Wnt/frizzled signalling modulates the migration and differentiation of immortalized cardiac fibroblasts

Laeremans, Hilde; Rensen, Sander S.; Ottenheijm, H. C. J.; Smits, J. F. M.; Blankesteijn, W. Matthijs

doi:10.1093/cvr/cvq067

Cited by 58 publications

(54 citation statements)

References 47 publications

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“…However, some discrepancy exists, based on another report that failed to demonstrate increases in a-SMA expression upon the stimulation of fibroblasts with Wnt3a, despite increased migration (41). Moreover, another study showed decreased expression of smooth muscle a-actin after stimulation with Wnt3a (42). The causes of these disparate results are not clear, but seem to suggest that the functional outcome of the activation of the Wnt/b-catenin signaling pathway may be cell type-specific and context-specific.…”

Section: Discussionmentioning

confidence: 99%

Induction of a Mesenchymal Expression Program in Lung Epithelial Cells by Wingless Protein (Wnt)/β-Catenin Requires the Presence of c-Jun N-Terminal Kinase–1 (JNK1)

Velden

Guala

Leggett

et al. 2012

Am J Respir Cell Mol Biol

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Recent studies suggest the importance of the transition of airway epithelial cells (EMT) in pulmonary fibrosis, and also indicate a role for Wingless protein (Wnt)/b-catenin signaling in idiopathic pulmonary fibrosis. We investigated the possible role of the Wnt signaling pathway in inducing EMT in lung epithelial cells, and sought to unravel the role of c-Jun-N-terminal-kinase-1 (JNK1). The exposure of C10 lung epithelial cells or primary mouse tracheal epithelial cells (MTECs) to Wnt3a resulted in increases in JNK phosphorylation and nuclear b-catenin content. Because the role of b-catenin as a transcriptional coactivator is well established, we investigated T-cell factor/lymphocyteenhancement factor (TCF/LEF) transcriptional activity in C10 lung epithelial cells after the activation of Wnt. TCF/LEF transcriptional activity was enhanced after the activation of Wnt, and this increase in TCF/LEF transcriptional activity was diminished after the small interfering (si)RNA-mediated ablation of JNK. The activation of the Wnt pathway by Wnt3a, or the expression of either wild-type or constitutively active b-catenin (S37A), led to the activation of an EMT transcriptome, manifested by the increased mRNA expression of CArG box-binding factor-A, fibroblast-specific protein (FSP)-1, a-smooth muscle actin (a-SMA), and vimentin, increases in the content of a-SMA and FSP1, and the concomitant loss of zona occludens-1. The siRNA-mediated ablation of b-catenin substantially decreased Wnt3a-induced EMT. The siRNA ablation of JNK1 largely abolished Wnt3a, b-catenin, and b-catenin S37a-induced EMT. In MTECs lacking Jnk1, Wnt3a-induced increases in nuclear b-catenin, EMT transcriptome, and the content of a-SMA or FSP1 were substantially diminished. These data show that the activation of the Wnt signaling pathway is capable of inducing an EMT program in lung epithelial cells through b-catenin, and that this process is controlled by JNK1.Keywords: lung; epithelium; Wnt3a; fibrosis; epithelial to mesenchymal transitionThe development of fibrosis represents an important feature of pulmonary remodeling, and the critical role of epithelial cells in fibrogenesis is emerging. Studies from our laboratory identified a critical role for c-Jun-N-terminal-kinase-1 (JNK1) in augmenting the profibrotic effects of TGF-b1, in association with the causation of a mesenchymal transition of airway epithelialcells (EMT) (1). EMT is an important process during embryonic development, tumor progression, and fibrotic tissue repair after injury (2). We recently demonstrated that JNK1-induced phosphorylation in the linker domain of SMAD3 enhanced its ability to induce an EMT transcriptome. Consequently, JNK1-dependent, TGF-b1-induced EMT was greatly diminished in epithelial cells expressing a variant of SMAD3 refractory to phosphorylation in the linker domain (3).Recent studies indicated a role of b-catenin signaling in the induction of EMT. Moreover, the Wingless protein (Wnt)/ b-catenin axis was recently implicated to play a role in the development of idio...

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Section: Discussionmentioning

confidence: 99%

Induction of a Mesenchymal Expression Program in Lung Epithelial Cells by Wingless Protein (Wnt)/β-Catenin Requires the Presence of c-Jun N-Terminal Kinase–1 (JNK1)

Velden

Guala

Leggett

et al. 2012

Am J Respir Cell Mol Biol

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“…15,16 We tested seven different transfection reagents on cardiac fibroblasts with eGFP mRNA, and all of the reagents had very low transfection efficiency, except for lipofectamine 2000 (described henceforth as lipofectamine) which had a 20% transfection efficiency ( Figure S1). However, lipofectamine caused severe cell damage after 1 week of daily transfection, which is too short a period to induce direct cardiac reprogramming.…”

Section: Discussionmentioning

confidence: 99%

“…Traditional mRNA transfection methods have low efficiency and high toxicity for primary cardiac cells. 15,16 In addition, the toxicity of transfection reagents is a critical issue for cellular reprogramming because reprogramming requires multiple transfections over a period of weeks. 4,17 In this report, we present the demonstration of mRNAinduced cardiomyocyte-like gene expression in cardiac fibroblasts that were transfected with cardiac reprogramming factors in a new mRNA transfection system, termed C-Lipo, composed of a heart-targeting peptide (CRPPR-R9) and lipofectamine.…”

