June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
UVB activates autophagy in corneal epithelial stem cells
Author Affiliations & Notes
  • Thomas Volatier
    Department of Experimental Ophtalmology, Klinikum der Universitat zu Koln Zentrum fur Augenheilkunde, Koln, Nordrhein-Westfalen, Germany
  • Andreas Kluth
    TICEBA Gmbh, Germany
    Rheacell, Germany
  • Christoph Maßlo
    TICEBA Gmbh, Germany
    Rheacell, Germany
  • Christoph Ganss
    TICEBA Gmbh, Germany
    Rheacell, Germany
  • Markus Hermann Frank
    Harvard University, Cambridge, Massachusetts, United States
  • Natasha Frank
    Harvard University, Cambridge, Massachusetts, United States
  • Björn Schumacher
    9. Institute for Genome Stability in Ageing and Disease, Exzellenzcluster CECAD in der Universitat zu Koln, Koln, Nordrhein-Westfalen, Germany
  • Maria Notara
    Department of Experimental Ophtalmology, Klinikum der Universitat zu Koln Zentrum fur Augenheilkunde, Koln, Nordrhein-Westfalen, Germany
    University of Cologne Center for Molecular Medicine Cologne, Cologne, Nordrhein-Westfalen, Germany
  • Claus Cursiefen
    Department of Experimental Ophtalmology, Klinikum der Universitat zu Koln Zentrum fur Augenheilkunde, Koln, Nordrhein-Westfalen, Germany
    9. Institute for Genome Stability in Ageing and Disease, Exzellenzcluster CECAD in der Universitat zu Koln, Koln, Nordrhein-Westfalen, Germany
  • Footnotes
    Commercial Relationships   Thomas Volatier None; Andreas Kluth None; Christoph Maßlo None; Christoph Ganss None; Markus Frank None; Natasha Frank None; Björn Schumacher None; Maria Notara None; Claus Cursiefen None
  • Footnotes
    Support  CMMC Grant A 09, DFG Grant NO1395/1-1
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 3265 – A0300. doi:
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    • Get Citation

      Thomas Volatier, Andreas Kluth, Christoph Maßlo, Christoph Ganss, Markus Hermann Frank, Natasha Frank, Björn Schumacher, Maria Notara, Claus Cursiefen; UVB activates autophagy in corneal epithelial stem cells. Invest. Ophthalmol. Vis. Sci. 2022;63(7):3265 – A0300.

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

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Abstract

Purpose : Ultraviolet B (UVB) irradiation may induce DNA lesions in all directly exposed tissues. In the human body, only two tissues are chronically exposed to UV: the skin and the cornea. The cornea lacks some of the phsycical barrier properties that the skin has, this makes the cornea particularly vulnearable damage. The induction of autophagy, a process that removes damaged cytoplasmic components, is of great importance in the cornea and extremely so in the stem cells that maintain the cornea. Here, we aim to characterise autophagy in isolated corneal epithelial stem cells (CESC) exposed to UV.

Methods : Epithelial cells were extracted from the limbus of discarded donor cornea (n=3). The cells were sorted to remove any fibroblasts. Cells were irradiated with 0.05J/cm2 UVB. Autophagy and stem cells markers were observed using qPCR and immunofluorescent imaging. UVB damage was confirmed using DNA-lesion immunofluorescence assay.

Results : The putative stem cell character was assessed with ABCB5 and p63a. In culture, p63a+ epithelial cells have a high expression of mTOR that decreases by -0.96±0.6 log2 fold with UVB irradiation. Additionally, phosphrylated-mTOR (p-mTOR) position changes in CESC after irradiation, concentrating at the nucleus. Other assayed autophagy controllers had increased expression following UVB irradiation. ATG5 expression increased by 6.9±0.08 log2 fold, ATG7 expression increased by 7.7±2.9 log2 fold, ATG12 expression increased by 2.9±0.4 log2 fold, and LC3B expression increased by 1.05±0.3 log2 fold.

Conclusions : The decrease in expression of mTOR expression as well as the relocalisation of p-mTOR to the nucleus suggests that autophagy in CESC cytolasm is no longer inhibited. Additionally, both higher expression and visible presence of key autophagy controllers ATG5, ATG7, ATG12, and LC3B strongly suggests that short UVB exposure triggers autophagy in CESC.

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

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