Mast Cell Chymase Induces Smooth Muscle Cell Apoptosis by a Mechanism Involving Fibronectin Degradation and Disruption of Focal Adhesions

Leskinen, Markus; Lindstedt, Ken A.; Wang, Yenfeng; Kovanen, Petri T.

doi:10.1161/01.atv.0000051405.68811.4d

Cited by 109 publications

(96 citation statements)

References 53 publications

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“…This procedure allows identification of the initial state of cell activation before irreversible cell damage occurs, and further explores a more advanced state of cell injury characterized by nuclear and cytoplasmic abnormalities typical of apoptosis. The advantage of the proposed procedure is that early morphological changes (membrane blebbing and vesiculation) can be easily identified before anchorage-dependent survival of cells is compromised [19,32,[39][40][41][42]. In the absence of other morphological or structural modifications, these initial membrane changes indicate a reversible response to stimulation by stressors and can be used as indicators of cell stimulation/suffering.…”

Section: Results-discussionmentioning

confidence: 99%

Plasmin on adherent cells: from microvesiculation to apoptosis

Doeuvre

Laurent

Goux

et al. 2010

Biochemical Journal

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SYNOPSISCell activation by stressors is characterised by a sequence of detectable phenotypic cell changes. A given stimulus, depending on its strength, induces modifications in the activity of membrane phospholipid transporters and calpains, which lead to phosphatidylserine exposure, membrane blebbing and the release of microparticles (nanoscale membrane vesicles). This vesiculation could be considered as a warning signal that may be followed, if the stimulus is maintained, by cell detachment-induced apoptosis. In this study, plasminogen incubated with adherent cells is converted into plasmin by constitutively expressed tPA or uPA. Plasmin formed on the cell membrane then induces a unique response characterized by membrane blebbing and vesiculation. Hitherto unknown for plasmin, these membrane changes are similar to those induced by thrombin on platelets. If plasmin formation persists, matrix proteins are then degraded, cells lose their attachments and enter the apoptotic process, characterized by DNA fragmentation and specific ultrastructural features. Since other proteolytic or inflammatory stimuli may evoke similar responses in different types of adherent cells, the proposed experimental procedure can be used to distinguish activated adherent cells from cells entering the apoptotic process. Such a distinction is crucial for evaluating effects of mediators, inhibitors and potential therapeutic agents.Key words: Plasminogen, membrane blebbing, microparticles, apoptosis, electron microscopy INTRODUCTIONCell response to a number of stressors and inflammatory mediators is key to maintenance of tissue homeostasis. The initial response of activated cells may evolve to apoptosis depending on the type and strength of stimuli [1]. Early manifestations of this cell activation process are the formation of membrane blebs and the shedding of nanoscale membrane fragments known and designated hereafter as microparticles (MPs, 0.1 to 1 µm) [2]. Since the first discovery of MPs in platelet-free plasma [3,4], the most well known cellular MPs are those of platelet, leukocyte, erythrocyte and endothelial cell origin found in circulating blood [5]. A number of studies have demonstrated that stimulation of these cells is followed by the characteristic features of cell activation: increased levels of cytoplasmic calcium associated to exposure of phosphatidylserine and activation of calpains (EC 3.4.22) [6]. The increase in intracellular calcium induces a disordered state in the concerted activity of membrane transporter proteins (flippases, floppases and scramblases) that maintain the membrane phospholipid asymmetry of quiescent cells [7,8]. As a result, procoagulant phosphatidylserine is translocated from the inner leaflet to the external leaflet of the membrane. The activated calpains cleave cytoskeleton filaments and thereby facilitate membrane blebbing and shedding of MPs. In addition to their procoagulant activity, MPs carry at their surface identity antigens that characterize their cellular origin. Since they also convey prot...

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Section: Results-discussionmentioning

confidence: 99%

Plasmin on adherent cells: from microvesiculation to apoptosis

Doeuvre

Laurent

Goux

et al. 2010

Biochemical Journal

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“…However, tryptase is involved in tissue fibrosis, and chymase induces apoptosis of cells with relatively low differentiation, including vascular smooth muscle cells, bronchial smooth muscle cells, and cardiomyocytes [16][17][18][19][20]. In other words, if we presume that there are undifferentiated cells in the fovea, then with regard to the action of these serine proteases, idiopathic epiretinal membranes may develop when tryptase activity is predominant, and idiopathic macular holes may develop when chymase activity is predominant.…”

