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(Circulation. 2003;107:1046.)
© 2003 American Heart Association, Inc.

Basic Science Reports

Temporal Response and Localization of Integrins ß1 and ß3 in the Heart After Myocardial Infarction

Regulation by Cytokines

Mei Sun, MD, PhD; M. Anne Opavsky, MD, PhD; Duncan J. Stewart, MD; Marlene Rabinovitch, MD; Fayez Dawood, DVM; Wen-Hu Wen, MD; Peter P. Liu, MD

From the Heart and Stroke/Richard Lewar Centre of Excellence and Division of Cardiology, University Health Network (M.S., F.D., W.-H.W., P.P.L); Research Institute, The Hospital for Sick Children (M.A.O., M.R.); and Division of Cardiology, St Michael’s Hospital (D.J.S.), University of Toronto, Canada.

Correspondence to Dr Peter Liu, Heart and Stroke/Richard Lewar Centre of Excellence, EN12-324, 200 Elizabeth St, Toronto, Ontario, M5G 2C4, Canada. E-mail peter.liu{at}utoronto.ca

	Abstract

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Background— Integrins are involved in structural remodeling and tissue repair. This study aimed to elucidate the role of the ß-integrins in cardiac remodeling after myocardial infarction (MI).

Methods and Results— The MI model was created by ligation of the left anterior descending coronary artery in rats. We detected cardiac integrins ß1 and ß3 gene expression (quantitative in situ hybridization) and protein production (Western blot and immunohistochemistry) and potential regulation by tumor necrosis factor (TNF) using neonatal ventricular myocytes and TNF^-/- knockout mice. Integrins ß1 and ß3 gene expression and protein production were low in sham-operated hearts. After MI, the ß1 and ß3 mRNA and proteins were significantly increased at the site of MI at day 3, reached a peak at day 7, and gradually declined thereafter. Integrin ß1A localized primarily in fibroblasts and inflammatory cells, ß1D localized in myocytes, and integrin ß3 was associated primarily with endothelial and smooth muscle cells in peri-infarct vessels. In cultured myocytes, there was isoform transition from the adult ß1D to the fetal ß1A on exposure to TNF-. This was confirmed in vivo in the peri-infarct myocytes, but the transition was voided in TNF^-/--knockout mice.

Conclusions— Integrins ß1 and ß3 are significantly activated in the infarcted myocardium. Integrin ß1 is active particularly at sites of inflammation and fibrosis, whereas integrin ß3 localizes to vessels in the peri-infarct zone in a temporally coordinated manner. Integrin ß1D to ß1A isoform transition in myocytes is regulated by TNF-.

Key Words: myocardial infarction • integrins • remodeling • angiogenesis • tumor necrosis factor-

	Introduction

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Integrins are a family of transmembrane heterodimeric receptors that are widely expressed on the cell surface and provide a physical and biochemical bridge between components of the extracellular matrix and the intracellular physiological environment. They are composed of and ß subunits that individually consist of a large extracellular domain, a transmembrane region, and a relatively short cytoplasmic domain for signaling.^1–4

Integrins mediate cell–cell and cell–matrix interactions in response to stress that leads to intracellular signal transduction, cytoskeletal rearrangements, wound healing, and cell proliferation, differentiation, and death, processes important in tissue repair.⁵ Integrin expression is also associated with changes in extracellular matrix, coordinating synthesis of collagen, fibronectin, and angiogenesis.^6,7

After myocardial injury such as infarction (MI), extensive remodeling takes place both in myocytes and in the extracellular matrix. Adverse remodeling can set the stage for ventricular dysfunction and heart failure. During remodeling, the extracellular domains of existing integrins can be shed into the extracellular space, and new isoforms can be reexpressed on the cell surface to rapidly generate de novo cell–matrix connections.⁸ Previous work has demonstrated significant changes in several -integrin subunits (1, 2, and 5) after MI.⁹ ß1 integrins may play an important coordinating role in extracellular matrix synthesis and remodeling, as demonstrated in skin and lung after injury.⁵ Another potentially important role of integrins in the heart is their ability to serve as mechanotransducers during development and in response to physiological and pathological signals.^10–13 ß1 integrin comes in 2 isoforms in the heart: ß1A, the fetal isoform, responsible for cardiac morphogenesis and plasticity, and ß1D, the adult isoform, important for facilitation of contractility.¹⁴ Integrin ß3 is essential for smooth muscle and endothelial cell migration^15,16 and blood vessel formation in granulation tissue.¹⁷ Currently, the potential role of ß1 and ß3 in post-MI remodeling remains uncertain, and whether ß1 isoforms can be regulated by inflammation and cytokines in the myocytes is unknown.

