June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Modeling blood-brain barrier phenotypes in glaucoma with human pluripotent stem cells.
Author Affiliations & Notes
  • Sailee Sham Lavekar
    Department of Biology, Indiana University Purdue University Indianapolis, Indianapolis, Indiana, United States
    Medical and Molecular Genetics, Stark Neurosciences Research Institute, Indiana University School of Medicine, Indianapolis, Indiana, United States
  • Jason Hughes
    Department of Anatomy, Cell Biology, and Physiology, Indiana University School of Medicine, Terre Haute, Terre Haute, Indiana, United States
  • Kang-Chieh Huang
    Department of Biology, Indiana University Purdue University Indianapolis, Indianapolis, Indiana, United States
    Medical and Molecular Genetics, Stark Neurosciences Research Institute, Indiana University School of Medicine, Indianapolis, Indiana, United States
  • Cátia Gomes
    Medical and Molecular Genetics, Stark Neurosciences Research Institute, Indiana University School of Medicine, Indianapolis, Indiana, United States
    Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, Illinois, United States
  • Scott Canfield
    Department of Anatomy, Cell Biology, and Physiology, Indiana University School of Medicine, Terre Haute, Terre Haute, Indiana, United States
  • Jason S Meyer
    Medical and Molecular Genetics, Stark Neurosciences Research Institute, Indiana University School of Medicine, Indianapolis, Indiana, United States
    Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, Illinois, United States
  • Footnotes
    Commercial Relationships   Sailee Lavekar None; Jason Hughes None; Kang-Chieh Huang None; Cátia Gomes None; Scott Canfield None; Jason Meyer Wisconsin Alumni Research Foundation, Code P (Patent)
  • Footnotes
    Support  NIH R01EY024984, NIH R01EY033022, NIH U24EY033269, BrightFocus Foundation G2020369
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 2437 – F0381. doi:
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      Sailee Sham Lavekar, Jason Hughes, Kang-Chieh Huang, Cátia Gomes, Scott Canfield, Jason S Meyer; Modeling blood-brain barrier phenotypes in glaucoma with human pluripotent stem cells.. Invest. Ophthalmol. Vis. Sci. 2022;63(7):2437 – F0381.

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

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Abstract

Purpose : Glaucoma is a neurodegenerative disease that affects the retina, resulting in the degeneration of retinal ganglion cells (RGCs) and subsequent loss of vision. While RGCs are the primary cell type affected in the disease state, previous studies have demonstrated that blood brain barrier (BBB) properties may be compromised in glaucoma. However, the factors responsible for BBB disruption have remained elusive and thus, the goal of the study was to explore how changes in paracrine signaling within the BBB may be responsible for certain aspects of glaucoma using human pluripotent stem cell (hPSC)-derived cells.

Methods : To model aspects of barrier dysfunction in glaucoma, hPSCs with a glaucoma-specific Optineurin (E50K) mutation, as well as isogenic controls, were directed to differentiate into cells of the BBB, including RGCs, astrocytes, and microvascular endothelial cells (MVECs). Subsequently, MVECs were seeded into the top of an established in vitro barrier transwell model, while RGCs and astrocytes grown in the bottom. The ability to recapitulate characteristic properties of the BBB were then tested, including trans-endothelial electrical resistance (TEER), permeability and rhodamine transport. Further, RNA sequencing and ELISA identified factors responsible for changes observed.

Results : The establishment of the barrier model with RGCs, astrocytes, and MVECs resulted in a significant enhancement of barrier properties compared to MVECs grown alone. However, when the barrier model was assembled with cells with the glaucoma OPTN(E50K) mutation, barrier integrity was compromised, including reduced TEER and increased barrier permeability along with reduced efflux transporter activity and/or localization. An increased expression of certain secreted factors were identified from OPTN(E50K) astrocytes, and addition of exogenous factors applied to healthy cultures resulted in barrier dysfunction similar to the glaucoma barrier model.

Conclusions : The results of this study will expand our understanding of the mechanisms by which glaucoma affects the BBB. Overall, the in vitro glaucoma barrier model demonstrated characteristic functional deficits, mimicking some aspects of the phenotypes observed during the progression of glaucomatous neurodegeneration, with strong implications for the use of this model for disease modeling and drug development to rescue neurodegenerative phenotypes.

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

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