(Circulation. 2000;102:1308.)
© 2000 American Heart Association, Inc.
Basic Science Reports |
Ribozyme Oligonucleotides Against Transforming Growth Factor-ß Inhibited Neointimal Formation After Vascular Injury in Rat Model
Potential Application of Ribozyme Strategy to Treat Cardiovascular Disease
From the Department of Geriatric Medicine (K.Y., R.M., N.T., H.N., M.A., J.H., T.O.) and the Division of Gene Therapy Science (R.M., Y.K.), Osaka University Medical School, Suita; BILIS (T.S.), Shiga; and Toyohashi Technology University (A.K.), Toyohashi, Japan.
Correspondence to Ryuichi Morishita, MD, PhD, Associate Professor, Division of Gene Therapy Science, Osaka University Medical School, Suita 565, Japan. E-mail morishit{at}geriat.med.osaka-u.ac.jp
 |
Abstract |
|---|
 Top Abstract IntroductionMethodsResultsDiscussionReferences |
|---|
Background—Because the mechanisms of atherosclerosis or restenosis after angioplasty have been postulated to involve an increase in transforming growth factor (TGF)-ß, a selective decrease in TGF-ß may have therapeutic value. Thus, we used the ribozyme strategy to actively cleave the targeted gene to selectively inhibit TGF-ß expression.
Methods and Results—We constructed ribozyme oligonucleotides (ONs) targeted to the sequence of the TGF-ß gene that shows 100% homology among the human, rat, and mouse species. The specificity of ribozyme against TGF-ß gene was confirmed by selective inhibition of TGF-ß mRNA in cultured vascular smooth muscle cells as well as balloon-injured blood vessels in vivo. Importantly, the marked decrease in TGF-ß resulted in significant inhibition of neointimal formation after vascular injury in a rat carotid artery model (P<0.01), whereas DNA-based control ONs and mismatched ribozyme ONs did not have any inhibitory effect on neointimal formation. Inhibition of neointimal formation was accompanied by (1) a reduction in collagen synthesis and mRNA expression of collagen I and III and (2) a significant decrease in DNA synthesis as assessed by proliferating cell nuclear antigen staining. Moreover, we modified ribozyme ONs containing phosphorothioate DNA and RNA targeted to the TGF-ß gene. Of importance, modified ribozyme ONs showed a further reduction in TGF-ß expression.
Conclusions—Overall, this study provides the first evidence that selective blockade of TGF-ß resulted in inhibition of neointimal formation, accompanied by a reduction in collagen synthesis and DNA synthesis in a rat model. We anticipate that modification of ribozyme ON pharmacokinetics will facilitate the potential clinical utility of the ribozyme strategy.
Key Words: atherosclerosis • growth substances • gene therapy • enzymes
|
Introduction |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
As a consequence of the revolutionary developments in the field of molecular biology and their impact on our understanding of the mechanisms of disease processes, treatment strategies that exploit our expanding knowledge of the structures and functions of molecules are being pursued. One strategy for combating disease processes has been to target the transcriptional process. Two approaches have been used to accomplish this. One is the use of antisense oligonucleotides (ONs) complementary to the mRNA of interest.1 The second approach is the use of ribozyme, a unique class of RNA molecules that not only store information but also process catalytic activity.2 Ribozyme is known to catalytically cleave specific target RNA, leading to degradation, whereas antisense ONs inhibit translation by binding to mRNA sequences on a stoichiometric basis. Theoretically, ribozyme is more effective in inhibiting target gene expression. Therefore, we used a novel therapeutic strategy, ribozyme technology, to selectively inhibit target gene expression. Using ribozyme technology, we identified transforming growth factor-ß (TGF-ß) as a target, because TGF-ß plays a pivotal role in the pathogenesis of cardiovascular disease, eg, restenosis after angioplasty.3 4 5 Moreover, TGF-ß increases synthesis of extracellular matrix proteins, including collagen, laminin, and fibronectin, by a variety of cells.6 7 8 From this viewpoint, TGF-ß has been thought to be an ideal target molecule to prevent the progression of cardiovascular disease. Here, using ribozyme TGF-ß ONs, we demonstrated that the selective inhibition of TGF-ß resulted in a significant decrease in neointimal formation after vascular injury in a rat balloon-injury model. This is the first report of successful in vivo gene therapy using ribozyme technology to treat cardiovascular disease. Further innovation of ribozyme technology may open a new era in the treatment of cardiovascular disease.