In the present study, we sought to determine (1) the temporal expression and spatial localization of integrins ß1 and ß3 in the infarcted heart and (2) whether the functional isoforms of ß1 integrins can be regulated by cytokines such as tumor necrosis factor (TNF)-.

	Methods

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Animals
Left ventricular MI was created in 12-week-old male Sprague-Dawley rats (Charles River Canada, Quebec, Canada) by ligation of the left anterior descending coronary artery as previously described by our laboratory.¹⁸ Animals were randomized into sham-operated controls (n=7) and infarcted animals that were randomly euthanized on postoperative days 3, 7, 14, and 28 (n=7 per time point). Hearts were immediately harvested, frozen, and stored at -80°C.

To examine the role of TNF on integrin ß1 isoform expression in vivo, TNF-knockout mice (TNF^-/-) and wild-type controls (WT) (C57BL/6-TNFtm1Gk1, Jackson Laboratory, Bar Harbor, Me) were also subjected to ligation of the left anterior descending coronary artery similar to the protocol outlined above.

For details of methodology on cell culture and in situ hybridization, please refer to the online-only Data Supplement.

Generation of Integrin ß1D, ß1A Polyclonal Antibody
Rabbit polyclonal anti-peptide antibodies against the 17-mer CPINNFKNPNYGRKAGL of ß1D integrin, which contains a unique amino acid sequence present only in the ß1D but not the ß1A isoform, were generated. In addition, rabbit polyclonal anti-peptide antibodies against the 12-amino-acid sequence (CTTVVNPKYEGK) of ß1A integrin, which are isoform specific and do not cross-react with ß1D and other known ß subunit variants, as previously documented, were generated.¹⁹ Rabbits were immunized subcutaneously with 1 mg of the respective conjugate and boosted 5 weeks later with 0.5 mg of the same conjugate. Pooled antiserum was affinity-purified and stored at 4°C in the presence of 2% BSA. Antibody specificity was confirmed by Western blot.

Western Blot Analysis
Protein was extracted from the freshly prepared tissues/cells with lysis buffer after homogenization. It was diluted 1:1 with 2x SDS sample buffer (Invitrogen Novex). Equal amounts of protein (40 µg) were loaded in each lane of 8% to 16% Tris-glycin gel (Helixx). Proteins were separated by electrophoresis and transferred from the gel to a nitrocellulose membrane with an electroblotting apparatus. Membranes were first incubated with 5% BSA for 1 hour and then incubated with anti-integrin ß1, ß3 (PharMingen), ß1A, and ß1D overnight at 4°C. After incubation, samples were washed and subsequently incubated with peroxidase-conjugated secondary antibody and detected by use of the ECL detection Kit (Amersham).

Immunohistochemistry
Cryostat sections (5 µm) were prepared, air-dried, and fixed in cold acetone (-20°C) for 10 minutes. The endogenous peroxidase activity was blocked by 0.3% hydrogen peroxide and incubated with 10% normal mouse/goat serum. Reaction with primary antibody was performed overnight at 4°C. After a washing in PBS, secondary antibodies, IgG-peroxidase–conjugated goat anti-mouse polyclonal antibody (PharMingen) for ß3 and goat anti-rabbit biotinylated antibody (Vector) for ß1D and ßA were used. Bound antibodies were detected with streptavidin-peroxidase complex with DAB. Negative control sections were incubated with secondary antibody alone.

Reverse Transcription–Polymerase Chain Reaction
Total RNA was extracted from cultured myocytes. RT-PCR was performed with primers for ß1, ß3, 1, v, 7, and GAPDH (Table). With PCR buffers and Taq1 polymerase, the cycle parameters were as follows: denaturation at 95°C for 2 minutes, annealing at 60°C for 1 minute, and extension at 72°C for 2 minutes for 33 cycles with 5 minutes of final elongation at 75°C. Reaction mixture without template cDNA was used as a negative control.

View this table: [in this window] [in a new window]   Oligonucleotide Primers

Statistical Analysis
Statistical analyses of in situ hybridization and Western blot findings were performed by ANOVA (SAS). Values are expressed as mean±SEM, with P<0.05 considered significant.

