June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Deletion of a cholesteryl ester hydrolase leads to a progressive retinal degeneration in mice
Author Affiliations & Notes
  • Seher Yuksel
    Ophthalmology, The University of Texas Southwestern Medical Center, Dallas, Texas, United States
  • Bogale Aredo
    Ophthalmology, The University of Texas Southwestern Medical Center, Dallas, Texas, United States
  • Yeshumenesh Zegeye
    Ophthalmology, The University of Texas Southwestern Medical Center, Dallas, Texas, United States
  • Cynthia X. Zhao
    Ophthalmology, The University of Texas Southwestern Medical Center, Dallas, Texas, United States
  • Miao Tang
    Center for the Genetics of Host Defense, The University of Texas Southwestern Medical Center, Dallas, Texas, United States
  • Xiaohong Li
    Center for the Genetics of Host Defense, The University of Texas Southwestern Medical Center, Dallas, Texas, United States
  • Laurent Gautron
    Department of Internal Medicine, Center for Hypothalamic Research, The University of Texas Southwestern Medical Center, Dallas, Texas, United States
  • Sara Ludwig
    Center for the Genetics of Host Defense, The University of Texas Southwestern Medical Center, Dallas, Texas, United States
  • Igor A Butovich
    Ophthalmology, The University of Texas Southwestern Medical Center, Dallas, Texas, United States
  • Bruce A. Beutler
    Center for the Genetics of Host Defense, The University of Texas Southwestern Medical Center, Dallas, Texas, United States
  • Rafael Ufret-Vincenty
    Ophthalmology, The University of Texas Southwestern Medical Center, Dallas, Texas, United States
  • Footnotes
    Commercial Relationships   Seher Yuksel None; Bogale Aredo None; Yeshumenesh Zegeye None; Cynthia Zhao None; Miao Tang None; Xiaohong Li None; Laurent Gautron None; Sara Ludwig None; Igor Butovich None; Bruce Beutler None; Rafael Ufret-Vincenty None
  • Footnotes
    Support  National Eye Institute grant-1R01EY033181, National Eye Institute grant-R01EY024324, VanSickle Family Foundation Grant, Pilot Synergy Grant, National Eye Institute Visual Science Core Grant EY020799, and David M. Crowley Foundation Grant
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 1952 – F0370. doi:
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      Seher Yuksel, Bogale Aredo, Yeshumenesh Zegeye, Cynthia X. Zhao, Miao Tang, Xiaohong Li, Laurent Gautron, Sara Ludwig, Igor A Butovich, Bruce A. Beutler, Rafael Ufret-Vincenty; Deletion of a cholesteryl ester hydrolase leads to a progressive retinal degeneration in mice. Invest. Ophthalmol. Vis. Sci. 2022;63(7):1952 – F0370.

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

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Abstract

Purpose : Lipid dyshomeostasis has a pathophysiologic role in multiple retinal diseases including age-related macular degeneration and diabetic retinopathy. Cholesteryl ester hydrolases are a family of enzymes found in many tissues, which are needed to convert the biologically inactive cholesteryl esters to their bioactive forms. Our goal was to characterize the role of a specific cholesteryl ester hydrolase (CEH), identified during a forward genetics screen, in retinal physiology.

Methods : We used a semiquantitative scale of fundus spots on retinal photos as a screening tool on a forward genetics pipeline (N-ethyl-N-nitrosourea mutagenesis of C57BL/6J mice). CRISPR/Cas9 technology was then used to generate knockout mice targeting the gene of interest. Fundus photos and Optical Coherence Tomography (OCT) were used to confirm the suspected phenotype. Further characterization of these mice was pursued using light microscopy, immunohistochemistry, in situ hybridization, electron microscopy, electrophysiology and retinal lipid analysis.

Results : We examined 5,793 G3 mice carrying 11,244 coding/splicing mutations in 7,679 genes. This corresponds to a genome saturation of 4.7% when considering predicted damaging mutations occurring in ≥2 mice in the homozygous state. Using the fundus spots scale we were able to identify a specific cholesteryl ester hydrolase as a gene of interest. A CRISPR/Cas9-generated KO mouse line targeting this CEH gene recapitulated the findings from the forward genetics screen. By 4 months of age, these CEH-deficient mice demonstrated accumulation of fundus spots (p< 0.001) compared to age and gender matched control mice. By 3 months of age, KO mice showed 7% thinning of the outer nuclear layer on OCT (p< 0.001) compared to control mice. These changes increased with aging. We will present anatomical, functional, ultrastructural and biochemical analyses of the retinas of these mice at different time points.

Conclusions : We demonstrate that a CEH enzyme, whose function has not been characterized in the retina before, is essential to retinal homeostasis.

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

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