The Charles T. Campbell Eye Microbiology Lab
UPMCUniversity of Pittsburgh Schools of the Health Sciences
HomeContact InformationLab Diagnostic TestingAntibiotic SusceptibilityAntimicrobial TherapyCurrent ResearchPhotos


Ocular Microbiology and Immunology Group
Back to OMIG Main Page

< Previous | 2020 Agenda and Abstracts | Next >

 

2020 OMIG Abstract

Proinflammatory Cytokines Induce Inflammasome-mediated Corneal Endothelial Cell Death
Andres Serrano, MD1,2, Angela Gomez1,2, Enrique Salero, PhD1,2, Arianna Tovar, MD1,2,
Robert W. Keane3, Juan Pablo de Rivero Vaccari3, Guillermo Amescua, MD1,
Alfonso L. Sabater, MD, PhD1,2
1Department of Ophthalmology, Bascom Palmer Eye Institute; 2Cornea and Ocular Surface Regenerative Medicine Center; 3Department of Neurological Surgery, Neuroscience Program, Miami Project to Cure Paralysis, University of Miami Miller School of Medicine, Miami, FL, USA

Purpose: Chronic corneal endothelial cell loss is one of the most common side effects of complicated intraocular surgery, resulting in corneal edema and vision loss in more than 2 million individuals a year. Low endothelial cell density has been associated with an elevation of intraocular pro-inflammatory cytokine levels. The inflammasome contributes to the activation of caspase-1 and the processing of the pro-inflammatory cytokine interleukin (IL)-1b as well as the cell death mechanism of pyroptosis through the cleavage of gasdermin-D (GSDM-D). The purpose of this study was to determine the role of the inflammasome in chronic corneal endothelial cell loss.

Methods:: Endothelial grafts from human donor corneas were treated ex vivo with the pro-inflammatory cytokines (tumor necrosis factor (TNF)-α and interferon (IFN)-γ) for 48 hours. The levels of caspase-1 and IL-1b were determined by ELISA. The expression of caspase-1 and GSDM-D were confirmed by immunofluorescence. Endothelial cell damage was determined by LDH release assay. Oxidative stress was measured by reactive oxygen species (ROS) assay. A specific caspase-1 inhibitor (Ac-YVAD-cmk) was then used to assess the involvement of caspase-1 in corneal endothelial cell death.

Results: Protein levels of caspase-1 and IL-1b were elevated in endothelial grafts treated with TNF-α and INF-γ. These findings where consistent with a higher intracellular expression of caspase-1 and GSDM-D. The levels of ROS were upregulated by TNF-α and INF-γ; whereas caspase-1, LDH released and ROS levels were all significantly downregulated by caspase-1 inhibition with Ac-YVAD-cmk.

Conclusion: Corneal endothelial cell death is induced by pro-inflammatory cytokines, TNF-α and INF-γ, and is mediated by caspase-1 inflammasome activation. Caspase-1 inhibitor (Ac-YVAD-cmk) may be an important drug target for chronic corneal endothelial cell loss.

Disclosure: RWK and JPdRV have licensed patents on the inflammasome (P). RWK and JPdRV are Scientific Advisory Board members of ZyVersa Thearapeutics (C).

Support: This research was supported by the Florida Lions Eye Bank, the Beauty of Sight Foundation (Dr. Sabater), NIH Center Core Grant P30EY014801 and Research to Prevent Blindness Unrestricted Grant.

 

 

< Previous | 2020 Agenda and Abstracts | Next >

 

 

 

space