June 2020
Volume 61, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2020
Hyperosmolarity disrupts corneal barrier via TNF-α-induced Cathepsin S with suppressed anti-inflammatory cytokine IL-37 in primary human corneal epithelial cells
Author Affiliations & Notes
  • Jinmiao Li
    Ocular Surface Center, Cullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine, Houston, Texas, United States
    State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, Guangdong, China
  • Yun Zhang
    Ocular Surface Center, Cullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine, Houston, Texas, United States
    Zhejiang Eye Hospital, School of Optometry and Ophthalmology, Wenzhou Medical University, Hangzhou, Zhejiang, China
  • Stephen Pflugfelder
    Ocular Surface Center, Cullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine, Houston, Texas, United States
  • De-Quan Li
    Ocular Surface Center, Cullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine, Houston, Texas, United States
  • Footnotes
    Commercial Relationships   Jinmiao Li, None; Yun Zhang, None; Stephen Pflugfelder, None; De-Quan Li, None
  • Footnotes
    Support  NIH NEI Grants EY023598 (DQL) and EY011915 (SCP), Core Grant for Vision Research EY002520, Lions Foundation for Sight, Research to Prevent Blindness, Oshman Foundation, William Stamps Farish Fund.
Investigative Ophthalmology & Visual Science June 2020, Vol.61, 146. doi:
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      Jinmiao Li, Yun Zhang, Stephen Pflugfelder, De-Quan Li; Hyperosmolarity disrupts corneal barrier via TNF-α-induced Cathepsin S with suppressed anti-inflammatory cytokine IL-37 in primary human corneal epithelial cells. Invest. Ophthalmol. Vis. Sci. 2020;61(7):146.

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

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Abstract

Purpose : To explore the pathological role and molecular mechanism by which hyperosmolarity disrupts corneal epithelial barrier through TNF-α-induced protease cathepsin S with suppressed anti-inflammatory cytokine IL-37 using an in vitro dry eye model with human corneal epithelial cells (HCECs).

Methods : Primary HCECs were established from fresh donor limbal tissue explants. The cultures in iso-osmolar medium were switched to medium with increasing osmolarity (350, 400, 450 and 500 mOsM), without or with prior incubation of IL-37 for different time periods (2-48 hours). The integrity of apical barrier junction proteins was evaluated by immunostaining and confocal laser scanning microscopy. The expression of cytokines and cathepsin S by HCECs and the condition medium was determined by RT-qPCR, immunofluorescent staining and ELISA.

Results : The integrity of corneal epithelial barrier was observed to be largely disrupted as shown by 3D confocal laser scanning images of immunofluorescent staining for major junction proteins ZO-1, occludin, claudin 1, and E-cadherin, in HCECs exposed to 450-500 mOsM, when compared with iso-osmolar medium at 312 mOsM. The mRNA expression and protein production of pro-inflammatory cytokines (TNF-α, IL-1β and IL-6) were dramatically stimulated by hyperosmotic stress as evaluated by RT-qPCR and ELISA. The protease cathepsin S also increased significantly at both mRNA and protein levels, as evaluated by RT-qPCR and immunostaining. However, anti-inflammatory cytokine IL-37 was observed to decrease significantly as early as 4 hour and last 24 hours in HCECs exposed to hyperosmotic medium. Interestingly, recombinant human IL-37 was observed to suppress the production of pro-inflammatory cytokines and inhibit TNF-α-induced cathepsin S in HCECs exposed to hyperosmotic medium. Further investigations on molecular pathway are currently underway.

Conclusions : Our findings demonstrate that the hyperosmotic stress disrupts the corneal epithelial barrier through stimulating pro-inflammatory cytokines and protease cathepsin S while suppressing anti-inflammatory cytokine IL-37. The results may provide a novel insight on molecular pathogenesis and potential therapeutic targets for dry eye patients.

This is a 2020 ARVO Annual Meeting abstract.

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