	Results

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Cardiac Pathology
After coronary ligation, myocardial necrosis in the ligated zone was evident on day 3, with sarcolemmal disruption and nucleus dropout. By day 7, inflammatory cells were observed in large numbers in the peri-infarct region. New blood vessel formation and fibroblast proliferation were also evident at the site of infarction. On day 14, inflammatory infiltration subsided, and necrotic tissue was partially replaced by fibroblast-like cells. By day 28, necrotic tissue was completely replaced with fibrotic tissue.

Myocardial Gene Expression of Integrin ß1
With in situ hybridization (Figure 1), we observed low levels of expression of integrin ß1 mRNA in both ventricles of the sham-operated hearts. After MI, integrin ß1 was markedly increased at the site of MI at day 3, with particularly robust expression in the peri-infarct border zone. Integrin ß1 mRNA levels in the infarcted myocardium reached a peak at day 7. At days 14 and 28, integrin ß1 mRNA gradually declined at the site of infarction but remained significantly higher than in the contralateral wall. (Quantitative analysis of ß1 and ß3 is available online.)

View larger version (67K): [in this window] [in a new window]   Figure 1. In situ hybridization detection of integrins ß1 in sham and infarcted rat hearts. Integrin ß1 mRNA was markedly increased in peri-infarct area of MI on day 3, reached a peak on day 7, and gradually declined on days 14 and 28.

Localization of Integrin ß1 in the Normal and Infarcted Myocardium
To determine whether change in integrin ß1 mRNA transcripts results in actual changes in integrin protein, immunostaining with specific integrin ß1A and ß1D antibodies was performed (Figure 2). In sham-operated rat heart, interstitial fibroblasts, vascular smooth muscle cells, and endothelial cells were labeled with anti-integrin isoform ß1A antibody (A), whereas myocytes were labeled only by antibodies specific to integrin isoform ß1D (B). After MI, within the infarct zone, integrin ß1A was localized primarily to the inflammatory cells, blood vessels, and fibroblast-like cells at days 3 and 7. At days 14 and 28, integrin ß1A was localized primarily in fibroblasts (C, E, G, I). Integrin ß1D was not found in nonmyocyte tissue at any time (D, F, H, J).

View larger version (94K): [in this window] [in a new window]   Figure 2. Cells expressing integrin isoforms ß1A and ß1D in normal and infarcted rat myocardium. In normal myocardium, integrin ß1A was labeled in interstitial fibroblasts (red arrow), vascular smooth muscle cells, and endothelial cell (black arrow) (A), whereas ß1D was expressed only in myocytes (B). After MI, integrin ß1A was seen clearly in inflammatory cells (IF) at site of MI on days 3 and 7 (C, E) but in fibroblasts (FB) on day 28 (I). Integrin ß1D was expressed only in myocytes in infarcted heart (D, F, J). G and H show higher magnification. (Magnification: G and H, x400; other panels, x200.)

Changes in protein production of the total integrin ß1 were also quantified by Western blot. The integrin ß1 isoforms do not display the standard mature and immature forms as distinct bands. At all time points, tissue of the infarcted model showed an increase in total integrin ß1 levels similar to the pattern that was observed for gene expression (Figure 3). The highest level of integrin ß1 protein was found in the infarct as well as on day 3: 7 peri-infarct zones. Contralateral zones ß1 levels exhibited no significant change.

View larger version (41K): [in this window] [in a new window]   Figure 3. Western blot analysis for total integrin ß1 (ß1A+ß1D) and ß3 in sham-operated and infarcted rat heart. Integrin ß1 and ß3 from infarcted, peri-infarct, and contralateral zones on days 3, 7, 14, and 28 after MI.

Myocardial Integrin ß3 Gene Expression
Expression of the integrin ß3 mRNA is shown in Figure 4. By in situ hybridization, low levels of integrin ß3 mRNA were observed uniformly in both ventricles of sham-operated heart. After MI, integrin ß3 followed a pattern similar to integrin ß1, in which the mRNA was largely increased at the site of MI at day 3, with particular concentration at the border zone between the infarcted and noninfarcted regions. Integrin ß3 mRNA levels in the infarcted myocardium reached a peak at day 7. At days 14 and 28, integrin ß3 mRNA gradually declined. Compared with sham, integrin ß3 mRNA levels were unchanged in noninfarcted myocardium.

