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Electroporation-Based Genetic Modification of Primary Human Pigment Epithelial Cells Using the Sleeping Beauty Transposon System

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Phototoxicity avoidance is a potential therapeutic approach for retinal dystrophy caused by EYS dysfunction.

Yuki Otsuka^1,2,3, Keiko Imamura^1,2,4, Akio Oishi⁵

¹iPSC-based Drug discovery and Development Team, RIKEN BioResource Research Center, Kyoto, Japan.

|April 22, 2024

Summary

Mutations in the eyes shut homolog (EYS) gene cause inherited retinal dystrophies (IRDs). EYS gene mutations disrupt photoreceptor cells, making them vulnerable to light, suggesting phototoxicity avoidance as a therapy for EYS-RD.

Keywords:

Ophthalmology Retinopathy Stem cells

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Subretinal Transplantation of Human Embryonic Stem Cell-Derived Retinal Tissue in a Feline Large Animal Model

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Published on: August 5, 2021

Area of Science:

Ophthalmology
Genetics
Cell Biology

Background:

Inherited retinal dystrophies (IRDs) cause progressive vision loss.
Mutations in the eyes shut homolog (EYS) gene are a frequent cause of IRDs.
The precise mechanism of photoreceptor degeneration in EYS-associated retinal dystrophy (EYS-RD) remains unclear.

Purpose of the Study:

To investigate the mechanism of photoreceptor cell degeneration in EYS-RD.
To utilize patient-derived induced pluripotent stem cells (iPSCs) to create retinal organoids for studying EYS-RD.
To explore the role of EYS and phototoxicity in the disease pathology.

Main Methods:

Generated retinal organoids from iPSCs of patients with EYS-RD.
Examined the localization of EYS and GRK7 in photoreceptor cells of the organoids.
Assessed the vulnerability of photoreceptor cells to light stimuli, particularly blue light.
Investigated the effect of EYS gene delivery on photoreceptor function.

Main Results:

Photoreceptor cells in EYS-RD organoids showed mislocalization of EYS and GRK7 from the outer segment.
These photoreceptor cells exhibited increased vulnerability to light, especially blue light.
Mislocalization of GRK7 was also observed in eys-knockout zebrafish models.
Restoring EYS in RD organoid photoreceptor cells corrected GRK7 mislocalization.

Conclusions:

EYS mutations lead to photoreceptor outer segment dysfunction and light sensitivity in EYS-RD.
Mislocalization of GRK7 contributes to phototoxicity in EYS-RD.
Targeting phototoxicity represents a potential therapeutic strategy for EYS-RD.