(Circulation. 1999;99:3199-3205.)
© 1999 American Heart Association, Inc.
Basic Science Reports |
Expression of Tissue Inhibitor of Matrix Metalloproteinases 1 by Use of an Adenoviral Vector Inhibits Smooth Muscle Cell Migration and Reduces Neointimal Hyperplasia in the Rat Model of Vascular Balloon Injury
From the Hatter Institute, University College London Hospitals (C.M.D., J.R.M.) and the Division of Cardiovascular Genetics, Department of Medicine, University College London (S.E.H.), London, UK; Genetic Therapy Inc, Gaithersburg, Md (A.M.); and the Department of Molecular Pathology, University College London Medical School (D.S.L.), London, UK.
Correspondence to Dr Clare M. Dollery, Department of Cardiology, 4th Floor Jules Thorn Building, The Middlesex Hospital, London, W1N8AA, UK. E-mail c.dollery{at}ucl.ac.uk
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Abstract |
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Background—Cell migration is a major contributor to injury-induced neointimal hyperplasia and depends on alteration of the proteolytic balance within the arterial wall toward matrix breakdown. This is partly mediated by the matrix metalloproteinases (MMPs) and their natural inhibitors, the tissue inhibitors of metalloproteinases (TIMPs).
Methods and Results—An increase in expression of biologically active and immunoreactive TIMP-1 was seen in vitro after infection of rat smooth muscle cells (SMCs) with Av1.TIMP1 (an adenoviral vector containing the human TIMP1 cDNA). Infection of rat SMCs with Av1.TIMP1 reduced migration in vitro by 27% compared with control virus–infected cells (37.6±4.34 versus 51±5.01 cells per high-power field, P<0.05). The adenoviral vector was delivered to the injured rat carotid artery, and 4 days later, immunoreactive protein was identified and migration of SMCs reduced by 60% (5.2±0.5 versus 12.8±1.5 cells per section, P<0.05, n=5). Neointimal area 14 days after injury showed a 30% reduction in the animals receiving the Av1.TIMP1 virus compared with controls (0.09±0.01 versus 0.14±0.01 mm2, P=0.02, n=14).
Conclusions—The response to arterial balloon injury involves MMP-dependent SMC migration and can be attenuated in vivo by the transmural expression of TIMP-1 by adenoviral gene transfer.
Key Words: restenosis • genes • muscle, smooth • cells
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Introduction |
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Extracellular matrix stability is important in maintaining the integrity of the vessel wall, and recent studies have focused on its role in the postprocedural vasculopathies, such as angioplasty restenosis.1 2 3 4 The restenotic process after vascular injury is a result of normal but unwanted healing causing lumen loss and return of symptoms in 25% of patients.5
After arterial injury, vascular smooth muscle cells (VSMCs) proliferate and migrate, forming a neointima that later accumulates extracellular matrix.6 VSMCs secrete proteases, which facilitate migration by plasminogen-dependent and -independent pathways.1 Expression of both plasminogen activators and inhibitors after rat arterial balloon injury correlates with migration of VSMCs.7 Vascular injury has been shown to cause early upregulation of both gelatinase A and B,1 3 4 but the tissue inhibitors of the matrix metalloproteinases (MMPs), the TIMPs, appear to be expressed 48 to 72 hours after the peak of the MMPs.1 3 4 It is ultimately the balance between the MMPs and their inhibitors that determines the focal proteolysis around the SMCs of the arterial media. Thus, in the first 1 to 4 days after balloon injury, a rapid alteration of the proteolytic balance toward matrix breakdown occurs concurrently with proliferation and migration of SMCs. This process is established by 7 days after injury, when TIMP2 and plasminogen activator inhibitor-1 are upregulated.4 8 The later phase of the arterial response to injury is characterized by the cessation of SMC migration and commencement of matrix accumulation.6
It was our hypothesis in this study that adenoviral gene transfer of the cDNA encoding TIMP immediately after injury would result in increased levels of TIMP1 and its earlier expression. This rapid inhibition of matrix breakdown might then reduce the early phases of SMC migration and attenuate neointima formation. An adenoviral vector was used because it allows efficient direct in vivo gene transfer to quiescent native SMCs. Accordingly, the purpose of this study was to investigate the effects of increased TIMP1 expression on proliferation and migration of SMCs both in vitro and in vivo within the arterial wall.