View larger version (91K): [in this window] [in a new window]   Figure 4. In situ hybridization detection of integrin ß3 in rat heart. Integrin ß3 mRNA density was increased in infarct zone on day 3 after MI, reached a peak by day 7, and gradually declined thereafter (A).

Localization of Integrin ß3 in the Normal and Infarcted Myocardium
As shown in Figure 5, the immunohistochemistry localized ß3 primarily to blood vessels and interstitial cells (A). At early stages after MI, new vessels within the peri-infarct and infarct zones were strongly stained for ß3, whereas inflammatory cells and fibroblast-like cells at the site of MI were, by contrast, only minimally stained (B, C). At days 14 and 28, interstitial cells expressed very low levels of integrin ß3, whereas blood vessels still expressed high levels of integrin ß3 (D).

View larger version (153K): [in this window] [in a new window]   Figure 5. Immunohistochemical detection of integrin ß3 expression in rat heart. In sham-operated heart, myocytes and vessels were found expressing integrin ß3 (A). In infarcted heart, blood vessels (arrow) expressed high integrin ß3 levels on day 7 (B, C), whereas fibroblast-like cells were found expressing integrin ß3 in later stages (D). Magnification: A and C, x400; B and D, x200.

Regional production of integrin ß3 was evaluated quantitatively by Western blot (Figure 3). Integrin ß3 protein showed low levels in the normal myocardium after MI. Its content in the infarct and peri-infarct myocardium was significantly increased at day 3 and remained elevated over the course of the 4-week observation. Compared with controls, integrin ß3 levels in noninfarcted myocardium remained unchanged at all time points. (Quantitative data available online.)

Effect of TNF on Integrin ß1 Gene Expression
Our previous studies showed that TNF is increased within hours after MI, with upregulation particularly in the infarct and peri-infarct zones,¹⁸ following a pattern very similar to that observed for integrin ß1. To determine whether TNF may regulate integrin ß1 expression in myocytes, we isolated and cultured neonatal myocytes and exposed them to TNF- over different periods of time. Integrin mRNA expression over the time course is illustrated in Figure 5. Most interestingly, when myocytes in culture were exposed to TNF 12 hours later, there was a time-dependent decrease in integrin ß1D expression but no change in integrin ß1A. To determine whether this is unique to ß1, we also assessed ß3, 1, 3, v, and 7 mRNA expression. But in all cases, TNF did not have an effect on any other integrin isoforms. The transcription levels of integrins ß1A and ß1D protein production were also demonstrated in Western blot (Figure 6).

View larger version (72K): [in this window] [in a new window]   Figure 6. Effect of TNF- on integrin ß1 isoform (ß1A and ß1D) mRNA and protein expression in cultured rat neonatal myocytes. After cells were exposed to TNF for 1, 3, 6, 12, 16, or 24 hours, total RNA was extracted to perform RT-PCR for ß1, ß3, 1, v, 7, and GAPDH (A). RT-PCR for integrin ß1 produced lower and upper bands representing ß1A and ß1D, respectively. Proteins were isolated for Western blot detection of integrin ß1A and ß1D (B). Bands represented here are from 1 of 4 experiments.

To confirm that the mechanisms observed above are also relevant in vivo, we performed MI in mice homozygous for TNF^-/- or their WT littermates as controls. We have found that in the WT animals, in which there is a significant increase in local TNF production in the peri-infarct zone, integrin ß1D was detected at relatively low levels in the myocytes of the peri-infarct zone (Figure 7A, d and e). In the TNF^-/- animals, however, the level of ß1D was high in the corresponding region (Figure 7A, f). This suggested that there is also downregulation of ß1D by TNF in the peri-infarct zone. No integrin ß1A was expressed in myocytes (Figure 7A, a–c). Western blot analysis showed that the integrin ß1D expression increased in TNF^-/- at the site of the peri-infarct zone after MI day 7 but was decreased in WT mice. In contrast, the integrin ß1A increased in both WT and TNF^-/- mice (Figure 7, B and C).

View larger version (100K): [in this window] [in a new window]   Figure 7. Immunostaining for integrin ß1A and integrin ß1D in heart of WT and TNF knockout mice (KO). Integrin ß1A was negatively labeled in myocytes of WT and KO mice (a–c). Integrin ß1D was expressed in myocytes in both WT and KO mice (d–f), but KO mice showed stronger labeling than WT at day 7 in peri-infarct zone (e, f). Magnification: a and d, x200; b, c, e, f, x1000. B, Western blot analysis for integrin ß1D and ß1A in peri-infarcted zone of mouse heart at day 7 after MI: sham (lane 1), KO (lane 2), and WT (lane 3). C, Band values (mean±SEM). *P<0.05.

