June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
An in-depth protein analysis of scleritis using mass spectrometry
Author Affiliations & Notes
  • Daphne Vergouwen
    Ophthalmology, Erasmus MC, Rotterdam, Zuid-Holland, Netherlands
    Immunology, Erasmus MC, Rotterdam, Zuid-Holland, Netherlands
  • J.C. Ten Berge
    Ophthalmology, Erasmus MC, Rotterdam, Zuid-Holland, Netherlands
  • C. Guzel
    Neurology, Erasmus MC, Rotterdam, Zuid-Holland, Netherlands
  • Thierry van den Bosch
    Pathology, section Ophthalmic Pathology, Erasmus MC, Rotterdam, Zuid-Holland, Netherlands
  • Robert Verdijk
    Pathology, section Ophthalmic Pathology, Erasmus MC, Rotterdam, Zuid-Holland, Netherlands
  • A. Rothova
    Ophthalmology, Erasmus MC, Rotterdam, Zuid-Holland, Netherlands
  • Theo Luider
    Neurology, Erasmus MC, Rotterdam, Zuid-Holland, Netherlands
  • Marco Schreurs
    Immunology, Erasmus MC, Rotterdam, Zuid-Holland, Netherlands
  • Footnotes
    Commercial Relationships   Daphne Vergouwen None; J.C. Ten Berge None; C. Guzel None; Thierry van den Bosch None; Robert Verdijk None; A. Rothova None; Theo Luider None; Marco Schreurs None
  • Footnotes
    Support  Stichting Lijf en Leven under Grant [number L&L/mon/19-007]
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 1185 – A0039. doi:
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    • Get Citation

      Daphne Vergouwen, J.C. Ten Berge, C. Guzel, Thierry van den Bosch, Robert Verdijk, A. Rothova, Theo Luider, Marco Schreurs; An in-depth protein analysis of scleritis using mass spectrometry. Invest. Ophthalmol. Vis. Sci. 2022;63(7):1185 – A0039.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Scleritis is a severe inflammatory ocular disorder with unknown pathogenesis. We investigated sclera affected by non-infectious scleritis as well as healthy sclera for differentially expressed proteins using a mass spectrometry-based proteomics approach.

Methods : We collected scleral samples of eyes enucleated due to severe scleritis (n=3), and control scleral tissues (n=5), all exenterated eyes for eyelid carcinomas without scleral invasion. Clinical data of patients were retrospectively gathered from medical files. Scleral samples were microscopically scraped from tissue slides, and loaded on the LC-MS mass spectrometer after trypsin digestion. In addition, all samples were stained for immuno-histopathological evaluation. Results were analyzed using the proteomics software Scaffold (Proteome Software), and considering the Benjamini-Hochberg method for multiple testing a P-value lower than 0,00016 was considered significant.

Results : Mass spectrometry identified 629 proteins within the healthy and diseased scleral tissues, whereof collagen type I was the most abundantly expressed protein. Collagen type II to XII were also present. Fillagrin-2 was found to be significantly upregulated (P=0.0001) in scleritis, a protein that plays a crucial role in epidermal barrier function. Elongation factor 2 and vinculin were other proteins upregulated in scleritis compared to healthy scleral tissue, however they were not significantly different in this small sample set.

Conclusions : Using this highly powerful and innovative technique, we found that fillagrin-2 was upregulated in scleral samples from patients with non-infectious scleritis. Further research, ideally including more scleritis cases, is needed to validate our findings, and identify the role of fillagrin-2 in the pathogenesis of scleritis.

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.

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