|
Methods |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
Synthesis of Ribozyme ONs and Selection of Target Sequences
We constructed hammerhead ribozyme ONs targeted against TGF-ß mRNA, as shown in Figure 1
. The target
sequence of TGF-ß was identical among the human, rat, and mouse
species.9 10 11 In this study, we used DNA-based control ONs
as the negative control, because DNA ribozyme has no catalytic
activity. Moreover, mismatched ribozyme ONs that had lost their
catalytic activity were also used as an additional negative
control.
|
In Vivo Gene Transfer
A 2F Fogarty catheter was used to induce vascular injury in male
Sprague-Dawley rats (400 to 500 g; Charles River Breeding
Laboratories, Shizuoka, Japan).12 13 In vivo gene
transfer was performed under the following conditions: vascular injury
of the common carotid artery was induced by the passage and inflation
of a balloon catheter through an arteriotomy in the external carotid
artery 3 times. The injured segment was transiently isolated with
temporary ligatures. Then, 200 µL hemagglutinating virus of Japan
(HVJ)–liposome complex containing either ribozyme ONs, DNA-control
ONs, or mismatched ONs (each at 1 µmol/L contained in liposome)
was incubated within the lumen for 10 minutes at room temperature. At 2
weeks after transfection, each carotid artery was processed for
morphological study.12 13 For histological
analyses, a segment of each artery was perfusion-fixed with 4%
paraformaldehyde at physiological
pressure (110 mm Hg) and subsequently processed. Areas of media
and lumen were measured on a digitizing tablet (Power Laboratory) after
staining with hematoxylin. At least 3 individual sections from the
middle of the transfected arterial segments were
analyzed. Animals were coded so that in the analysis,
the researcher did not know which treatment each individual animal had
received.
Preparation of HVJ-Liposome
We used HVJ-coated liposome, which is reported to provide highly
efficient transfection of cells in culture.12 13 14 15 16 Briefly,
phosphatidylserine, phosphatidylcholine, and
cholesterol were mixed in a weight ratio of 1:4.8:2 to
create a lipid mixture. Purified HVJ (Z strain) was
inactivated by UV irradiation before use. The liposome
suspension was mixed with HVJ, and then free HVJ was removed by sucrose
density gradient centrifugation. This preparation
method has been optimized to achieve maximal transfection efficiency as
reported previously.15
In Vivo Transfection of FITC-Labeled Ribozyme ONs
FITC-labeled ribozyme ONs on the 3' and 5' ends were provided by
Nihon Seifun Inc. Transfer of FITC-labeled ribozyme ONs was performed
according to the following protocol; HVJ complex with FITC-labeled
ribozyme ONs (1 µmol/L) was incubated for 10 minutes. The
vessels were harvested 2 weeks after transfection and perfusion-fixed
with 4% paraformaldehyde. Sections were examined by
fluorescence microscopy after staining in Erichrome black T
solution. Elastic fibers stained dark red and were readily
distinguishable from the specific FITC-labeled ribozyme ONs by
treatment with Erichrome black T solution.17
Northern Blot Analysis
RNA was extracted from injured vessels transfected with ribozyme
ONs (1 µmol/L wrapped in liposome) treated with RNAzol (Tel-Test
Inc) at 1 and 7 days after transfection. Contralateral arteries in
ribozyme ON–transfected animals were also used as intact arteries.
Levels of TGF-ß (1 day after transfection) and collagen I and III (7
days after transfection) mRNA were measured by Northern blot
analysis. The filter was baked, prehybridized, and hybridized
with mouse TGF-ß probe (pMTGFbeta, American Type Culture Collection),
collagen I and III probes (donated by Dr S. Kim, Osaka City
University), and G3PDH probe (Clontech Laboratories, Inc).