Section: Discussionmentioning

confidence: 99%

Expression Sites of Neural Stem Cell-Related Genes in the Monkey Retina

Shibata¹,

Ikeda²

2015

J Stem Cell Res Ther

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Objectives: Based on the hypothesis that undifferentiated retinal stem cell (RSC)-like cells exist in the fovea (the light-stressed, concave, avascular center of the retina where light is focused), we investigated the expression sites of neural stem cell (NSC)-related genes in the monkey retina.Methods: Cynomolgus monkeys were euthanized, and both eyes were then enucleated. Each eye was hemisected near the limbus, and flat-mounted retina samples were then prepared. Using a stereomicroscope, 1-mm x 1-mm blocks of the retina at the fovea, mid-periphery, and extreme periphery were then excised. These samples were used for real-time polymerase chain reaction analysis of the NSC-related gene (nestin, PAX6, and SOX2) expression at each site. Results:Nestin expression was high in the fovea, with a lower expression in the mid-periphery and extreme periphery. No differences in PAX6 gene expression were found in the fovea, mid-periphery, and extreme periphery. SOX2 expression was highest in the extreme periphery, with decreased expression in the mid-periphery and fovea. Conclusions:Our finding that nestin expression was highest in the fovea suggests that foveal retinal cells may have more undifferentiated characteristics that are different from retinal cells at other sites.

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“…Furthermore, Lazaar et al (39) previously showed that chymase reduces epidermal growth factor-induced proliferation of SMCs. An alternative explanation for how mMCP-4 may control SMC responses would be that the fibronectin fragments generated in the presence of mMCP-4 could be proapoptotic for SMCs (33), thus possibly preventing an exaggerated SMC expansion in response to OVA treatment. However, numerous other potential targets for chymase have been described, many of which could directly or indirectly affect SMCs and AHR, e.g., substance P, vasoactive intestinal peptide, endothelin-1, angiotensin I, IL-13, and C3a (3).…”

Section: Discussionmentioning

confidence: 99%

“…Another potential substrate for mMCP-4 could be fibronectin. Fibronectin has previously been shown to be a substrate both for human chymase and for mMCP-4 (31, 32) and, considering that proteolytic fibronectin fragments can be proapoptotic for SMCs (33), it was relevant to examine whether fibronectin is differentially processed in lungs from WT and mMCP-4 Ϫ/Ϫ animals. As shown in Fig.…”

Section: Degradation Of Smc Mitogens and Fibronectin By Mmcp-4mentioning

confidence: 99%

Mouse Mast Cell Protease 4 Is the Major Chymase in Murine Airways and Has a Protective Role in Allergic Airway Inflammation

Waern

Jonasson

Hjoberg

et al. 2009

The Journal of Immunology

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It is widely established that mast cells (MCs) have a harmful role in asthma, for example by secreting various proinflammatory substances stored within their secretory granule. However, in this study, we show that one of the substances stored within MC granule, chymase, in fact has a protective role in allergic airway inflammation, indicating that MCs may possess both harmful and protective activities in connection with this type of disease. Wild-type (WT) mice and mice lacking mouse MC protease 4 (mMCP-4), a chymase that is functionally homologous to human chymase, were sensitized and challenged with OVA, followed by the assessment of airway physiology and inflammatory parameters. Our results show that the airway hyperresponsiveness was significantly higher in mMCP-4−/− as compared with WT mice. Moreover, the degree of lung tissue inflammation was markedly higher in mice lacking mMCP-4 than in WT controls. Histological analysis revealed that OVA sensitization/challenge resulted in a marked increased in the thickness of the smooth muscle cell (SMC) layer and, notably, that the degree of SMC layer thickening was more pronounced in mMCP-4−/− animals than in WT controls, thus indicating that chymase may have an effect on airway SMCs. In support of this, mMCP-4-positive MCs were located in the close vicinity of the SMC layer, mainly in the upper airways, and mMCP-4 was shown to be the major chymase expressed in these MCs. Taken together, our results indicate that chymase present in the upper airways protects against allergic airway responses, possibly by regulating SMCs.

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Mast Cell Chymase Induces Smooth Muscle Cell Apoptosis by a Mechanism Involving Fibronectin Degradation and Disruption of Focal Adhesions

Abstract: Objective-Chymase released from activated mast cells has been shown to induce apoptosis of vascular smooth muscle cells (SMCs) in vitro. The proteolytic activity of chymase is essential for the proapoptotic effect, but the mechanism of chymase-induced apoptosis has remained unknown.

Cited by 109 publications

References 53 publications

Plasmin on adherent cells: from microvesiculation to apoptosis

Plasmin on adherent cells: from microvesiculation to apoptosis

Expression Sites of Neural Stem Cell-Related Genes in the Monkey Retina

Mouse Mast Cell Protease 4 Is the Major Chymase in Murine Airways and Has a Protective Role in Allergic Airway Inflammation

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