June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Differential innate immune response and replication of SARS-CoV-2 in human diabetic corneal epithelial cells
Author Affiliations & Notes
  • Sneha Singh
    Ophthalmology, Visual and Anatomical Sciences (OVAS), Wayne State University School of Medicine, Detroit, Michigan, United States
  • Gustavo Garcia
    University of California Los Angeles, Los Angeles, California, United States
  • Ruchi Shah
    Cedars-Sinai Medical Center, Los Angeles, California, United States
  • Andrei A Kramerov
    Cedars-Sinai Medical Center, Los Angeles, California, United States
  • Alokkumar Jha
    Stanford University School of Medicine, Stanford, California, United States
  • Vaithilingaraja Arumugaswami
    University of California Los Angeles, Los Angeles, California, United States
  • Alexander V Ljubimov
    Cedars-Sinai Medical Center, Los Angeles, California, United States
  • Ashok Kumar
    Ophthalmology, Visual and Anatomical Sciences (OVAS), Wayne State University School of Medicine, Detroit, Michigan, United States
  • Footnotes
    Commercial Relationships   Sneha Singh None; Gustavo Garcia None; Ruchi Shah None; Andrei Kramerov None; Alokkumar Jha None; Vaithilingaraja Arumugaswami None; Alexander Ljubimov None; Ashok Kumar None
  • Footnotes
    Support  R01EY026964, R01 EY027381, R21AI135583, RO1EY032149
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 1670 – A0500. doi:
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    • Get Citation

      Sneha Singh, Gustavo Garcia, Ruchi Shah, Andrei A Kramerov, Alokkumar Jha, Vaithilingaraja Arumugaswami, Alexander V Ljubimov, Ashok Kumar; Differential innate immune response and replication of SARS-CoV-2 in human diabetic corneal epithelial cells. Invest. Ophthalmol. Vis. Sci. 2022;63(7):1670 – A0500.

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

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Abstract

Purpose : Diabetes predisposes an individual to severe COVID-19. Diabetic cornea is also known to have impaired wound healing, increasing the chances of infection. Earlier, we reported the ability of SARS-CoV-2 to infect conjunctival cells, and the presence of viral RNA and proteins was also detected in the corneas of COVID-19 donors. In this study, we evaluated the effect of diabetes on corneal innate immune response during SARS-CoV-2 infection and sought to determine the underlying mechanisms.

Methods : Human primary corneolimbal epithelial cells (HCECs) were isolated from the corneas of three diabetic and three non-diabetic donors. In vitro studies were performed by infecting HCECs with SARS-CoV-2 – USA-WA1/2020 strain at MOI 0.5. Viral replication was assessed by viral genome copy number. RNAseq analysis was performed to determine genes/pathways altered by diabetic vs non-diabetic HCECs. qPCR was used to assess the expression of innate inflammatory and antiviral genes. Western blot was performed to detect the protein expression of antiviral signaling molecules.

Results : The primary HCECs were found permissive to SARS-CoV-2 infection, as evidenced by increased viral replication which peaked at day 3 p.i. along with an induction of p-STAT1. Interestingly, HCECs from diabetic cornea had higher viral RNA on all three days post-infection. SARS-CoV-2 infected HCECs exhibited induced expression of inflammatory genes and their levels were relatively higher in diabetic cells. RNA-seq analysis revealed significant differences in diabetic vs. non-diabetic SARS-CoV-2 infected cells with alteration in genes regulating viral response, inflammation, and injury. The most affected down-regulated genes are related to lipid metabolism, ferroptosis, and oxidative stress.

Conclusions : Our study demonstrates increased SARS-CoV-2 replication and differential innate antiviral and inflammatory response in HCECs from diabetic corneas. These results indicate that diabetes is a potential risk for enhanced infectivity of SARS-CoV-2 for the ocular surface.

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

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