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Methods |
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Generation of Adenovirus Vectors
Two adenoviral vectors were used, Av1.LacZ4 and Av1.TIMP1. Av1.LacZ4 is a replication-deficient first-generation adenoviral vector encoding ß-galactosidase.9 The cDNA encoding human TIMP1 was the kind gift of Prof Gillian Murphy (University of East Anglia). The TIMP1 cDNA was excised from pBluescript TIMP1 with BamH1 and HindIII, and the restriction sites were filled with Klenow fragment. The blunt-ended fragment was cloned into the EcoRV-digested pAVS6A vector.9 Correct insertion and orientation were confirmed by restriction enzyme analysis and sequence analysis. Recombinant viruses were generated by calcium phosphate cotransfection of pAVS6A TIMP1 with the ClaI fragment of dl 327 adenovirus type 5 on 293 cells. Recombinant adenovirus clones were screened for the TIMP1 cDNA by Southern analysis and plaque-purified. Viral stocks were generated in 293 cells and concentrated by cesium chloride ultracentrifugation. The resultant bands were dialyzed against 10 mmol/L Tris (pH 7.4), 1 mmol/L MgCl2, and 10% glycerol. Plaque titration on 293 cells showed titers of 1x1010 and 1x1011 pfu/mL for Av1.LacZ4 and Av1.TIMP1, respectively.
Cell Culture and Adenovirus Infection
Primary VSMCs were isolated from male Wistar rats and used at
passage 4 to 6.10 Cells were incubated at 37°C in a
humidified 5% CO2/95% O2
incubator in RPMI medium supplemented with 10% FCS, 100 U/mL
penicillin, 100 U/mL streptomycin, and 1% transferrin.
Immunohistochemistry using an anti–
-smooth muscle actin antibody
(Sigma) confirmed staining in 98% of cells. Eighty percent confluent
monolayers were transferred to serum-free RPMI 24 hours before
infection. Cells were incubated with Av1.TIMP1 or Av1.LacZ4 for 1 hour.
The infection media were then replaced with serum-free RPMI.
Conditioned media harvested 48 hours later were concentrated by use of
Centricon-3 (Amicon Inc) filters and assayed for protein
content.11
Western Blot Analysis and Gelatin and Reverse
Zymography
For Western analysis, protein was loaded and run on a
10% SDS-polyacrylamide gel and transferred to nitrocellulose
(Amersham). Mouse anti–human TIMP-1 monoclonal antibody (the kind gift
of Dr A. Docherty, Celltech Ltd) was added at 1:1000 for 2 hours and
probed with 1:1000 horseradish peroxidase–conjugated rabbit anti-mouse
monoclonal antibody (Sigma). Enzyme-linked chemiluminescence
detection was used to visualize the signal (Amersham). MMP and TIMP
activities were assayed by gelatin and reverse zymography,
respectively, as previously described.4 Briefly, samples
in nonreducing buffer were run on a 10% gel containing 1 mg/mL of
gelatin (Sigma) for zymography plus 0.1 mL of hamster gelatinase A
solution/mg gelatin for reverse zymography (UTI Inc). Gels were washed
in 2.5% Triton X-100 and incubated for 16 hours at 37°C before
staining with Coomassie blue.
Invasion Assay and Proliferation of SMCs In Vitro
Biocoat Matrigel invasion chambers were used according to the
manufacturer's instructions for invasion studies (Becton Dickinson).
Briefly, SMCs were infected as described above and 24 hours later were
trypsinized and seeded into cell culture inserts at a density of
2x106/mL in serum-free RPMI (Gibco). The cells
conditioned the surrounding media for 24 hours before the insert was
placed in a companion plate containing 300 ng/mL platelet-derived
growth factor (R&D Systems). Forty-eight hours later, membranes
were fixed and stained by use of the Prodiff Kit (Baxter). Filters were
viewed with a Labphot 2A microscope (Nikon) and images digitized with a
JVC-TK1281 video camera and Lucia-M image analysis system.
