WNT10B Enhances Proliferation through β-Catenin and RAC1 GTPase in Human Corneal Endothelial Cells

Lee, Jung Goo; Heur, Martin

doi:10.1074/jbc.m115.677245

Cited by 20 publications

(26 citation statements)

References 66 publications

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“…We have previously reported that cell migration and proliferation are independently regulated by WNT5A and WNT10B respectively in human corneal endothelial cells (43,44). In this study, we investigated the downstream targets of FGF2 that regulate cell proliferation and fibrosis in primary human CEC and human ex vivo corneal endothelium.…”

Section: Discussionmentioning

confidence: 97%

“…Protein Preparation, Protein Assay, SDS PAGE, and Immunoblotting Analysis -All assays were performed following previously reported protocols (12, 40,43,44). The following gel concentrations were used to separate proteins: 15% polyacrylamide gel for SNAI1 and SNAI2, 12% polyacrylamide gel for CDK2, 10% polyacrylamide gel for COL8A2, cyclin E1, vimentin, cytokeratin 12, αSMA, and β-actin, 8% polyacrylamide gel for E-cadherin, type I collagen, ZEB1 and ZEB2, and 6% polyacrylamide gel for Fibronectin.…”

Section: Methodsmentioning

confidence: 99%

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Fibroblast growth factor 2 induces proliferation and fibrosis via SNAI1-mediated activation of CDK2 and ZEB1 in corneal endothelium

Lee

Jung

Heur

2018

Journal of Biological Chemistry

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Investigating stimulation of endogenous wound healing in corneal endothelial cells (CECs) may help address the global shortage of donor corneas by decreasing the number of transplants performed for blindness due to endothelial dysfunction. We previously reported that IL-1β stimulation leads to fibroblast growth factor (FGF2) expression, enhancing migration and proliferation of mammalian CECs. However, FGF2 also promotes the endothelial-mesenchymal transition, which can lead to retrocorneal membrane formation and blindness. This prompted us to investigate downstream FGF2 signaling targets that could be manipulated to prevent retrocorneal membrane formation. FGF2 stimulation altered cell morphology and induced expression of mesenchymal transition marker genes such as snail family transcriptional repressor 1 (SNAI1), SNAI2, zinc finger E-boxbinding homeobox 1 (ZEB1), and ZEB2. This, in turn, induced expression of fibronectin, vimentin and, type I collagen and suppressed E-cadherin in CECs in vitro and ex vivo. siRNA-mediated SNAI1 knockdown revealed that SNAI1 induces ZEB1 expression, in turn inducing expression of type I collagen, the major component of retrocorneal membranes, and of cyclin-dependent kinase 2 (CDK2) and cyclin E1, promoting cell proliferation. siRNA-mediated knockdown of SNAI1 or ZEB1, but not of CDK2, inhibited FGF2-dependent expression of fibronectin, vimentin, and type I collagen and of suppression of E-cadherin expression. We conclude that SNAI1 is a key regulator of FGF2-dependent mesenchymal transition in human ex vivo corneal endothelium, with ZEB1 regulating type I collagen expression and CDK2 regulating cell proliferation. These results suggest that SNAI1 promotes fibrosis and cell proliferation in human corneal endothelium through ZEB1 and CDK2.The cornea is the anterior, transparent tissue of the human eye and consists of epithelium, stroma and endothelium. The corneal endothelium is composed of a monolayer of cells and plays a critical role in maintaining corneal transparency that is critical for sharp vision through its pump function (1,2). Adult human corneal endothelial cells (CEC) are usually mitotically inactive and arrested at the G 1 phase of the cell cycle (3,4). Because of the cell cycle arrest, there is a progressive decline in CEC density that can be further accelerated by injury, such as intraocular surgery or infection. When the density decreases below a critical threshold, corneal edema ensues, leading to loss of transparency and vision. Vision loss secondary to endothelial dysfunction is a common indication for corneal transplantation in developed nations.There have been many attempts to overcome the cell cycle arrest using various approaches including the use of growth factors such as FGF, EGF, and TGF-β, the use of endothelial cell growth supplements (5-9), and disruption of contact inhibition using EDTA (10). Moreover, severely injured CEC can undergo mesenchymal transition through which they lose their polarity and assume a fibroblastic phenotype. These CEC a...

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Section: Discussionmentioning

confidence: 97%

Section: Methodsmentioning

confidence: 99%

Fibroblast growth factor 2 induces proliferation and fibrosis via SNAI1-mediated activation of CDK2 and ZEB1 in corneal endothelium

Lee

Jung

Heur

2018

Journal of Biological Chemistry

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“…The Wnt signaling pathway is gaining prominence as an underappreciated player during inflammatory disorders such as atherosclerosis. In vitro, Wnt ligands have been shown to induce EC proliferation and modulate inflammation (24)(25)(26). Noncanonical Wnt5a/Ca 2+ -dependent signaling induces endothelial inflammation and release of inflammatory cytokines (27).…”

Section: Discussionmentioning

confidence: 99%

Endothelial cell–glucocorticoid receptor interactions and regulation of Wnt signaling