Immunohistochemistry
Sections were dewaxed, rehydrated, and incubated with PBS
containing 0.3% hydrogen peroxide to reduce endogenous
peroxidase activity. The sections were then incubated with primary
antibodies or lectin diluted in PBS with 10% horse serum at room
temperature for 60 minutes. After 3 washings in Tris-buffered saline
containing 2% horse serum, species-appropriate biotinylated secondary
antibodies were applied, followed by avidin-biotin peroxidase complex
(Vectastain ABC kit, PK 6100, Vector Laboratories). Omission of primary
antibodies and staining with type- and class-matched irrelevant
immunoglobulin served as a negative control for each antibody. A
monoclonal antibody for collagen I and III (mouse IgG, Chemicon)
recognizes collagen I and III.
Sirius Red Method for Collagen Staining
Sirius red microscopy detects interstitial collagen,
including types I and III.18 19 The stained
sections were observed under polarized light and photographed with the
same exposure time for each section. Analysis of Sirius red
staining was performed with a computer-based quantitative color image
analysis system. Photographs were scanned into a
1000x1000 image buffer of the Optimas 5.2 image
analysis system (Optimas Co). A color threshold mask for
immunostaining was defined to detect the red color by
sampling, and the same threshold was applied to all specimens. The
percentage of the total area with positive color was recorded for
each section.
Quantification of Cell Proliferation in Medial
Lesions
Paraffin-embedded sections of carotid artery harvested 4 days
after transfection were used for quantification of cell
proliferation.7 8 Four sections of each vessel spaced at
0.4-mm intervals were measured by a computerized image analyzer
system (Image Command 5098, Olympus). Monoclonal antibody against
proliferating cell nuclear antigen (PCNA; PC-10; 1:500; DAKO) was used
as a specific marker for proliferating cells. The number of
PCNA-positive nuclei was counted in the medium by the image
analysis system at x400 magnification. Frequency of cell
proliferation was expressed as PCNA index, defined as the ratio of the
number of PCNA-positive nuclei to the total number of nuclei in the
media. Samples were coded so that the analysis was performed
without knowledge of which treatment each individual vessel had
received.
Statistical Analysis
All values are expressed as mean±SEM. ANOVA with subsequent
Bonferroni’s test was used to determine the significance of
differences in multiple comparisons. A value of P<0.05 was
considered statistically significant.
|
Results |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
In Vivo Experiments
To inhibit TGF-ß expression, we constructed "hammerhead" ribozyme ONs targeted to the common sequence of TGF-ß among human, rat, and mouse species (Figure 1
; References 9 through
119 10 11 ). Hammerhead ribozyme is known to cleave the specific
sequences GUH (H=C, U, or A), but not other sequences.2
Therefore, we transfected ribozyme ONs against TGF-ß into a rat
balloon-injury carotid artery model. To enhance the efficiency and
stability, we used the HVJ-liposome delivery system, which is reported
to bypass endocytosis, thereby avoiding degradation in
lysosomes.16 23 Successful transfection of
ribozyme ONs is shown in Figure 2, top
right. Transfection of FITC-labeled ribozyme ONs resulted in
fluorescence in the balloon-injured vessels at 1 day after
transfection. The fluorescence was localized primarily in cell
nuclei (Figure 2) and persisted for up to 2 weeks after
transfection (data not shown). Untreated or HVJ complex without
ON–treated vessels revealed no specific fluorescence (except
autofluorescence) in the elastic lamina, demonstrating that
this fluorescence was specific for FITC-labeled ribozyme
ONs.
|
Given the successful transfection of ribozyme ONs into balloon-injured
vessels, we next examined the in vivo inhibitory effect of
ribozyme TGF-ß ONs on TGF-ß expression induced by vascular injury.