Invasion was quantified by counting the mean number of cells per 3
high-power fields per chamber.
DNA synthesis was assayed by incorporation of [3H]thymidine (1 µCi/mL) into quiescent and proliferating SMCs. SMCs infected as described above were allowed to proliferate for 48 hours before pulsing with [3H]thymidine. Some cells were stimulated to proliferate with either 10 ng/mL basic fibroblast growth factor (bFGF) or 20% FCS. SMCs were precipitated with 10% trichloroacetic acid and dissolved in 0.5 mol/L NaOH. A scintillation counter was used to quantify [3H]thymidine incorporation and normalized to the total protein content per well to correct for unequal seeding density.11
Animal Surgery and In Vivo Gene Delivery
Male Wistar rats (350 to 400 g) were anesthetized,
and the left common carotid artery was balloon injured with a 2F
embolectomy catheter inserted via the external carotid artery as
previously described.4 Heparin (35 IU) was then
administered systemically via the external jugular vein before
isolation of a 1-cm length of the common carotid artery with
arterial clips. A 2F cannula was then inserted into the
common carotid artery and irrigated with 0.9% saline. Viral infusion
mixtures containing 5x108 pfu of Av1.LacZ4 or
Av1.TIMP1, diluted to a total volume of 100 µL in RPMI, were then
incubated in the arterial lumen for 20 minutes, removed,
and flow in the artery was restored.
Vessel Harvesting
Vessels were harvested at 2, 4, and 14 days after gene transfer.
Tissues were fixed by 10 minutes of retrograde perfusion at 120
mm Hg via an aortic cannula. Four percent formaldehyde or 2%
formaldehyde/0.2% glutaraldehyde were the fixatives
used for immunohistochemistry or histomorphometry, respectively. In a
subset of animals, fresh tissues were frozen in liquid nitrogen, and
protein was extracted for Western analysis of human TIMP1 as
described above.4
Immunohistochemistry and Assessment of Proliferation
Arteries were embedded in paraffin and cut into 5-µm sections.
Sections were stained with a mouse extravidin-peroxidase detection
system (Sigma) and diaminobenzidine (Sigma Fast DAB) according to the
manufacturer's instructions. Sections harvested at 2 and 4 days were
incubated for 1 hour with 1:200 anti–human TIMP1 antibody.
Proliferation was assessed 2 and 14 days after injury with rabbit
polyclonal anti–Ki 67 antibody (1:25) incubated for 1 hour and light
hematoxylin counterstaining. The cell number within the media and
neointima was quantified 4 days after injury by counting
nuclei in 3 hematoxylin and eosin (H+E)–stained sections at 100-µm
intervals.
Vessel Morphometry, Collagen Studies, and Migration of SMCs In
Vivo
Sections were stained with H-E and examined with the microscope
and image analysis system described above. Areas and ratios
were determined by tracing the intimal, medial, and adventitial
boundaries. Collagen studies used picrosirius red staining to compare
collagen content by calculating the integral density of the staining in
the media and neointima of each section.12
Migration of VSMCs was assessed by counting the cells within the
internal elastic lamina 4 days after gene transfer. Serial sections
were stained with Verhoeff–van Gieson stain to locate the internal
elastic lamina. In all measurements, the observer was blinded to the
treatment group from which the sections came.
Statistical Analysis
All values are expressed as mean±SEM. Student's t
test was used to examine the difference between experimental
groups.
