Aniridia – Novel therapeutic tools to treat or prevent progressive cornea opacification
Cell Biology
Final Report Abstract
The aim of this project was to develop a therapeutic strategy for patients with congenital aniridia who suffer from a progressive loss of corneal transparency, known as aniridiaassociated keratopathy. Congenital aniridia is a rare hereditary disease caused by mutation in the PAX6 gene, resulting in the inadequate production of PAX6 protein that is essential for the normal development and function of the eye. Currently, there is no effective treatment for this hereditary disease, which causes eye developmental disorders and severe visual loss from infancy to adulthood. In this project, three therapeutic candidates—duloxetine, ritanserin, and miRNA-204-5p mimic—were tested in various cell models to analyze their effects on PAX6 and other target gene expression. Materials and Methods (for the German project partner P3): Primary LECs were isolated from 5 aniridia patients and corneoscleral rims of 6 healthy human donors, that were transfected with small interfering RNA siRNA (PAX6 knockdown) and underwent 5 µM duloxetine and ritanserin treatment for 24 h. Using qPCR and western blot, gene and protein expression were analyzed. In addition, 5 primary limbal epithelial cell (LECs) cultures from healthy donors were transfected using Lipofectamine 2000 reagent and 100 pg/µl Has-miR-204-5p mimic RNA or the negative control (ctrl) miRNA. Using qPCR and western blot, gene and protein expression were analyzed. Results: In the siRNA based aniridia cell model and in primary aniridia limbal epithelial cells, expression of PAX6 and target genes, such as KRT12, KRT3, DSG1, ALDH1A1, ADH7, FABP5, ABCG2 did not change significantly at transcriptional or translational level, through duloxetine and ritanserin treatment (p≥0.4). In vitro, miR-204-5p overexpression suppressed vascular factor ANGPT1 in LECs at the gene and protein levels LEC. miR-204-5p mimics did not change PAX6 expression in LEC. Conclusions: Different culture model systems have various response to drug treatment and therefore need critical evaluation. The use of in vivo models can further advance our knowledge regarding duloxetine and ritanserin treatment in aniridia associated keratopathy in the future. While short-term miR-204-5p treatment effectively suppresses ANGPT1 in LECs, it does not affect PAX6 expression. Effects of longer-term treatment in in vivo models requires further study.
Publications
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Pathophysiology of aniridia-associated keratopathy: Developmental aspects and unanswered questions. The Ocular Surface, 22, 245-266.
Latta, L.; Figueiredo, F.C.; Ashery-Padan, R.; Collinson, J.M.; Daniels, J.; Ferrari, S.; Szentmáry, N.; Solá, S.; Shalom-Feuerstein, R.; Lako, M.; Xapelli, S.; Aberdam, D. & Lagali, N.
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miRNA Expression in primären Limbusepithelzellen von Aniridie Patienten. September 2022, Jahrestagung der Deutschen Ophthalmologischen Gesellschaft, Berlin, Deutschland
Nastaranpour N., Stachon T., Fries F.N., Ulrich M., Seitz B., Ludwig N., Meese E. & Szentmáry N.
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Pathophysiologie der Aniridie-assoziierten Keratopathie: Entwicklungsaspekte und unbeantwortete Fragen. 296. Freiburger Augenärzteabend 2022
Szentmáry N., Fries F.N., Stachon T., Ulrich M., Nastaranpour M., Latta L., Aberdam D., Lagali N., Seitz B. & Käsmann-Kellner B.
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Potential markers that discriminate between healthy and AAK patient cells. Nijmegen 2022
Szentmáry N., Fries F.N., Stachon T., Ulrich M., Nastaranpour M., Latta L., Aberdam D., Lagali N., Seitz B. & Käsmann-Kellner B.
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Aniridie-Keratopathie - Grundlagen und klinische Aspekte. Basler Fortbilungstage 2023, Eingeladenes Referat
Szentmáry N., Fries F.N., Náray A., Amini M., Suiwal S., Langenbucher A., Lagali N., Munteanu C., Corton-Perez M., Tory K., Csorba A., Nagy Z.Z., Csidey M., Maka E., Seitz B., Käsmann-Kellner B. & Stachon T.
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Corneal epithelial cell markers and microRNAs as markers of congenital aniridia. EVER Kongress, Valencia 2023
Szentmáry N., Nastaranpour M., Latta L., Ulrich M., Fries F.N., Aberdam D., Lagali N., Seitz B., Käsmann-Kellner B., Ludwig N. & Stachon T.
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Corneal epithelial cell markers in congenital aniridia. EVER Kongress, Valencia 2023
Szentmáry N., Nastaranpour M., Latta L., Ulrich M., Fries F.N., Aberdam D., Lagali N., Seitz B., Käsmann-Kellner B. & Stachon T.
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Gene expression in the siRNA based aniridia cell model and in primary aniridia limbal epithelial cells following duloxetine and ritanserin treatment. September 2023, Jahrestagung der Deutschen Ophthalmologischen Gesellschaft, Berlin, Deutschland
Suiwal S., Stachon T., Li Z., Nastaranpour M., Chai N., Amini M., Seitz B., Fries F.N., Tschernig T. & Szentmáry N.
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Identification of the regulatory circuit governing corneal epithelial fate determination and disease. PLOS Biology, 21(10), e3002336.
Smits, Jos G. A.; Cunha, Dulce Lima; Amini, Maryam; Bertolin, Marina; Laberthonnière, Camille; Qu, Jieqiong; Owen, Nicholas; Latta, Lorenz; Seitz, Berthold; Roux, Lauriane N.; Stachon, Tanja; Ferrari, Stefano; Moosajee, Mariya; Aberdam, Daniel; Szentmary, Nora; van Heeringen, Simon J. & Zhou, Huiqing
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MicroRNAs as markers of congenital aniridia. EVER Kongress, Valencia 2023
Stachon T., Amini M., Nastaranpour M., Suiwal S., Li Z., Chai N., Fries F.N., Seitz B., Ludwig N. & Szentmáry N.
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Effects of miR-204-5p modulation on PAX6 regulation and corneal inflammation. Scientific Reports, 14(1).
Abbasi, Mojdeh; Amini, Maryam; Moustardas, Petros; Gutsmiedl, Quirin; Javidjam, Dina; Suiwal, Shweta; Seitz, Berthold; Fries, Fabian N.; Dashti, Ava; Rautavaara, Yedizza; Stachon, Tanja; Szentmáry, Nóra & Lagali, Neil
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miRNA Expression Profile in Primary Limbal Epithelial Cells of Aniridia Patients. Investigative Ophthalmology & Visual Science, 66(1), 20.
Nastaranpour, Mahsa; Suiwal, Shweta; Stachon, Tanja; Fries, Fabian N.; Amini, Maryam; Seitz, Berthold; Meese, Eckart; Ludwig, Nicole & Szentmáry, Nóra