As shown in Figure 1, ribozyme ONs can cleave the mRNA of
TGF-ß at +817, based on the reported sequence,9 10 11
whereas other genes theoretically cannot be cleaved by ribozyme ONs. In
the present study, we used DNA-based control ONs and mismatched
ribozyme ONs as negative controls, because both control ONs have no
catalytic activity. Neither DNA-ribozyme nor mismatched ribozyme ONs
inhibited TGF-ß expression induced by vascular injury (Figure 3). No change in TGF-ß expression was
observed in untransfected blood vessels. In contrast, transfection of
ribozyme TGF-ß ONs significantly decreased TGF-ß expression
(P<0.01, Figure 3). Thus, we examined the effect of
ribozyme TGF-ß ONs on neointimal formation after vascular
injury in the rat carotid artery. As shown in Figure 4, untreated, DNA-based control
ON–transfected, and mismatched ribozyme ON–transfected (1
µmol/L) vessels exhibited neointimal formation at 2 weeks
after transfection. In contrast, a single administration of ribozyme
TGF-ß ONs (1 µmol/L) resulted in a significant reduction in
neointimal formation (P<0.01; Figure
4B). Ribozyme ON treatment did not alter the medial area
(Table). The reduction in
neointimal formation was limited to transfected regions
(data not shown). In addition, treatment with antisense thrombomodulin
oligodeoxynucleotide had no effect on the ratio of
neointimal to medial area as an additional negative control
(data not shown).
|
|
|
It is noteworthy how inhibition of TGF-ß resulted in the blockade of
neointimal formation. Given the previous reports that
TGF-ß stimulated extracellular matrix through the induction of
collagen expression,6 7 8 we postulated that inhibition of
TGF-ß would interfere with the matrix structure. Indeed, we have
observed that mRNAs of collagen I and III were also reduced by ribozyme
TGF-ß ONs, whereas they were induced after vascular injury (Figures 5 and 6).
In contrast, treatment with DNA-based control ONs or mismatched
ribozyme ONs had no effect on collagen mRNA expression (Figures
5 and 6). The reduction of collagen I and III in blood vessels
transfected with ribozyme TGF-ß ONs was also confirmed by the
observation that positive immunohistochemical staining of collagen I
and III was markedly decreased by ribozyme TGF-ß ONs but not
mismatched ribozyme ONs (Figure 7A and 7B). In addition, we performed Sirius red staining for collagen,
because Sirius red staining under polarized light visualizes collagen,
including types I and III.19 Further evidence for the
inhibition of extracellular matrix deposition was provided by the
observation that treatment with ribozyme TGF-ß ONs decreased collagen
content, as assessed by quantitative color image analysis using
Sirius red staining (P<0.01, Figure 8). Blood vessels transfected with
ribozyme TGF-ß ONs showed less positive Sirius red staining,
indicating a low content of interstitial collagen (Figure
8). In contrast, blood vessels transfected with mismatched
ribozyme ONs exhibited substantial accumulation of
interstitial collagen within the neointima
(Figure 8). In addition, as shown in Figure 9A, PCNA-stained nuclei were observed in
the neointimal and medial layers of blood vessels at 4 days
after vascular injury. Transfection of ribozyme ONs resulted in a
significant decrease in the ratio of PCNA-positive nuclei to total
cells at 4 days after transfection compared with blood vessels
transfected with mismatched ribozyme ONs (P<0.01, Figure
9B). In contrast, untransfected intact carotid arteries
exhibited few PCNA-stained nuclei.
|
|
|
|
|
|
Discussion |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
TGF-ß has been of interest in vascular biology because numerous studies indicated it to be increased in cardiovascular disease, such as atherosclerosis and restenosis after angioplasty.3 4 5 Recent studies using gene transfer technology showing that blood vessels expressing the TGF-ß gene develop neointimal formation have focused on the mitogenic action of TGF-ß.24 25 More recently, the blockade of TGF-ß actions through the type II receptor with the dominant-negative construct led to the inhibition of neointimal formation in a rat balloon-injury model.26 However, there are numerous controversial reports that TGF-ß exhibited inhibitory effects on vascular smooth muscle cell (VSMC) proliferation in cultured VSMCs.27 28 29 Thus, it is still unclear whether TGF-ß contributes to the progression of restenotic lesion formation. Therefore, we examined the effects of inhibition of TGF-ß synthesis on neointimal formation after vascular injury. To discuss this issue, we used ribozyme technology rather than antisense, for the following reasons: (1) It is better to target the common sequence of target genes among humans, rats, pigs, etc, for applying results of preclinical animal studies to human trials using antisense or ribozyme technology. However, it is difficult to select antisense sequences around ATG sites that are most effective as antisense sequences, because the structure of the TGF-ß gene around ATG sites is not identified in the human, rat, and mouse species. (2) The inhibitory effect of ribozyme on target gene expression is theoretically greater than that of antisense (passive versus active).2 Fortunately, we detected a possible site as a target sequence for ribozyme catalysis in the TGF-ß gene. As shown in Figure 1