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Results |
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Expression and Activity of TIMP-1 In Vitro
Expression of the transgene human TIMP1 in rat VSMCs infected with Av1.TIMP1 was demonstrated by Western analysis of conditioned media and compared with media from uninfected cells or cells infected with a control virus (Av1.LacZ4) (Figure 1
). Immunoreactive TIMP1 increased with
the multiplicity of infection (MOI) of the virus. Human TIMP1 was seen
to inhibit rat gelatinases (data not shown). The biological activity of
the TIMP1 was demonstrated by reverse zymography (Figure 2). In addition to a major band of
activity at 28 kDa in Av1.TIMP1-infected cells, the gels also showed
smaller bands at the same molecular weight in controls. These bands are
likely to represent constitutively secreted rat TIMP-1, which
was not detected by Western analysis for human TIMP1. At higher
MOIs of Av1.TIMP1, a lower-molecular-weight band was seen by both
Western analysis and reverse zymography; this is likely to be
partially glycosylated human TIMP1.
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TIMP1 Does Not Affect DNA Synthesis in VSMCs but Inhibits Migration
Across a Basement Membrane Barrier
No significant change in DNA synthesis was seen in
Av1.TIMP1-infected cells compared with uninfected or Av1.LacZ4-infected
cells (all, P=NS) (Figure 3).
In contrast, cells stimulated with bFGF and infected with higher MOIs
of Av1.TIMP1 (5, 50, and 100) showed significantly greater
proliferation than cells infected with 100 MOI of Av1.LacZ4 (all,
P<0.05). Comparison of Av1.TIMP1-infected cells stimulated
with FGF with the more appropriate control of uninfected FGF-stimulated
cells showed no significant difference. The Av1.LacZ4-infected cells
showed reduced DNA synthesis compared with control cells in both
quiescent and stimulated conditions, as previously
reported.13 The invasion assay revealed a 27%
reduction in migration through a Matrigel barrier in cells infected
with Av1.TIMP1 at an MOI of 100 compared with Av1.LacZ4 (Figure 4) (P<0.05, n=4).
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Presence of Human TIMP1 Protein After In Vivo Gene
Transfer
Successful transgene expression in vivo was demonstrated by
widespread transmural human TIMP1 immunostaining, which
was greatest in the media (Figure 5). No
immunoreactivity was seen in arteries infected with Av1.LacZ4. Western
analysis of arterial extracts for human TIMP1
confirmed a 28-kDa band in Av1.TIMP1-infected arteries that was absent
in Av1.LacZ4-infected arteries (Figure 5).
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Av1.TIMP1 Infection Inhibits Neointimal Hyperplasia
In Vivo
The effect of TIMP1 expression on neointimal
hyperplasia was quantified by histomorphometric analysis of
arterial cross sections from 28 animals 14 days after
vascular injury. A 32% reduction in neointimal area in the
Av1.TIMP1-infected arteries was seen compared with Av1.LacZ4-infected
arteries (0.14±0.01 and 0.09±0.01 mm2 in
the Av1.LacZ4 and Av1.TIMP1 groups, respectively, P=0.02)
(Figure 6). Similarly, the ratio of
neointima to media was 27% less in Av1.TIMP1 arteries
compared with Av1.LacZ4 (P=0.04) (Figure 6). To
assess changes in overall vessel wall size, the area within the
external elastic lamina (EEL) was calculated and showed a mean area of
0.448±0.02 cm2 in the Av.LacZ4 group and
0.403±0.01 cm2 in the Av1.TIMP1 group
(P=NS).
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Role of DNA Synthesis and Cell Number, SMC Migration, and Matrix
Accumulation in Reducing Neointimal Hyperplasia
The relative contributions of cellular migration and proliferation
and matrix accumulation in the observed reduction in
neointimal hyperplasia were assessed. Ki 67
immunoreactivity 2 days after balloon injury showed no significant
difference in the Av1.TIMP1-infected arteries compared with the
Av1.LacZ4 at either time point (Ki 67 labeling index: day 2, 19.5±5.3
versus 19.2±3.3, P=NS; day 14, 0.35±0.16 versus
0.22±0.10, P=NS). The total cell number within the EEL 4
days after injury in the Av1.TIMP1-infected arteries was 40% less than
in the Av1.LacZ4-treated arteries, suggesting significant cell loss
(260±36 compared with 434±56, P=0.048, n=5). Previous
studies in our laboratory show that medial cell number in uninjured
artery of animals 72 to 96 hours after contralateral
arterial injury does not differ from Av1.LacZ4-treated
arteries nor from sham-operated, saline-irrigated vessels (data not
shown).
