Dataset Information

Upregulation of filaggrin 2 in scleritis using tissue mass spectrometry

ABSTRACT: Scleritis is a severe inflammatory ocular disorder with unknown pathogenesis. Using a mass spectrometry approach, we investigated healthy sclera and sclera affected by non-infectious scleritis for differentially expressed proteins. Therefore, we collected scleral samples of enucleated eyes due to severe non-infectious scleritis (n=3) and control scleral tissues (n=5), all exenterated eyes for eyelid carcinomas without scleral invasion. Nano LC-MS mass spectrometry identified 629 proteins within the healthy and diseased scleral tissues, of which collagen type I was the most abundantly expressed protein. Collagen type II-XII was also present. Filaggrin-2, a protein that plays a crucial role in epidermal barrier function, was upregulated in all scleritis cases. In addition, other epithelial-associated proteins were upregulated (such as Keratin 33b, 34, and 85, Epiplakin, transglutaminase-3, Galectin 7, and Caspase-14) in scleritis. Further, upregulated proteins involved in regulation of the cytoskeleton (Vinculin and Myosin 9) and housekeeping proteins were found (elongation factor-2 and cytoplasmic dynein 1). We selected two proteins for validation with immunohistochemistry: Fillagrin-2 and myosin 9. Upregulation of both proteins was confirmed, the latter protein showing co-localization with the endothelial cell marker ETC-related gene (ERG), indicating neovascularization in scleral tissue affected in scleritis. Further research, preferably with multiple scleritis cases, is needed to validate our findings, including identification of the specific role of Filaggrin-2 and angiogenesis in the pathogenesis of scleritis.

INSTRUMENT(S): Orbitrap Eclipse

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Coskun Guzel

LAB HEAD: Dr. T.M. Luider

PROVIDER: PXD038727 | Pride | 2023-05-10

REPOSITORIES: Pride

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Publications

Scleral Proteome in Noninfectious Scleritis Unravels Upregulation of Filaggrin-2 and Signs of Neovascularization.

Vergouwen Daphne P C DPC Ten Berge Josianne C JC Guzel Coskun C van den Bosch Thierry P P TPP Verdijk Robert M RM Rothova Aniki A Luider Theo M TM Schreurs Marco W J MWJ

Investigative ophthalmology & visual science 20230301 3

<h4>Purpose</h4>Scleritis is a severe inflammatory ocular disorder with unknown pathogenesis. We investigated healthy sclera as well as sclera affected by noninfectious scleritis for differentially expressed proteins using a mass spectrometry approach.<h4>Methods</h4>We collected scleral samples of enucleated eyes due to severe noninfectious scleritis (n = 3), and control scleral tissues (n = 5), all exenterated eyes for eyelid carcinomas (n = 4), or choroidal melanoma (n = 1) without scleral in ...[more]

PMID: 36930145

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Project description:Impaired drainage of aqueous humor through the trabecular meshwork (TM) culminating in increased intraocular pressure is a major risk factor for glaucoma, a leading cause of blindness worldwide. Regulation of aqueous humor drainage through the TM, however, is poorly understood. The role of RhoA GTPase-mediated contractile activity, cell adhesive interactions, and gene expression in regulation of aqueous humor outflow was investigated using adenoviral vector-driven expression of constitutively active RhoA (RhoAV14). Organ cultured anterior segments from porcine eyes expressing RhoAV14 exhibited significant reduction of aqueous humor outflow. Cultured TM cells expressing RhoAV14 revealed strong contractile cell morphology, increased actin stress fibers and focal adhesions, along with increased levels of phosphorylated myosin II, and collagen IV, fibronectin and laminin. cDNA microarray analysis of RNA extracted from RhoAV14 expressing human TM cells revealed a significant increase in the expression of genes encoding extracellular matrix (ECM) proteins, cytokines, integrins, cytoskeletal proteins and signaling proteins. Conversely, various ECM proteins stimulated robust increases in phosphorylation of myosin II, paxillin and focal adhesion kinase, and activated Rho GTPase and actin stress fiber formation in TM cells, indicating a potential regulatory feedback interaction between ECM-induced mechanical strain and Rho GTPase-induced isometric tension in TM cells. Collectively, these data demonstrate that sustained activation of Rho GTPase signaling in the aqueous humor outflow pathway increases resistance to aqueous humor outflow through the trabecular pathway by influencing the contractile force, cell adhesive interactions, and the expression of ECM proteins and cytokines in TM cells. Keywords: Gene Expression Two condition experiment: Human trabecular mesh work cells infected with Adenivirus expressing GFP Vs Adenovirus expressing GFP and constitutively active RhoAV14

Dataset Information

Upregulation of filaggrin 2 in scleritis using tissue mass spectrometry

Publications

Scleral Proteome in Noninfectious Scleritis Unravels Upregulation of Filaggrin-2 and Signs of Neovascularization.

Similar Datasets

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