, the target sequences are completely identical among
the human, rat, and mouse species. The present study demonstrated that ribozyme ONs against TGF-ß selectively inhibited TGF-ß production without affecting other genes. The specificity of the inhibitory effect of ribozyme ONs on TGF-ß production presented in this study is supported by several lines of evidence: (1) Ribozyme ONs inhibited TGF-ß expression in vivo, whereas DNA-control ONs and mismatched ONs did not. (2) Ribozyme TGF-ß ONs inhibited TGF-ß but not G3PDH expression, as assessed by Northern blotting in human cultured VSMCs. High transfection efficiency of the HVJ-liposome method into blood vessels has been reported in balloon-injury and vein graft models.12 13 15 16 23 30 31 32 With nucleus-targeted LacZ, transfection of the LacZ gene exhibited diffuse and frequent X-Gal–positive signals in both medial and adventitial layers in vein grafts,30 consistent with a previous report.31 Similar results were also obtained with immunohistochemical staining against nitric oxide in vessels transfected with endothelial constitutive nitric oxide synthase.30 32 In addition, Yonemitsu et al15 documented that HVJ liposomes could achieve highly efficient gene transfection into the medial smooth muscle cells of intact arteries at 150 and 760 mm Hg of pressure (mean=85.3% and 93.5% of total smooth muscle cells, respectively). Moreover, the sufficient transfection efficiency of the HVJ-liposome method to inhibit neointimal formation was also supported by the previous publications.32 33 In contrast, the previous reports documented the expression of TGF-ß in balloon-injured arteries.26 34 35 By in situ hybridization, Smith et al26 reported that proliferating and quiescent smooth muscle cells in denuded vessels expressed high levels of mRNA for TGF-ß. A similar expression pattern of TGF-ß has also been reported.34 35 The localization of transfected cells seems to be identical to that of cells producing TGF-ß.
Using ribozyme ONs against TGF-ß, the present study demonstrated that inhibition of TGF-ß by ribozyme ONs resulted in a significant reduction in neointimal formation in a rat balloon-injury model. How do ribozyme TGF-ß ONs inhibit neointimal formation? As expected, the inhibition of neointimal formation was accompanied by a marked reduction in collagen synthesis and mRNA expression and protein content of collagen I and III, because TGF-ß stimulated extracellular matrix formation. Moreover, ribozyme TGF-ß ONs also significantly reduced DNA synthesis in VSMCs, as assessed by PCNA staining. Although previous reports documented an antiproliferative action of TGF-ß in cultured cells,27 28 29 the in vivo effects of TGF-ß might be mitogenic, consistent with previous reports.24 25 26 These results together suggest that TGF-ß contributes to the restenosis process through the accumulation of collagen synthesis and increase in DNA synthesis of VSMCs rather than antiproliferative actions in vivo. Moreover, this is the first report of successful in vivo application of ribozyme ON technology progressing toward human gene therapy in cardiovascular disease. The practical use of these ribozyme ONs as therapy for atherosclerosis induced by high TGF-ß is dependent on the development of a delivery system into blood vessels. Further studies are necessary to test the efficacy of ribozyme ONs in vivo to examine the therapeutic application. Here, we revealed the first evidence that TGF-ß expression can be selectively prevented by ribozyme ONs directed against the TGF-ß gene, suggesting a novel therapeutic strategy for the treatment of cardiovascular disease related to high TGF-ß. The selective blockade of TGF-ß is particularly attractive, because ribozyme TGF-ß ONs prevented neointimal formation, accompanied by a reduction in collagen synthesis and DNA synthesis in the rat model.
|
Acknowledgments |
|---|
This work was supported in part by grants from the Hoan-sya Foundation and the Japan Cardiovascular Research Foundation, a Japan Heart Foundation Research Grant, a Grant-in-Aid from the Tokyo Biochemical Research Foundation, and a Grant-in-Aid from the Ministry of Education, Science, Sports, and Culture.