In vivo migration was assessed by counting all cells within the
internal elastic lamina in 20 sections taken at 100-µm intervals in
each artery. A highly significant difference of 5.2±0.5 versus
12.8±1.5 cells per section was seen between the Av1.TIMP1 and
Av1.LacZ4 arteries (P=5x10-6)
(Figure 7).
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Collagen content was assessed in 14-day lesions by densitometric analysis of picrosirius red–stained sections. The integral density of picrosirius red staining in the neointima and media of Av1.TIMP1-infected arteries was significantly lower than in Av1.LacZ4 arteries (2483±171 versus 3161±187, P=0.01). The apparent decrease in total collagen staining in the Av1.TIMP1 arteries is likely simply to reflect the smaller neointimal area of the arteries. When the picrosirius red staining was expressed per cell, the results were not significantly different (Av1.TIMP1, 2.14±0.13 per cell versus Av1.LacZ4, 2.12±0.17 per cell).
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Discussion |
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The results presented show that in vivo arterial gene transfer of TIMP1 attenuates neointimal hyperplasia after vascular injury, with a marked reduction in SMC migration but without altering proliferation. It is our hypothesis that the proteolytic state induced by vascular injury was prevented by increased levels of TIMP1 protein demonstrated by Western analysis and immunohistochemistry. In vitro experiments showed that secretion of biologically active TIMP1 after Av1.TIMP1 infection inhibits VSMC migration, and this was confirmed by a 60% reduction in migration of SMCs in vivo. Both in vivo and in vitro analysis of DNA synthesis and proliferation showed no reduction in proliferation in response to TIMP1, and in stimulated cells in vitro, TIMP1 was mildly promitogenic. This study confirms that in vivo gene transfer of TIMP1 alters neointimal formation and presents the first in vivo evidence that attenuation of the response to vascular injury by direct gene transfer of TIMP1 alters SMC migration.
Our in vitro studies largely parallel the findings of Forough et al,14 who used retroviral vectors to express TIMP1 in SMCs that were transferred to the arterial wall. This technique prevents examination of migration of SMCs, because the arterial lumen is colonized by transferred cells. In contrast, the efficiency of adenoviral vectors allows direct in vivo gene transfer and yields high levels of episomal transgene expression, which is well suited to temporary genetic modification of postprocedural vasculopathy.
Our results show a 27% reduction in in vitro invasion of SMCs through a basement membrane barrier when infected with Av1.TIMP1 compared with control or Av1.LacZ4 cells. This is consistent with findings in retrovirally transduced cells.14 Inhibition of invasion by TIMPs has been demonstrated in cancer and angiogenesis.15 16 Migration of SMCs in vivo is difficult to quantify, but 2 methods have been described.1 17 Both techniques rely on the observation that SMCs first appear in the neointima 3 to 4 days after injury and that 96% of these cells result from migration and not proliferation.17 18 We counted intimal cells in 20 transverse sections taken at 100-µm intervals along the length of infected arteries and observed a 60% reduction in Av1.TIMP1-infected arteries compared with those infected with Av1.LacZ4.
We saw no evidence of an alteration in DNA synthesis in quiescent cells after Av1.TIMP1 infection, but cells stimulated with bFGF showed a small increase compared with Av1.LacZ4-infected cells but not with uninfected cells. The control vector, Av1.LacZ4, reflects the effects of adenoviral infection, which are unrelated to TIMP1 gene expression.19 TIMP1 is known to be a mitogen in some cell types.20 In our experiments, however, no increase in proliferation was observed compared with uninfected cells. Interestingly, retroviral gene transfer of TIMP1 in vitro is associated with a reduction in cell number compared with cells infected with a control retrovirus.14 Cell number, however, may be influenced by both the rate of DNA synthesis and loss of cells by necrosis or apoptosis.