Zhou¹,

Mehta²,

Srivastava³

et al. 2020

JCI Insight

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“…Canonical Wnt proteins, namely, Wnt1, Wnt3a, Wnt7a, Wnt7b, and Wnt10b, have been demonstrated to activate ␤-catenin signaling in endothelial cells (ECs) and to regulate target genes that affect the angiogenic process (3)(4)(5)(6). In addition to its essential structural role in cadherin-based adhesions, ␤-catenin is a regulator of Wnt-mediated gene expression through its association in the nucleus with the T-cell factor/lymphoid-enhancing factor (TCF/LEF) (7)(8)(9).…”

mentioning

confidence: 99%

“…In the absence of Wnt stimulation, free cytoplasmic ␤-catenin levels are kept low by a degradation complex that includes glycogen synthase kinase 3␤ (GSK3␤), adenomatous polyposis coli (APC), and axin (10)(11)(12). Upon Wnt stimulation, ␤-catenin is stabilized in the cytoplasm and subsequently translocalized to the nucleus, interacts with TCF/LEF, and enhances the expression of genes involved in cell proliferation, differentiation, cell fate, and survival (6,(13)(14)(15).…”

mentioning

confidence: 99%

Endothelial NO Synthase-Dependent S-Nitrosylation of β-Catenin Prevents Its Association with TCF4 and Inhibits Proliferation of Endothelial Cells Stimulated by Wnt3a

Zhang

Chidiac

Delisle

et al. 2017

Molecular and Cellular Biology

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Nitric oxide (NO) produced by endothelial NO synthase (eNOS) modulates many functions in endothelial cells. S-nitrosylation (SNO) of cysteine residues on ␤-catenin by eNOS-derived NO has been shown to influence intercellular contacts between endothelial cells. However, the implication of SNO in the regulation of ␤-catenin transcriptional activity is ill defined. Here, we report that NO inhibits the transcriptional activity of ␤-catenin and endothelial cell proliferation induced by activation of Wnt/␤-catenin signaling. Interestingly, induction by Wnt3a of ␤-catenin target genes, such as the axin2 gene, is repressed in an eNOSdependent manner by vascular endothelial growth factor (VEGF). We identified Cys466 of ␤-catenin as a target for SNO by eNOS-derived NO and as the critical residue for the repressive effects of NO on ␤-catenin transcriptional activity. Furthermore, we observed that Cys466 of ␤-catenin, located at the binding interface of the ␤-catenin-TCF4 transcriptional complex, is essential for disruption of this complex by NO. Importantly, Cys466 of ␤-catenin is necessary for the inhibitory effects of NO on Wnt3a-stimulated proliferation of endothelial cells. Thus, our data define the mechanism responsible for the repressive effects of NO on the transcriptional activity of ␤-catenin and link eNOS-derived NO to the modulation by VEGF of Wnt/␤-catenin-induced endothelial cell proliferation.KEYWORDS VEGF, Wnts, cell signaling, endothelial cells, nitric oxide, nitric oxide synthase, S-nitrosylation T he Wnt/␤-catenin signaling pathway is involved in physiological and pathological angiogenesis, the process of blood vessel formation from preexisting vasculature, through its transcriptional regulation (1-3). Canonical Wnt proteins, namely, Wnt1, Wnt3a, Wnt7a, Wnt7b, and Wnt10b, have been demonstrated to activate ␤-catenin signaling in endothelial cells (ECs) and to regulate target genes that affect the angiogenic process (3-6). In addition to its essential structural role in cadherin-based adhesions, ␤-catenin is a regulator of Wnt-mediated gene expression through its association in the nucleus with the T-cell factor/lymphoid-enhancing factor (TCF/LEF) (7-9). In the absence of Wnt stimulation, free cytoplasmic ␤-catenin levels are kept low by a degradation complex that includes glycogen synthase kinase 3␤ (GSK3␤), adenomatous polyposis coli (APC), and axin (10-12). Upon Wnt stimulation, ␤-catenin is stabilized in the cytoplasm and subsequently translocalized to the nucleus, interacts with TCF/LEF, and enhances the expression of genes involved in cell proliferation, differentiation, cell fate, and survival (6, 13-15).In ECs, growth factors such as vascular endothelial growth factor (VEGF) regulate Citation Zhang Y, Chidiac R, Delisle C, Gratton J-P. 2017. Endothelial NO synthase-dependent S-nitrosylation of β-catenin prevents its association with TCF4 and inhibits proliferation of endothelial cells stimulated by Wnt3a. Mol

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WNT10B Enhances Proliferation through β-Catenin and RAC1 GTPase in Human Corneal Endothelial Cells

Cited by 20 publications

References 66 publications

Fibroblast growth factor 2 induces proliferation and fibrosis via SNAI1-mediated activation of CDK2 and ZEB1 in corneal endothelium

Fibroblast growth factor 2 induces proliferation and fibrosis via SNAI1-mediated activation of CDK2 and ZEB1 in corneal endothelium

Endothelial cell–glucocorticoid receptor interactions and regulation of Wnt signaling

Endothelial NO Synthase-Dependent S-Nitrosylation of β-Catenin Prevents Its Association with TCF4 and Inhibits Proliferation of Endothelial Cells Stimulated by Wnt3a

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