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| Status |
Public on Feb 19, 2025 |
| Title |
Small molecule treatment alleviates photoreceptor cilia defects in LCA5-deficient human retinal organoids |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Bialleleic pathogenic variants in LCA5 cause one of the most severe forms of Leber congenital amaurosis, an early-onset retinal disease that results in severe visual impairment. Here, we report the use of gene editing to generate isogenic LCA5 knock-out (LCA5 KO) induced pluripotent stem cells (iPSC) and their differentiation to retinal organoids. The molecular and cellular phenotype of the LCA5 KO retinal organoids was studied in detail and compared to isogenic controls as well as patient-derived retinal organoids. The absence of LCA5 was confirmed in retinal organoids by immunohistochemistry and western blotting. There were no major changes in retinal organoid differentiation or ciliation, however, the localisation of CEP290 and IFT88 was significantly altered in LCA5 KO and patient photoreceptor cilia with extension along the axoneme. The LCA5-deficient organoids also had shorter outer segments and rhodopsin was mislocalised to the outer nuclear layer. We also identified transcriptomic and proteomic changes associated with the loss of LCA5. Importantly, treatment with the small molecules eupatilin, fasudil or a combination of both drugs improved rhodopsin traffic to the outer segment and reduced mislocalisation of rhodopsin in the outer nuclear layer. The treatments also effectively reduced CEP290 and IFT88 accumulation along the cilia. The improvements in cilia-associated protein localisation and traffic were accompanied by significant changes in the transcriptome towards control gene expression levels in many of the differentially expressed genes. In summary, iPSC-derived retinal organoids are a powerful model for investigating the molecular and cellular changes associated with loss of LCA5 function and highlight the therapeutic potential of small molecules to treat retinal ciliopathies.
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| Overall design |
Comparison of LCA5-knockout organoids treated with either eupatilin or fasudil against untreated knockout control organoids.
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| Contributor(s) |
Athanasiou D, Afanasyeva T, Chai N, Ziaka K, Jovanovic K, Guarascio R, Boldt K, Corral-Serrano J, Kanuga N, Roepman R, Collin R, Cheetham M |
| Citation(s) |
- Athanasiou D, Afanasyeva TAV, Chai N, Ziaka K et al. Small molecule treatment alleviates photoreceptor cilia defects in LCA5-deficient human retinal organoids. Acta Neuropathol Commun 2025 Feb 11;13(1):26. PMID: 39934925
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| Submission date |
Oct 10, 2024 |
| Last update date |
Feb 20, 2025 |
| Contact name |
Michael Cheetham |
| Organization name |
University College London
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| Department |
Institute of Ophthalmology
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| Street address |
11-43 Bath St
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| City |
London |
| ZIP/Postal code |
EC1V 9EL |
| Country |
United Kingdom |
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| Platforms (1) |
| GPL24676 |
Illumina NovaSeq 6000 (Homo sapiens) |
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