The Ki 67 antibody reacts with cells at all stages of the cell cycle but not with cells in the G0 phase.21 With this antibody, no difference was detected between the Av1.LacZ4 and Av1.TIMP1 groups. This is not unexpected, because <10% of cells in the arterial wall are infected by this method of gene transfer. This result reflects predominantly a comparison between cells exposed to TIMP1 protein and uninfected cells: these exhibit similar proliferation in vitro. There was, however, a decrease in cell number in the media and neointima of the Av1.TIMP1-infected arteries 4 days after vascular injury. Neither in vitro nor in vivo results suggest that this is due to altered SMC proliferation. Av1.TIMP1-infected arteries, however, show a lower cell number than control groups. Adenoviral gene transfer can cause loss of medial cells at high titer but would have been detected in the Av1.LacZ4 group and would have increased neointima formation.19
There is a significant cell loss in the arteries from the Av1.TIMP1-treated group. This may be due to necrosis or apoptosis. It is known that extracellular matrix contact can mediate cell survival and that an imbalance in the matrix composition of the arterial wall might mediate the cell loss observed.22 TIMP3 has recently been shown to cause apoptosis in VSMCs, but TIMP1 does not appear to share this property in vitro.23
The inhibition of the MMP enzymes is likely to result in early-onset matrix accumulation after vascular injury. However, no constrictive remodeling could be identified when the areas subtended by the EEL in Av1.TIMP1 and Av1.LacZ4 arteries were compared. We used a modification of the method of Coats et al12 to investigate the collagen content of vessels 14 days after gene transfer. Integral density of the media and neointima of picrosirius red–stained sections was measured, reflecting the collagen content. Unexpectedly, this showed more collagen in the Av1.LacZ4-treated group than in the Av1.TIMP1 group. However, when these results were normalized to the number of cells in the artery, the results showed no difference, suggesting the same cell-to-matrix ratio in both groups. These results reflect those found when a nonspecific pharmacological MMP inhibitor (GM6001) was used in the rabbit injury model, in which collagen content was reduced by MMP inhibition.6 In the rat, GM6001 reduced in vivo SMC migration by 97% 4 days after injury, but the final lesion was unchanged at 14 days.2 This has been explained by a phase of "catch-up" growth between 10 and 14 days after injury. One interpretation of these studies was that inhibiting SMC migration was not sufficient to inhibit lesion growth.
Our results show that in vivo transfer of the TIMP1 gene inhibits
migration of SMCs and reduces neointimal hyperplasia 14
days after injury. This supports previous analysis of the rat
model suggesting that 50% of the neointimal cells are
derived through proliferation and 50% through migration and may
explain the incomplete inhibition of neointimal hyperplasia
by inhibition of either proliferation or migration
alone.24 The difference between pharmacological
inhibitors and these studies of gene transfer may be due to
the specificity or level of inhibition of the MMPs. In contrast to the
studies discussed above, 1 report using a broad-spectrum MMP
inhibitor, batimastat, in the rat carotid model showed
reduced neointimal formation. This may be because
batimastat is a less specific MMP inhibitor and may inhibit
other metalloproteinases, such as the disintegrin and metalloproteinase
(ADAMs) group, whose role as a TNF-–converting enzyme has been
investigated.25 We chose to transfer TIMP1 because,
although it inhibits all the MMPs, it has a particular specificity for
gelatinase B, which is upregulated soon after vascular injury. The
important difference in this gene transfer approach to vascular injury
may be the reduction in cell number in the arterial wall
after TIMP1 gene transfer. TIMP1 does not appear to cause frank
apoptosis or necrosis in vitro but might interact with other
matrix components in vivo, such as tenascin C, which are known to
mediate cell survival. If the change in cell number is due to a
specific biological effect of TIMP1 on either necrotic or programmed
cell death, it may influence the application of this vector not only in
vascular injury but also in tumor metastasis.
