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Patient-derived cornea organoids as drug repurposing models for aniridia-associated keratopathy
Ali Can Koc1, Gamze Kocak1, Burak Kahveci1
1İzmir Biomedicine and Genome Center, 35340, İzmir, Türkiye; İzmir International Biomedicine and Genome Institute, Dokuz Eylül University, 35340, İzmir, Türkiye.
Aims:
This study aims to investigate the efficacy of drug repurposing using a corneal organoid model developed from patient-derived iPSCs and to elucidate the pathophysiology of Aniridia-Associated Keratopathy (AAK).
Materials And Methods:
A 90-day stepwise differentiation protocol was used to generate corneal organoids from iPSC cell lines developed from aniridia patients and healthy control. The corneal organoids produced were characterized using histology, immunofluorescence, qPCR, western blot, and transcriptomics. Two known agents, Duloxetine and Ataluren, were tested for corneal organoids for the restoring PAX6 protein expression.
Key Findings:
Histological analyses showed that the corneal organoids had a similar architecture to the native corneal tissue. Corneal epithelial, stromal, and endothelial cell biomarker staining showed positive expressions. AAK corneal organoids exhibited features that indicate the AAK disease phenotype, such as thickening of the epithelial cell layers and decrease in expressions of PAX6, ΔNP63, and keratocan genes. An increase in PAX6 protein was observed in organoids produced from AAK1 after duloxetine treatment and in organoids produced from AAK2 following ataluren treatment. AAK3 did not respond to either agent, indicating that there was no mutation-specific drug activity. Transcriptomic analyses showed clear corneal differentiation and absence of retina or lens profile in organoids.
Significance:
This study presents patient-specific organoid models for AAK using iPSCs and offers insight into mutation effects and PAX6 restoration following drug repurposing. The findings form the basis of personalized treatments for congenital aniridia.
Insights
Patient-derived corneal organoids model Aniridia-Associated Keratopathy (AAK). Drug repurposing showed potential for restoring PAX6 protein expression, paving the way for personalized AAK treatments.
Area of Science:
- Ophthalmology
- Regenerative Medicine
- Genetics
Background:
- Aniridia-Associated Keratopathy (AAK) is a complex ocular condition.
- Understanding AAK pathophysiology is crucial for developing effective treatments.
- Patient-specific models are needed to study AAK and test therapeutic strategies.
Purpose of the Study:
- To investigate drug repurposing efficacy for AAK using patient-derived corneal organoids.
- To elucidate the pathophysiology of Aniridia-Associated Keratopathy (AAK).
- To establish patient-specific induced pluripotent stem cell (iPSC)-derived corneal organoid models for AAK research.
Main Methods:
- Generated corneal organoids from iPSC lines of aniridia patients and healthy controls over 90 days.
- Characterized organoids using histology, immunofluorescence, qPCR, western blot, and transcriptomics.
- Tested Duloxetine and Ataluren for their ability to restore PAX6 protein expression in AAK organoids.
Main Results:
- Corneal organoids exhibited native corneal tissue architecture and cell marker expression.
- AAK organoids showed disease phenotypes: epithelial thickening and decreased PAX6, ΔNP63, and keratocan gene expression.
- Duloxetine and Ataluren partially restored PAX6 expression in specific AAK organoid models, but AAK3 showed no response, indicating no mutation-specific drug activity.
Conclusions:
- Developed patient-specific iPSC-derived corneal organoid models for AAK.
- Gained insights into mutation effects and PAX6 restoration via drug repurposing.
- Findings support personalized treatment strategies for congenital aniridia.
