Pterygial Derived Fibroblasts Express Functionally Active Histamine and Epidermal Growth Factor Receptors

doi:10.1006/exer.2001.1116

Experimental Eye Research

Volume 74, Issue 2, February 2002, Pages 237-244

https://doi.org/10.1006/exer.2001.1116 Get rights and content

Abstract

Pterygia are characterised by a fleshy outgrowth of altered conjuctival tissue over the cornea and are most common in tropical regions. Pterygial fibroblasts are characteristically distinct from normal conjunctival fibroblasts, and therefore the aim of this study was to determine the presence and functional significance of histamine and epidermal growth factor (EGF) receptors in these cells. Pterygial specimens were cultured in vitro and cellular outgrowths were phenotypically characterised as fibroblasts using vimentin and cytokeratin staining. Intracellular calcium mobilization was used to characterise the functional activity of histamine receptors on these cells. Maximal response was obtained with 100 μM histamine. However, lower concentrations of histamine also caused mobilization of calcium that were totally abolished by pre-incubation with H1 but not H2 or H3 receptor antagonists. EGF receptor was diffusely expressed over the cell surfaces. EGF stimulated receptor internalization, ERK protein phosphorylation and intracellular calcium mobilization. Therefore, fibroblasts derived from human pterygia express functionally active histamine and epidermal growth factor receptors. Controlled modification of either the receptors or the appropriate ligands could have beneficial effects in pterygia treatment.

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Proliferative factors related to pterygium pathogenesis have been noticed and reviewed many times in previous studies; therefore, these will be only briefly discussed here. These factors include heparin-binding EGF (HB-EGF), VEGF, PDGF, c-myc, secreted protein acidic and rich in cysteine (SPARC), urokinase-type plasminogen activator (uPA), and insulin-like growth factor binding protein-2 (IGFBP-2) (Kria et al., 1998; Shimmura et al., 2000; Maini et al., 2002; Jin et al., 2003; Solomon et al., 2003; Nolan et al., 2004; Cui et al., 2011; Seet et al., 2012; Chao et al., 2013) and, recently, diverse genes and proteins such as S100A8/9 are identified by microarrays and proteomic techniques (Hou et al., 2014). Eventually, these factors may be connected to diverse intracellular growth pathways and prolong proliferation of pterygium cells.
Activation of fibroblasts is a vital process during wound healing. However, if prolonged and exaggerated, profibrotic pathways lead to tissue fibrosis or scarring and further organ malfunction. Although the pathogenesis of pterygium is known to be multi-factorial, additional studies are needed to better understand the pathways initiated by fibroblast activation for the purpose of therapeutic translation. Regarding pterygium as a possible systemic disorder, we discuss the different cell types that pterygium fibroblasts originate from. These may include bone marrow-derived progenitor cells, cells undergoing epithelial–mesenchymal transition (EMT), and local resident stromal cells. We also describe how pterygium fibroblasts can be activated and perpetuate profibrotic signaling elicited by various proliferative drivers, immune-inflammation, and novel factors such as stromal cell-derived factor-1 (SDF-1) as well as a known key fibrotic factor, transforming growth factor-beta (TGF-β). Finally, epigenetic modification is discussed to explain inherited susceptibility to pterygium.
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Pterygium is a fibrovascular growth on the ocular surface with corneal tissue destruction, matrix degradation and varying extents of chronic inflammation. To facilitate investigation of pterygium etiology, we immortalized pterygium fibroblast cells and profiled their global transcript levels compared to primary cultured cells.
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Light microscopic morphology of immortalized cells was similar to primary pterygium fibroblast at passage 2–4. Telomerase reverse transcriptase was expressed, and p53 and p16 levels were reduced in immortalized pterygium fibroblast cells. There were 3308 significantly dysregulated genes showing at least 2 fold changes in transcript levels between immortalized and primary cultured cells (2005 genes were up-regulated and 1303 genes were down-regulated). Overall, 13.58% (95% CI: 13.08–14.10) of transcripts in immortalized cells were differentially expressed by at least 2 folds compared to primary cells.
Pterygium primary fibroblast cells were successfully immortalized to at least passage 11. Although a variety of genes are differentially expressed between immortalized and primary cells, only genes related to cell cycle are significantly changed, suggesting that the immortalized cells may be used as an in vitro model for pterygium pathology.
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Pterygium is a disease of the ocular surface that is associated with chronic UV exposure and is characterized by proliferation, inflammatory infiltrates, fibrosis, angiogenesis and extracellular matrix breakdown. Although pterygium is not fully understood, significant progress has been made toward understanding the mechanisms involved in its pathogenesis. In this review, we provide an update on the signaling pathways activated by UV light that result in induction of mediators responsible for the growth of pterygium. We also review the recent genetic studies on hereditary factors and provide a brief overview of the role of epithelial mesenchymal transition, bone marrow progenitor cells, and neuronal signals that may also contribute to the pathogenesis of pterygium. Therapeutic options for pterygium are discussed based on the mechanisms that perpetuate its growth.
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Over the past two decades, the applications of in vivo confocal microscopy to the investigation of ocular surface diseases in the living eye have been greatly extended. Confocal microscopy enables detailed investigation of tarsal and palpebral conjunctiva, central and peripheral cornea, tear film, and lids, and it allows evaluation of the ocular surface at the cellular level. High-quality imaging in both contact and noncontact modes has allowed new understanding of the functions of the ocular surface system, and in the coming years, such knowledge will become increasingly comprehensive and specific. Confocal microscopy may provide a link between well-established ex vivo histology and in vivo study of ocular pathology, not only in clinical science but also in clinical practice. The purpose of this review is to summarize the current knowledge about in vivo confocal microscopy of the ocular surface.
Epidermal growth factor receptor signaling is partially responsible for the increased matrix metalloproteinase-1 expression in ocular epithelial cells after UVB radiation
2005, American Journal of Pathology
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It would not be unreasonable to target cell-surface molecules such as the EGFR, as others have done in the setting of neoplasia.59 Not only is this receptor present in pterygia and pterygium-derived epithelial cells (Figure 8),33,34 it is phosphorylated by UVB, EGF, and HB-EGF, and is activated by viruses such as hepatitis B, Epstein-Barr, and Pox viruses.59 This, too, is relevant to the development of pterygia because there is on-going debate as to whether there is a viral component to this disease.60–62
Pterygia are inflammatory, invasive, and proliferative lesions of the human ocular surface in which the matrix metalloproteinase (MMP) collagenase-1 (MMP-1) is highly expressed. Pterygia development may involve MMP-1 activity against interstitial fibrillar collagen, an abundant extracellular matrix component of the cornea, and its induction by ultraviolet light (UVB). We examined the pathways responsible for enhanced expression of MMP-1 in pterygium epithelial cells after UVB exposure and/or treatment with chemical inhibitors of mitogen-activated protein kinases or epidermal growth factor receptor. The induction of MMP-1 by UVB was comparable to that mediated by heparin-binding epidermal growth factor-like growth factor and epidermal growth factor. The epidermal growth factor receptor inhibitor PD153035 partially blocked the UVB-mediated induction of MMP-1 and totally abrogated its production after stimulation with either heparin-binding epidermal growth factor-like growth factor or epidermal growth factor. UVB exposure enhanced the phosphorylated form of ERK1/2 in a time-dependent manner whereas the ERK1/2 inhibitor PD98059 decreased this induction by at least fivefold. Transcripts for c-jun and c-fos were detected as early as 2 hours after UVB exposure and were suppressed by PD98059. The identification of a specific intracellular signaling pathway responsible for the enhanced production of a key enzyme that denatures intact fibrillar collagen has important implications for understanding the pathophysiology and future therapy for pterygia.