In summary, this study shows that direct in vivo gene transfer of TIMP1 using an adenoviral vector inhibits neointimal development by 30% and that a similar reduction in SMC migration both in vivo and in vitro may be the underlying mechanism. We have also shown a reduction in cell number in the arterial wall after TIMP1 gene transfer in vivo. In this study, the use of TIMP gene transfer provides evidence of the role of MMPs in vascular SMC migration and the response to vascular injury. To date, the majority of gene transfer studies of the response to arterial balloon injury have focused on reducing proliferation of SMCs, but our results suggest that migration of these cells has an equally important role. Ultimately, the unwanted aspects of the response to vascular injury may be best targeted by inhibition of both migration and proliferation of SMCs. The MMPs and their inhibitors are a major regulatory component in these responses.
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Acknowledgments |
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Dr Dollery was supported by a Clinical Training Fellowship awarded by the Medical Research Council of Great Britain and by the Jean Shanks Foundation. Work in the laboratory of Dr McEwan is funded by The British Heart Foundation. We are grateful to Michelle Rollence and Dr Sue Stevenson of Genetic Therapy Inc for their work in the recombination of the Av1.TIMP1 vector and to Dr Karen Webb for preparing the TIMP1 adenoviral shuttle vector.
Received December 11, 1998; revision received February 1, 1999; accepted March 16, 1999.
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References |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
1. Bendeck MP, Zempo N, Clowes AW, Galardy RE, Reidy MA. Smooth muscle cell migration and matrix metalloproteinase expression after arterial injury in the rat. Circ Res. 1994;75:539–545.
2.
Bendeck MP, Irvin C, Reidy MA. Inhibition of matrix
metalloproteinase activity inhibits smooth muscle cell migration but
not neointimal thickening after arterial
injury. Circ Res. 1996;78:38–43.
3.
Southgate KM, Fisher M, Banning AP, Thurston VJ, Baker
AH, Fabunmi RP, Groves PH, Davies M, Newby AC. Upregulation of basement
membrane-degrading metalloproteinase secretion after balloon injury of
pig carotid arteries. Circ Res. 1996;79:1177–1187.
4.
Webb KE, Henney AM, Anglin S, Humphries SE, McEwan J.
The expression of matrix metalloproteinases and their
inhibitor TIMP-1 in the rat carotid artery carotid after
balloon injury. Arterioscler Thromb Vasc Biol. 1997;17:1837–1844.
5.
Landau C, Lange RA, Hillis LD.
Percutaneous transluminal coronary angioplasty.
N Engl J Med. 1994;330:981–993.
6.
Strauss BH, Robinson R, Batchelor WB, Chisholm RJ,
Ravi G, Natarajan MK, Logan RA, Mehta SR, Levy DE, Ezrin AM, Keeley FW.
In vivo collagen turnover following experimental balloon angioplasty
injury and the role of matrix metalloproteinases. Circ Res. 1996;79:541–550.
7.
Reidy MA, Irvin C, Lindner V. Migration of
arterial wall cells: expression of plasminogen
activators and inhibitors in injured rat
arteries. Circ Res. 1996;78:405–414.
8.
Hasenstab D, Forough R, Clowes AW.
Plasminogen activator inhibitor
type 1 and tissue inhibitor of metalloproteinases-2
increase after arterial injury in rats. Circ
Res. 1997;80:490–496.
9. Smith TAG, Mehaffey MG, Kayda DB, Saunders JM, Yei S, Trapnell BC, McClelland A, Kaleko M. Adenovirus-mediated expression of therapeutic plasma levels of human factor IX in mice. Nat Genet. 1993;5:397–402.[Medline] [Order article via Infotrieve]
10. Rennick R, Campbell JH, Campbell GR. Vascular smooth muscle cell phenotype and growth behaviour can be influenced by macrophages in vitro. Atherosclerosis. 1988;71:35–43.[Medline] [Order article via Infotrieve]
11. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein using the principle of protein dye binding. Anal Biochem. 1976;72:248–254.[Medline] [Order article via Infotrieve]
12.
Coats WD Jr, Whittaker P, Cheung DT, Currier JW, Han B,
Faxon DP. Collagen content is significantly lower in restenotic
versus nonrestenotic vessels after balloon angioplasty in the
atherosclerotic rabbit model. Circulation. 1997;95:1293–1300.