Section: Lee Et Almentioning

confidence: 99%

Peptide-enhanced mRNA transfection in cultured mouse cardiac fibroblasts and direct reprogramming towards cardiomyocyte-like cells

Lee¹,

Yu²,

Lingampalli³

et al. 2015

IJN

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The treatment of myocardial infarction is a major challenge in medicine due to the inability of heart tissue to regenerate. Direct reprogramming of endogenous cardiac fibroblasts into functional cardiomyocytes via the delivery of transcription factor mRNAs has the potential to regenerate cardiac tissue and to treat heart failure. Even though mRNA delivery to cardiac fibroblasts has the therapeutic potential, mRNA transfection in cardiac fibroblasts has been challenging. Herein, we develop an efficient mRNA transfection in cultured mouse cardiac fibroblasts via a polyarginine-fused heart-targeting peptide and lipofectamine complex, termed C-Lipo and demonstrate the partial direct reprogramming of cardiac fibroblasts towards cardiomyocyte cells. C-Lipo enabled the mRNA-induced direct cardiac reprogramming due to its efficient transfection with low toxicity, which allowed for multiple transfections of Gata4, Mef2c, and Tbx5 (GMT) mRNAs for a period of 2 weeks. The induced cardiomyocyte-like cells had α-MHC promoter-driven GFP expression and striated cardiac muscle structure from α-actinin immunohistochemistry. GMT mRNA transfection of cultured mouse cardiac fibroblasts via C-Lipo significantly increased expression of the cardiomyocyte marker genes, Actc1 , Actn2 , Gja1 , Hand2 , and Tnnt2 , after 2 weeks of transfection. Moreover, this study provides the first direct evidence that the stoichiometry of the GMT reprogramming factors influence the expression of cardiomyocyte marker genes. Our results demonstrate that mRNA delivery is a potential approach for cardiomyocyte generation.

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“…Other factors that contribute to myofibroblast differentiation include mechanical tension and ED-A fibronectin (Tomasek et al, 2002) and more recently, roles for Type VI collagen (Naugle et al, 2006) and Wnt/frizzled signaling (Laeremans et al, 2010) have been described in CF. Although not particularly well studied in the heart, there is evidence that TGF-induced differentiation of myofibroblasts from non-cardiac sources can be opposed by the action of proinflammatory cytokines, including IL-1 and TNF (Shephard et al, 2004;Liu et al, 2009).…”

Section: Myofibroblast Phenotypementioning

confidence: 99%

The role of cardiac fibroblasts in the transition from inflammation to fibrosis following myocardial infarction

Nieuwenhoven

Turner

2013

Vascular Pharmacology

123

108

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Cardiac fibroblasts (CF) play a pivotal role in the repair and remodeling of the heart that occurs following myocardial infarction (MI). The transition through the inflammatory, granulation and maturation phases of infarct healing is driven by cellular responses to local levels of cytokines, chemokines and growth factors that fluctuate in a temporal and spatial manner. In the acute inflammatory phase early after MI, CF contribute to the inflammatory milieu through increased secretion of proinflammatory cytokines and chemokines, and they promote extracellular matrix (ECM) degradation by increasing matrix metalloproteinase (MMP) expression and activity. In the granulation phase, CF migrate into the infarct zone, proliferate and produce MMPs and pro-angiogenic molecules to facilitate revascularization.Fibroblasts also undergo a phenotypic change to become myofibroblasts. In the maturation phase, inflammation is reduced by anti-inflammatory cytokines, and increased levels of profibrotic stimuli induce myofibroblasts to synthesize new ECM to form a scar. The scar is contracted through the mechanical force generated by myofibroblasts, preventing cardiac dilation. In this review we discuss the transition from myocardial inflammation to fibrosis with particular focus on how CF respond to alterations in proinflammatory and profibrotic signals. By furthering our understanding of these events, it is hoped that new therapeutic interventions will be developed that selectively reduce adverse myocardial remodeling post-MI, whilst sparing essential repair mechanisms.

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Wnt/frizzled signalling modulates the migration and differentiation of immortalized cardiac fibroblasts

Abstract: The results indicate that myofibroblast migration and differentiation, but not proliferation, can be modulated by interventions in Wnt/Fzd signalling. Therefore, Wnt/Fzd signalling may serve as a novel therapeutic target to ameliorate wound healing after MI.

Cited by 58 publications

References 47 publications

Induction of a Mesenchymal Expression Program in Lung Epithelial Cells by Wingless Protein (Wnt)/β-Catenin Requires the Presence of c-Jun N-Terminal Kinase–1 (JNK1)

Induction of a Mesenchymal Expression Program in Lung Epithelial Cells by Wingless Protein (Wnt)/β-Catenin Requires the Presence of c-Jun N-Terminal Kinase–1 (JNK1)

Peptide-enhanced mRNA transfection in cultured mouse cardiac fibroblasts and direct reprogramming towards cardiomyocyte-like cells

The role of cardiac fibroblasts in the transition from inflammation to fibrosis following myocardial infarction

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