Received January 7, 2000; revision received April 7, 2000; accepted April 13, 2000.
|
References |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
1. Flanagan WM. Antisense comes of age. Cancer Metastasis Rev. 1998;17:169–176.[Medline] [Order article via Infotrieve]
2. Kiehntopf M, Esquivel EL, Brach MA, et al. Clinical applications of ribozymes. Lancet. 1995;345:1027–1031.[Medline] [Order article via Infotrieve]
3. Sarzani R, Brecher P, Chobanian AV. Growth factor expression in aorta of normotensive and hypertensive rats. J Clin Invest. 1989;83:1404–1498.
4. Majesky MW, Lindner V, Twardzik DR, et al. Production of transforming growth factor beta-1 during repair of arterial injury. J Clin Invest. 1991;88:904–910.
5. Nikol S, Isner M, Pickering JG, et al. Expression of transforming growth factor-ß1 is increased in human vascular restenosis lesions. J Clin Invest. 1992;90:1582–1592.
6.
Amento EP, Ehsani N, Palmer H, et al.
Cytokines and growth factors positively and negatively regulate
interstitial collagen gene expression in human vascular
smooth muscle cells. Arterioscler Thromb. 1991;11:1223–1230.
7. Zhao Y. Transforming growth factor-beta (TGF-beta) type I and type II receptors are both required for TGF-beta-mediated extracellular matrix production in lung fibroblasts. Mol Cell Endocrinol. 1999;150:91–97.[Medline] [Order article via Infotrieve]
8. Smith WC. TGF beta inhibitors: new and unexpected requirements in vertebrate development. Trends Genet. 1999;15:3–5.[Medline] [Order article via Infotrieve]
9. Derynck R, Jarrett JA, Chen EY, et al. Human transforming growth factor-beta complementary DNA sequence and expression in normal and transformed cells. Nature. 1985;316:701–705.[Medline] [Order article via Infotrieve]
10.
Qian SW, Kondaiah P, Roberts AB, et al. CDNA cloning by
PCR of rat transforming growth factor beta-1. Nucleic Acid
Res. 1990;18:3059.
11.
Derynck R, Jarrett JA, Chen EY, et al. The murine
transforming growth factor-beta precursor. J Biol Chem. 1986;261:4377–4379.
12.
Morishita R, Gibbons GH, Ellison KE, et al. Single
intraluminal delivery of antisense cdc 2 kinase and PCNA
oligonucleotides results in chronic inhibition of
neointimal hyperplasia. Proc Natl Acad Sci
U S A. 1993;90:8474–8478.
13. Morishita R, Gibbons GH, Ellison KE, et al. Intimal hyperplasia after vascular injury is inhibited by antisense cdk 2 kinase oligonucleotides. J Clin Invest. 1994;93:1458–1464.
14.
Kaneda Y, Iwai K, Uchida T. Increased expression of DNA
co-introduced with nuclear protein in adult rat liver.
Science. 1989;243:375–378.
15. Yonemitsu Y, Kaneda Y, Morishita R, et al. Characterization of in vivo gene transfer into the arterial wall mediated by the Sendai virus (hemagglutinating virus of Japan) liposomes: an effective tool for the in vivo study of arterial diseases. Lab Invest. 1996;75:313–323.[Medline] [Order article via Infotrieve]
16. Morishita R, Gibbons GH, Horiuchi M, et al. Molecular delivery system for antisense oligonucleotides: enhanced effectiveness of antisense oligonucleotides by HVJ-liposome mediated transfer. J Cardiovasc Pharmacol Ther. 1996;2:213–222.
17. Kittelberger R, Davis PF, Stehbens WE. An improved immunofluorescence technique for the histological examination of blood vessel tissue. Acta Histochem. 1989;86:137–142.[Medline] [Order article via Infotrieve]
18. Janqueira LCU, Bignolas G, Brentani RR. Picrosirius staining plus polarization microscopy, a specific method for collagen detection in tissue sections. Histochem J. 1979;11:447–455.[Medline] [Order article via Infotrieve]
19.