13.
Yang ZY, Simari RD, Perkins ND, San H, Gordon D, Nabel
GJ, Nabel EG. Role of the p21 cyclin-dependent kinase
inhibitor in limiting intimal cell proliferation in
response to arterial injury. Proc Natl Acad Sci
U S A. 1996;93:7905–7910.
14.
Forough R, Koyama N, Hasenstab D, Lea H, Clowes M,
Nikkari ST, Clowes AW. Overexpression of tissue inhibitor
of matrix metalloproteinase-1 inhibits vascular smooth muscle cell
functions in vitro and in vivo. Circ Res. 1996;79:812–820.
15.
Anand Apte B, Pepper MS, Voest E, Montesano R, Olsen B,
Murphy G, Apte SS, Zetter B. Inhibition of angiogenesis by tissue
inhibitor of metalloproteinase-3. Invest Ophthalmol
Vis Sci. 1997;38:817–823.
16. Matsuzawa K, Fukuyama K, Hubbard SL, Dirks PB, Rutka JT. Transfection of an invasive human astrocytoma cell line with a TIMP-1 cDNA: modulation of astrocytoma invasive potential. J Neuropathol Exp Neurol. 1996;55:88–96.[Medline] [Order article via Infotrieve]
17.
Jackson CL, Raines EW, Ross R, Reidy MA. Role of
endogenous platelet-derived growth factor in
arterial smooth muscle cell migration after balloon
catheter injury. Arterioscler Thromb. 1993;13:1218–1226.
18. Clowes AW, Clowes MM, Fingerle J, Reidy MA. Kinetics of cellular proliferation after arterial injury, V: role of acute distension in the induction of smooth muscle proliferation. Lab Invest. 1989;60:360–364.[Medline] [Order article via Infotrieve]
19. Newman KD, Dunn PF, Owens JW, Schulick AH, Virmani R, Sukhova G, Libby P, Dichek DA. Adenovirus-mediated gene transfer into normal rabbit arteries results in prolonged vascular cell activation, inflammation, and neointimal hyperplasia. J Clin Invest. 1995;96:2955–2965.
20. Hayakawa T, Yamashita K, Ohuchi E, Shinagawa A. Cell growth-promoting activity of tissue inhibitor of metalloproteinases-2 (TIMP-2). J Cell Sci. 1994;107:2373–2379.[Abstract]
21. Leers MP, Theunissen PH, Ramaekers FC, Schutte B. Multi-parameter flow cytometric analysis with detection of the Ki67-Ag in paraffin embedded mammary carcinomas. Cytometry. 1997;27:283–289.[Medline] [Order article via Infotrieve]
22. Khwaja A, Rodriguez Viciana P, Wennstrom S, Warne PH, Downward J. Matrix adhesion and Ras transformation both activate a phosphoinositide 3-OH kinase and protein kinase B/Akt cellular survival pathway. EMBO J. 1997;16:2783–2793.[Medline] [Order article via Infotrieve]
23. Baker AH, Zaltsman AB, George SJ, Newby AC. Divergent effects of tissue inhibitor of metalloproteinase-1, -2, or -3 overexpression on rat vascular smooth muscle cell invasion, proliferation, and death in vitro: TIMP-3 promotes apoptosis. J Clin Invest. 1998;101:1478–1487.[Medline] [Order article via Infotrieve]
24.
Clowes AW, Schwartz SM. Significance of quiescent
smooth muscle migration in the injured rat carotid artery. Circ
Res. 1985;56:139–145.
25. Black RA, Rauch CT, Kozlosky CJ, Peschon JJ, Slack JL, Wolfson MF, Castner BJ, Stocking KL, Reddy P, Srinivasan S, Nelson N, Boiani N, Schooley KA, Gerhart M, Davis R, Fitzner JN, Johnson RS, Paxton RJ, March CJ, Cerretti DP. A metalloproteinase disintegrin that releases tumour-necrosis factor-alpha from cells. Nature. 1997;385:729–733.[Medline] [Order article via Infotrieve]
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