	Discussion

Top Abstract Introduction Methods Results Discussion References

 
Cell–cell and cell–extracellular matrix interactions provide a structural, chemical, and mechanical substrate that is essential for regulation of cellular phenotype in specific environments. Integrins provide the critical interface between the cell and the matrix and may serve as a candidate mechanotransducer during cardiac development or in response to injury during pathophysiological stress.^12,13,20 In the present study, we observed that both integrins ß1 and ß3 gene expression were temporally upregulated after MI. In particular, ß1D is associated with myocytes, ß1A with interstitial and inflammatory cells, and ß3 with new vascular structures. ß1D is downregulated rapidly by cytokines such as TNF.

We demonstrated that upregulation of integrin ß1A expression is synchronous with other changes in the extracellular matrix, including collagen, fibronectin, osteopontin, tenacin, and others.²¹ The changes in integrins are probably important to allow new connections to be made between the remaining viable cells and the new matrix in the peri-infarct zone, similar to those seen in models of hypertrophy.^22–24

Recently, changes of -integrins after MI have been documented. By day 7 after MI, the 1 integrin expression was elevated both in the remaining normal myocytes in the peri-infarct zone and in the remodeled tissue in the infarct zone.⁹

In our study, the early rise of integrin ß1 production in the repairing myocardium suggests its potential participatory role in the local inflammatory process. Our immunohistochemical study reveals that inflammatory cells contribute to integrin ß1A production in the infarcted myocardium. Cells expressing integrin ß1A at the site of MI are primarily fibroblasts and inflammatory cells, whereas those positive in integrin ß1D are primarily myocytes. These findings suggest that integrin ß1 plays a role in inflammation and subsequent fibrosis, similar to the processes of tissue repair observed in liver,²⁵ lung,^26,27 and kidney.²⁸

Studies have demonstrated that a variety of growth factors upregulate the expression of several integrins. These growth factors, including angiotensin II and transforming growth factor-ß appeared to modulate expression of integrins in a paracrine or autocrine fashion. Our study demonstrated that inflammatory cytokine TNF also regulates integrin expression. After MI, there is an immediate and rapid upregulation of TNF- in the myocardium as part of the immune injury response system.¹⁸ Our in vitro study demonstrated that integrin expression can be altered by TNF-. When myocytes were exposed to TNF-, there was a time-dependent downregulation of expression of integrin ß1D (the adult isoform, Figure 6). Integrin ß1 is present in 2 isoforms in the rat heart, ß1A (fetal isoform) and ß1D (adult isoform).²⁷ They are coexpressed in embryo heart until embryonic day 17. From this stage on, ß1A progressively decreased and ß1D increased to become the dominant ß-isoform in the adult cardiomyocytes.¹⁰ They share significant sequence homology in their alternatively spliced regions. Knockin of ß1 integrin with only the ß1D isoform was embryonically lethal because of the lack of proper heart formation, accompanied by multiple other defects,¹⁴ suggesting an essential function of ß1A to confer cell motility and plasticity during embryogenesis.¹⁴ ß1D, the dominant adult form of ß1 in the myocardium, is found in the intercalated disks at cell–cell contact points of the cardiomyocytes.¹⁹ ß1D interacts with structural proteins of the myocyte to form a stable cytoskeletal framework, permitting a stronger contractile force in the adult myocyte.²⁴ Adult ß1D knockout mice display abnormalities of cardiac function. Our study suggests that transition of the integrin from the ß1D to ß1A isoform in the myocardium after infarction may be regulated by cytokines such as TNF and may contribute to the decreased function and increased mobility of the myocytes to facilitate the remodeling process.

