect of TIMP-1 on neointima formation in human saphenous vein cultures.
Subject
Biology
Adenovirus
Matrix metalloproteinases
Neointima formation
Plasmin(ogen)
Urokinase-type plasminogen activator
Adenovirus
Adenoviridae
Animals
Cell Division
Cell Membrane
Cell Movement
Cells, Cultured
CHO Cells
Collagenases
Cricetinae
Culture Media, Conditioned
Enzyme Activation
Flow Cytometry
Gene Transfer Techniques
Humans
Matrix Metalloproteinase 13
Muscle, Smooth, Vascular
Protein Structure, Tertiary
Receptors, Cell Surface
Recombinant Fusion Proteins
RNA, Messenger
Saphenous Vein
Tissue Inhibitor of Metalloproteinase-1
Tunica Intima
Urinary Plasminogen Activator
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DOI
https://doi.org/10.1161/01.res.0000041418.51906.57
TNO identifier
236777
ISSN
0009-7330
Source
Circulation Research, 91 (10), 945-952
Document type
article