June 2023
Volume 64, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2023
Estrogen Receptor Signaling and Keratoconus
Author Affiliations & Notes
  • Tarab Ajjan
    Cellular Biology and Anatomy, Augusta University, Augusta, Georgia, United States
  • Hannah Youngblood
    Cellular Biology and Anatomy, Augusta University, Augusta, Georgia, United States
    Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia, United States
  • Theresa Akoto
    Cellular Biology and Anatomy, Augusta University, Augusta, Georgia, United States
  • Jingwen Cai
    Cellular Biology and Anatomy, Augusta University, Augusta, Georgia, United States
  • Hongfang Yu
    Cellular Biology and Anatomy, Augusta University, Augusta, Georgia, United States
  • Jason Sun
    Cellular Biology and Anatomy, Augusta University, Augusta, Georgia, United States
  • Dimitrios Karamichos
    North Texas Eye Research Institute, University of North Texas Health Science Center, Fort Worth, Texas, United States
    Pharmaceutical Sciences, University of North Texas Health Science Center, Fort Worth, Texas, United States
  • Yutao Liu
    Cellular Biology and Anatomy, Augusta University, Augusta, Georgia, United States
    James & Jean Culver Vision Discovery Institute, Augusta University, Augusta, Georgia, United States
  • Footnotes
    Commercial Relationships   Tarab Ajjan None; Hannah Youngblood None; Theresa Akoto None; Jingwen Cai None; Hongfang Yu None; Jason Sun None; Dimitrios Karamichos None; Yutao Liu None
  • Footnotes
    Support  R01EY023242, R21EY028671, R21EY028671S, P30EY031631, R01EY028888
Investigative Ophthalmology & Visual Science June 2023, Vol.64, 1673. doi:
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      Tarab Ajjan, Hannah Youngblood, Theresa Akoto, Jingwen Cai, Hongfang Yu, Jason Sun, Dimitrios Karamichos, Yutao Liu; Estrogen Receptor Signaling and Keratoconus. Invest. Ophthalmol. Vis. Sci. 2023;64(8):1673.

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

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Abstract

Purpose : Recent genome-wide association studies (GWAS) have identified >100 genomic loci associated with keratoconus (KC) and related phenotypes such as central corneal thickness (CCT), corneal resistance factor (CRF), and corneal hysteresis (CH). We aimed to prioritize these loci and identify the underlying pathways.

Methods : We collected the genes associated with CCT, CH, CRF, and KC from GWAS and prioritized genes based on their expression changes in KC-affected human corneas and corneal stromal cells using RNA-Seq. We performed pathway analyses using Ingenuity Pathway Analysis (IPA). We then examined the expression of estrogen receptors (ESR1, ESR2, and GPER1) in a known UCSC corneal single cell RNA-Seq (scRNA-Seq) dataset. Using RNA-Seq, we examined their expression in primary human corneal fibroblasts consisting of normal human corneal (n=4) and KC fibroblasts (n=4) treated with 0, 5, and 10ng/ml of TGFβ1 with and without 15% cyclic mechanical stretch (CMS) (1 cycle/s, 24 hours) followed by total RNA-Seq (n=48). A multi-factorial ANOVA model including KC status, TGFβ1 treatment, and CMS status was used to identify genes responsive to KC, TGFβ1, or CMS. We used droplet digital PCR (ddPCR) to validate expression of ESR1 and GPER1 in reference to GAPDH and HPRT1 genes. We examined the corneas of Esr1 and Gper1 knockout mice for potential corneal changes using a Bioptigen Envisu-R2200 OCT system (n=15-20 per genotype).

Results : Out of a total of 288 genes associated with KC, CH, CCT, and CRF, 119 genes passed filtering criteria based on their corneal expression. Our bioinformatics analysis identified gene networks centered on estrogen receptors and identified estrogen as an upstream regulator of our genes of interest, suggesting a potential role for estrogen signaling in KC. scRNA-Seq data indicated the expression of ESR1/GPER1 in human corneal epithelial and stromal cells. Our RNA-Seq data evidenced the expression of ESR1 and GPER1, but not ESR2, in human corneal fibroblasts. The ANOVA analysis found that GPER1 expression was induced 1.5-fold by CMS (FDR =0.005), and ESR1 expression was positively correlated with TGFβ1 treatment (FDR = 0.0002, r=0.61). Our ddPCR data indicated the same trend of expression changes. However, we did not observe significant changes in CCT in either Esr1 or Gper1 knockout mice.

Conclusions : Our in silico and experimental data revealed a potential role of estrogen receptor signaling in KC, consistent with epidemiology data.

This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.

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