Another important development of the wound-healing process and inflammation is reestablishment of the capillary network, or angiogenesis. Angiogenesis is characterized by the invasion, migration, and proliferation of smooth muscle and endothelial cells. Several members of the integrin family of adhesion receptors are expressed on the surface of cultured smooth muscle cells and endothelial cells.^29–31 We found integrin ß3 mRNA and protein expressed primarily at the edge of infarcts, associated with vascular structures (Figure 5). Recent studies demonstrated that integrin vß3 is strongly expressed in endothelial cells of new blood vessels in tumors⁷ and oxygen-induced retinal neovascularization.³² Endothelial cells migrate on extracellular matrix components in vitro and very likely also in vivo.³³ Cultured endothelial cells are dependent on vß3 for survival. If vß3 is blocked with arginine-glycine–aspartic acid motif (RGD)–containing peptides or antibodies selective for this integrin, endothelial cells grown in vitro and angiogenic endothelial cells in vivo will undergo apoptosis^7,16 and inhibit the migration of smooth muscle cells.¹⁵ These observations suggest that integrin ß3 upregulation may contribute to angiogenesis in the peri-infarct zone as part of the remodeling process.

In summary, we studied temporal and spatial responses of integrin ß1 and ß3 in the rat heart after MI. Integrin ß1 and ß3 expression is upregulated at the site of MI in both inflammatory and fibrogenic stages of healing. Integrin ß subunits may play an important role in tissue repair and angiogenesis. The expressions of integrins in cardiac cells are also regulated by TNF-, which can uniquely induce isoform transition from ß1D to ß1A dominance. This may account for some of the important effects of TNF on cardiac remodeling after MI.

	Acknowledgments

 
This study was supported in part by grants from the Heart and Stroke Foundation of Ontario and the Canadian Institutes of Health Research. Dr Liu is the Heart and Stroke/Polo Chair Professor of Medicine at the University of Toronto.

	Footnotes

 
A supplemental Methods section, including additional figures, is available in the online-only Data Supplement at http://www.circulationaha.org.

Received August 9, 2002; revision received November 7, 2002; accepted November 8, 2002.

	References

Top Abstract Introduction Methods Results Discussion References

 

1. Hemler ME. VLA protein in the integrin family: structure, functions and their role on leukocytes. Annu Rev Immunol. 1990; 8: 365–400.[CrossRef][Medline] [Order article via Infotrieve]
   
2. Hynes RO. Integrins: versatility, modulation and signaling in cell adhesion. Cell. 1992; 69: 11–25.[CrossRef][Medline] [Order article via Infotrieve]
   
3. Meredith JEJ, Winitz S, Lewis JM, et al. The regulation of growth and intracellular signaling by integrins. Endocr Rev. 1996; 17: 207–220.[CrossRef][Medline] [Order article via Infotrieve]
   
4. Schwartz MA, Challer MD, Ginsberg MH. Integrins. Emerging paradigms of signal transduction. Annu Rev Cell Biol. 1995; 11: 543–599.
   
5. Cass DL, Bullard KM, Sylvester KG, et al. Epidermal integrin expression is upregulated rapidly in human fetal wound repair. J Pediatr Surg. 1998; 33: 312–316.[CrossRef][Medline] [Order article via Infotrieve]
   
6. Harwood FL, Monosov AZ, Goomer RS, et al. Integrin expression is upregulated during early healing in a canine intrasynovial flexor tendon repair and controlled passive motion model. Connect Tissue Res. 1998; 39: 309–316.[Medline] [Order article via Infotrieve]
   
7. Brooks PC, Montgomery AM, Rosenfeld M, et al. Integrin vß3 antagonists promote tumor regression by inducing apoptosis of angiogenic blood vessels. Cell. 1994; 79: 1157–1164.[CrossRef][Medline] [Order article via Infotrieve]
   
8. Ding B, Price RL, Goldsmith EC, et al. Left ventricular hypertrophy in ascending aortic stenosis mice: anoikis and the progression to early failure. Circulation. 2000; 101: 2854–2862.[Abstract/Free Full Text]
   
9. Nawata J, Ohno I, Isoyama S, et al. Differential expression of 1, 3 and 5 integrin subunits in acute and chronic stages of myocardial infarction in rats. Cardiovasc Res. 1999; 43: 371–381.[Abstract/Free Full Text]
   
10. Brancaccio M, Cabodi S, Belkin AM, et al. Differential onset of expression of 7 and ß1D integrins during mouse heart and skeletal muscle development. Cell Adhes Commun. 1998; 15: 193–205.
    