Aikawa M, Rabkin E, Okada Y, et al. Lipid lowering by
diet reduces matrix metalloproteinase activity and increases collagen
content of rabbit atheroma: a potential mechanism of lesion
stabilization. Circulation. 1998;97:2433–2444.
22. Morishita R, Gibbons GH, Horiuchi M, et al. Role of AP-1 complex in angiotensin II-mediated transforming growth factor-ß expression and growth of smooth muscle cells: using decoy approach against AP-1 binding site. Biochem Biophys Res Commun. 1998;243:361–367.[Medline] [Order article via Infotrieve]
23. Morishita R, Gibbons GH, Kaneda Y, et al. Pharmacokinetics of antisense oligonucleotides (cyclin B1 and cdc 2 kinase) in the vessel wall: enhanced therapeutic utility for restenosis by HVJ-liposome method. Gene.. 1994;149:13–19.[Medline] [Order article via Infotrieve]
24. Wolf YG, Rasmussen LM, Ruoslahti E. Antibodies against transforming growth factor-ß1 suppress intimal hyperplasia in a rat model. J Clin Invest. 1994;93:1172–1178.
25.
Nabel EG, Shum L, Pompili VJ, et al. Direct transfer of
transforming growth factor beta 1 gene into arteries stimulates
fibrocellular hyperplasia. Proc Natl Acad Sci U S A. 1993;90:10759–10763.
26.
Smith JD, Bryant SR, Couper LL, et al. Soluble
transforming growth factor-beta type II receptor inhibits negative
remodeling, fibroblast transdifferentiation, and intimal lesion
formation but not endothelial growth. Circ
Res. 1999;84:1212–1222.
27. Gibbons GH, Pratt RE, Dzau VJ. Vascular smooth muscle cell hypertrophy vs. hyperplasia: autocrine transforming growth factor-beta 1 expression determines growth response to angiotensin II. J Clin Invest. 1992;90:456–461.
28.
Koibuchi Y, Lee WS, Gibbons GH, et al. Role of
transforming growth factor-beta 1 in the cellular growth response to
angiotensin II. Hypertension. 1993;21:1046–1050.
29.
Stouffer GA, Owens GK. Angiotensin
II–induced mitogenesis of spontaneously hypertensive rat–derived
cultured smooth muscle cells is dependent on autocrine
production of transforming growth factor-ß. Circ
Res. 1992;70:820–828.
30. Matsumoto T, Komori K, Yonemitsu Y, et al. Hemagglutinating virus of Japan-liposome mediated gene transfer of endothelial cell nitric oxide synthase inhibits intimal hyperplasia of canine vein grafts under conditions of poor runoff. J Vasc Surg. 1998;27:125–144.
31.
Morishita R, Gibbons GH, Kaneda Y, et al. Novel in
vitro gene transfer method for study of local modulators in vascular
smooth muscle cells. Hypertension.. 1993;21:894–899.
32.
Leyen HVL, Gibbons GH, Morishita R, et al. Gene therapy
inhibiting neointimal vascular lesion: in vivo transfer of
ec-nitric oxide synthase gene. Proc Natl Acad Sci U S A. 1995;92:1137–1141.
33. Bai HZ, Morishita R, Kida I, et al. Inhibition of intimal hyperplasia after vein grafting by in vivo transfer of human senescent cell-derived inhibitor-1 gene. Gene Ther. 1998;5:761–769.[Medline] [Order article via Infotrieve]
34. Majesky MW, Lindner V, Twardzik DR, et al. Production of transforming growth factor beta 1 during repair of arterial injury. J Clin Invest. 1991;88:904–910.