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^f1: Address correspondence to: I. Michael Wormstone, School of Biological Sciences, University of East Anglia, Norwich, NR4 7TJ, U.K. E-mail: [email protected]

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Regular ArticlePterygial Derived Fibroblasts Express Functionally Active Histamine and Epidermal Growth Factor Receptors

Abstract

Rev. Fr. Allergol.

Exp. Eye Res.

Acta Histochem.

Exp. Eye Res.

Pharmacol. Ther.

Jpn. J. Ophthalmol.

Jpn. J. Ophthalmol.

Int. J. Radiat. Oncol.

Int. J. Radiat. Oncol.

Ophthalmology

Histology of pterygium: an electron microscopic study

Br. J. Ophthalmol.

The EGF receptor: a nexus for trafficking and signalling

Bioessays

The epidermal growth factor family

Handbook Exptl. Pharmacol.

Acute effects of in vitro mast-cell degranulation on human lung muscarinic receptors

Am. Rev. Respir. Dis.

Fibroblasts isolated from human pterygia exhibit transformed cell characteristics

In Vitro Cell Dev. Biol.

Cholecystokinin, carbachol, gastrin, histamine, and forskolin increase [Ca2+]i in gastric glands

Am. J. Physiol.

Pterygium as an early indicator of ultraviolet isolation: a hypothesis

Br. J. Ophthalmol.

Signal characteristics of G protein-transactivated EGF receptor

EMBO J.

P53 expression in altered limbal basal cells of pingueculae pterygia and limbal tumors

Curr. Eye Res.

A new generation of Ca2+ indicators with greatly improved fluorescence properties

J. Biol. Chem.

Pathogenesis of pterygium

Eye

Regular Article
Pterygial Derived Fibroblasts Express Functionally Active Histamine and Epidermal Growth Factor Receptors

Cholecystokinin, carbachol, gastrin, histamine, and forskolin increase [Ca²⁺]_i in gastric glands

A new generation of Ca²⁺ indicators with greatly improved fluorescence properties