11. Zhidkova NI, Belkin AM, Maynel R. Novel isoform of ß1 integrin expressed in skeletal and cardiac muscle. Biochem Biophys Res Commun. 1995; 214: 279–285.[CrossRef][Medline] [Order article via Infotrieve]
    
12. Ingber DE. Cellular basis of mechanotransduction. Biol Bull. 1998; 194: 323–325.[Medline] [Order article via Infotrieve]
    
13. Simpson DG, Majeski M, Borg TK, et al. Regulation of cardiac myocyte protein turnover and myofibrillar structure in vitro by specific directions of stretch. Circ Res. 1999; 85: e59–e69.[Medline] [Order article via Infotrieve]
    
14. Baudoin C, Goumans MJ, Mummery C, et al. Knockout and knockin of the ß1 exon D define distinct roles for integrin splice variants in heart function and embryonic development. Genes Dev. 1998; 12: 1202–1216.[Abstract/Free Full Text]
    
15. Slepian MJ, Massia SP, Dehdashti B, et al. ß3-Integrins rather than ß1-integrins dominate integrin-matrix interactions involved in postinjury smooth muscle cell migration. Circulation. 1998; 97: 1818–1827.[Abstract/Free Full Text]
    
16. Brooks PC, Stromblad S, Klemke R, et al. Antiintegrin vß3 blocks human breast growth and angiogenesis in human skin. J Clin Invest. 1995; 96: 1815–1822.[Medline] [Order article via Infotrieve]
    
17. Brooks PC, Clark RA, Cheresh DA. Requirement of vascular integrin vß3 for angiogenesis. Science. 1994; 264: 569–571.[Abstract/Free Full Text]
    
18. Irwin M, Mak S, Mann D, et al. Tissue expression and immunolocalization of tumour necrosis factor- in postinfarction dysfunctional myocardium. Circulation. 1999; 99: 1492–1498.[Abstract/Free Full Text]
    
19. Belkin A, Zidhkova N, Balzac F, et al. ß1D integrin displaces the ß1 isoform in striated muscle: localization at the junctional structures and signalling potential in nonmuscle cells. J Cell Biol. 1996; 132: 211–226.[Abstract/Free Full Text]
    
20. Komuro I. Molecular mechanism of mechanical stress-induced cardiac hypertrophy. Jpn Circ J. 2000; 41: 117–129.[CrossRef]
    
21. Heling A, Zimmermann R, Kostin S, et al. Increased expression of cytoskeletal, linkage, and extracellular proteins in failing human myocardium. Circ Res. 2000; 86: 846–853.[Abstract/Free Full Text]
    
22. Kuppuswamy D, Kerr C, Narishige T, et al. Association of tyrosine-phosphorylated c-src with the cytoskeleton of hypertrophying myocardium. J Biol Chem. 1997; 272: 4500–4508.[Abstract/Free Full Text]
    
23. Ross RS, Pham C, Shai SY, et al. Beta-1 integrins participate in the hypertrophic response of rat ventricular myocytes. Circ Res. 1998; 82: 1160–1172.[Abstract/Free Full Text]
    
24. Belkin AM, Retta SF, Pletjushkina OY, et al. Muscle beta-1D integrin reinforces the cytoskeleton-matrix link: modulation of integrin adhesive function by alternative splicing. J Cell Biology. 1997; 139: 1583–1595.[Abstract/Free Full Text]
    
25. Bissell DM. Hepatic fibrosis as wound repair: a progress report. J Gastroenterol. 1998; 33: 295–302.[CrossRef][Medline] [Order article via Infotrieve]
    
26. Levine D, Rockey DC, Milner TA, et al. Expression of the integrin 8ß1 during pulmonary and hepatic fibrosis. Am J Pathol. 2000; 156: 1927–1935.[Abstract/Free Full Text]
    
27. Sheppard D. Integrin-mediated activation of transforming growth factor-beta(1) in pulmonary fibrosis. Chest. 2001; 120: S49–S53.[Abstract/Free Full Text]
    
28. Hillis GS, Macleod AM. Integrins and disease. Clin Sci. 1996; 91: 639–650.[Medline] [Order article via Infotrieve]
    
29. Cheng YF, Kramer RH. Human microvascular endothelial cells express integrin-related complexes that mediate adhesion to the extracellular matrix. J Cell Physiol. 1989; 139: 2275–2286.
    