35. Grant MB, Wargovich TJ, Bush DM, et al. Expression of IGF-1, IGF-1 receptor and TGF-beta following balloon angioplasty in atherosclerotic and normal rabbit iliac arteries: an immunocytochemical study. Regul Pept. 1999;79:47–53.[Medline] [Order article via Infotrieve]
This article has been cited by other articles:
 |
|
 |
|
  E.-H. Yao, N. Fukuda, T. Ueno, H. Matsuda, H. Nagase, Y. Matsumoto, H. Sugiyama, and K. Matsumoto A pyrrole-imidazole polyamide targeting transforming growth factor-{beta}1 inhibits restenosis and preserves endothelialization in the injured artery Cardiovasc Res, March 1, 2009; 81(4): 797 - 804. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Rodriguez, J. Martinez-Gonzalez, B. Raposo, J. F. Alcudia, A. Guadall, and L. Badimon Regulation of lysyl oxidase in vascular cells: lysyl oxidase as a new player in cardiovascular diseases Cardiovasc Res, July 1, 2008; 79(1): 7 - 13. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Khan, A. Agrotis, and A. Bobik Understanding the role of transforming growth factor-{beta}1 in intimal thickening after vascular injury Cardiovasc Res, May 1, 2007; 74(2): 223 - 234. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Bobik Transforming Growth Factor-{beta}s and Vascular Disorders Arterioscler Thromb Vasc Biol, August 1, 2006; 26(8): 1712 - 1720. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Wang, J. Zhang, G. Spinetti, L.-Q. Jiang, R. Monticone, D. Zhao, L. Cheng, M. Krawczyk, M. Talan, G. Pintus, et al. Angiotensin II Activates Matrix Metalloproteinase Type II and Mimics Age-Associated Carotid Arterial Remodeling in Young Rats Am. J. Pathol., November 1, 2005; 167(5): 1429 - 1442. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Kobayashi, K. Yokote, M. Fujimoto, K. Yamashita, A. Sakamoto, M. Kitahara, H. Kawamura, Y. Maezawa, S. Asaumi, T. Tokuhisa, et al. Targeted Disruption of TGF-{beta}-Smad3 Signaling Leads to Enhanced Neointimal Hyperplasia With Diminished Matrix Deposition in Response to Vascular Injury Circ. Res., April 29, 2005; 96(8): 904 - 912. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Ikeda, Y. Imai, H. Kumagai, T. Nosaka, Y. Morikawa, T. Hisaoka, I. Manabe, K. Maemura, T. Nakaoka, T. Imamura, et al. Vasorin, a transforming growth factor {beta}-binding protein expressed in vascular smooth muscle cells, modulates the arterial response to injury in vivo PNAS, July 20, 2004; 101(29): 10732 - 10737. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J.-M. Li, L. M. Fan, A. Shah, and G. Brooks Targeting {alpha}v{beta}3 and {alpha}5{beta}1 for gene delivery to proliferating VSMCs: synergistic effect of TGF-{beta}1 Am J Physiol Heart Circ Physiol, August 7, 2003; 285(3): H1123 - H1131. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. H. Von der Thusen, J. Kuiper, T. J. C. Van Berkel, and E. A. L. Biessen Interleukins in Atherosclerosis: Molecular Pathways and Therapeutic Potential Pharmacol. Rev., March 1, 2003; 55(1): 133 - 166. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Kotani, N. Fukuda, H. Ando, W.-Y. Hu, S. Kunimoto, S. Saito, and K. Kanmatsuse Chimeric DNA-RNA hammerhead ribozyme targeting PDGF A-chain mRNA specifically inhibits neointima formation in rat carotid artery after balloon injury Cardiovasc Res, January 1, 2003; 57(1): 265 - 276. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
U. Kintscher, C. Lyon, S. Wakino, D. Bruemmer, X. Feng, S. Goetze, K. Graf, A. Moustakas, B. Staels, E. Fleck, et al. PPAR{alpha} Inhibits TGF-{beta}-Induced {beta}5 Integrin Transcription in Vascular Smooth Muscle Cells by Interacting With Smad4 Circ. Res., November 29, 2002; 91 (11): e35 - e44. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. C. Lowe, S. N. Oesterle, and L. M. Khachigian Coronary in-stent restenosis: Current status and future strategies J. Am. Coll. Cardiol., January 16, 2002; 39(2): 183 - 193. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. A. Kingston, S. Sinha, A. David, M. G. Castro, P. R. Lowenstein, and A. M. Heagerty Adenovirus-Mediated Gene Transfer of a Secreted Transforming Growth Factor-{beta} Type II Receptor Inhibits Luminal Loss and Constrictive Remodeling After Coronary Angioplasty and Enhances Adventitial Collagen Deposition Circulation, November 20, 2001; 104(21): 2595 - 2601. [Abstract] [Full Text] [PDF]
|
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||