30. Cheresh DA. Human endothelial cells synthesize and express an Arg-Gly-Asp-directed adhesion receptor involved in attachment to fibrinogen and von Willebrand factor. Proc Natl Acad Sci U S A. 1987; 84: 6471–6475.[Abstract/Free Full Text]
    
31. Janat MF, Argraves WS, Liau G. Regulation of vascular smooth muscle cell integrin expression by transforming growth factor beta1 and by platelet-derived growth factor-BB. J Cell Physiol. 1992; 151: 588–595.[CrossRef][Medline] [Order article via Infotrieve]
    
32. Hammes HP, Brownlee M, Jonczyk A, et al. Subcutaneous injection of a cyclic peptide antagonist of vitronectin receptor-type integrins inhibits retinal neovascularization. Nat Med. 1996; 2: 529–533.[CrossRef][Medline] [Order article via Infotrieve]
    
33. Ruoslahti E, Engvall E. Integrins and vascular extracellular matrix assembly. J Clin Invest. 1997; 100: 53S–56S.
    

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				M. Brancaccio, E. Hirsch, A. Notte, G. Selvetella, G. Lembo, and G. Tarone Integrin signalling: The tug-of-war in heart hypertrophy Cardiovasc Res, June 1, 2006; 70(3): 422 - 433. [Abstract] [Full Text] [PDF]

				B. B. Ayach, M. Yoshimitsu, F. Dawood, M. Sun, S. Arab, M. Chen, K. Higuchi, C. Siatskas, P. Lee, H. Lim, et al. Stem cell factor receptor induces progenitor and natural killer cell-mediated cardiac survival and repair after myocardial infarction PNAS, February 14, 2006; 103(7): 2304 - 2309. [Abstract] [Full Text] [PDF]

				J. Hua, L. W. Dobrucki, M. M. Sadeghi, J. Zhang, B. N. Bourke, P. Cavaliere, J. Song, C. Chow, N. Jahanshad, N. van Royen, et al. Noninvasive Imaging of Angiogenesis With a 99mTc-Labeled Peptide Targeted at {alpha}v{beta}3 Integrin After Murine Hindlimb Ischemia Circulation, June 21, 2005; 111(24): 3255 - 3260. [Abstract] [Full Text] [PDF]

				V. Sarin, R. D Gaffin, G. A Meininger, and M. Muthuchamy Arginine-glycine-aspartic acid (RGD)-containing peptides inhibit the force production of mouse papillary muscle bundles via {alpha}5{beta}1 integrin J. Physiol., April 15, 2005; 564(2): 603 - 617. [Abstract] [Full Text] [PDF]

				K.-H. Lee, K.-H. Jung, S.-H. Song, D. H. Kim, B. C. Lee, H. J. Sung, Y.-M. Han, Y. S. Choe, D. Y. Chi, and B.-T. Kim Radiolabeled RGD Uptake and {alpha}v Integrin Expression Is Enhanced in Ischemic Murine Hindlimbs J. Nucl. Med., March 1, 2005; 46(3): 472 - 478. [Abstract] [Full Text] [PDF]

				J. N. Tsoporis, A. Marks, A. Haddad, F. Dawood, P. P. Liu, and T. G. Parker S100B Expression Modulates Left Ventricular Remodeling After Myocardial Infarction in Mice Circulation, February 8, 2005; 111(5): 598 - 606. [Abstract] [Full Text] [PDF]

				M. Sun, F. Dawood, W.-H. Wen, M. Chen, I. Dixon, L. A. Kirshenbaum, and P. P. Liu Excessive Tumor Necrosis Factor Activation After Infarction Contributes to Susceptibility of Myocardial Rupture and Left Ventricular Dysfunction Circulation, November 16, 2004; 110(20): 3221 - 3228. [Abstract] [Full Text] [PDF]

				R. Sandhu, K. Teichert-Kuliszewska, S. Nag, G. Proteau, M. J. Robb, A. I.M. Campbell, M. A. Kuliszewski, M. J.B. Kutryk, and D. J. Stewart Reciprocal regulation of angiopoietin-1 and angiopoietin-2 following myocardial infarction in the rat Cardiovasc Res, October 1, 2004; 64(1): 115 - 124. [Abstract] [Full Text] [PDF]

				R. Zohar, B. Zhu, P. Liu, J. Sodek, and C. A. McCulloch Increased cell death in osteopontin-deficient cardiac fibroblasts occurs by a caspase-3-independent pathway Am J Physiol Heart Circ Physiol, October 1, 2004; 287(4): H1730 - H1739. [Abstract] [Full Text] [PDF]

				M. Nian, P. Lee, N. Khaper, and P. Liu Inflammatory Cytokines and Postmyocardial Infarction Remodeling Circ. Res., June 25, 2004; 94(12): 1543 - 1553. [Abstract] [Full Text] [PDF]

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