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Heather L Chandler, Rachel B Matusow, Elizabeth Curto, Kristen J Gervais; Cyclosporine A Induces Autophagic Cell Death in Lens Epithelial Cells to Prevent Posterior Capsule Opacification Ex Vivo. Invest. Ophthalmol. Vis. Sci. 2014;55(13):3046.
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© ARVO (1962-2015); The Authors (2016-present)
To determine the appropriate Cyclosporine A (CsA) dose and minimum drug delivery time needed to prevent posterior capsule opacification (PCO) in an ex vivo model and evaluate the mechanism of CsA-induced cell death.
Lens capsules were harvested from canine cadaver eyes using an established ex vivo model of PCO. Lens capsules were treated with 0, 5, or 10 µg/mL CsA for 0, 2, 3, 4, 5, 6, or 7 days, and then maintained in culture for a total of 28 days in the absence of drug. CsA-treated lens epithelial cells (LEC) underwent routine transmission electron microscopy (TEM), western blotting, and fluorescent staining to evaluate the mechanism of cell death.
Lens capsules treated with 5, 6, or 7 days of 10 µg/mL CsA showed a significant decrease in ex vivo PCO formation; 7 days of drug delivery was sufficient to prevent PCO. Morphologically, CsA-treated LEC were swollen, had intact nuclei, lacked peripheral chromatin condensation, and demonstrated prominent vacuolization; TEM revealed autophagosomes. LC3-II protein expression and acridine orange fluorescence increased in CsA-treated cells. Dose dependent changes were observed in all experiments.
Seven days of intracapsular CsA drug delivery prevented ex vivo PCO formation. Morphologic changes and TEM suggest that CsA is able to induce LEC death via autophagy; this is a novel finding in the lens. Acridine orange is a marker for acidic vesicles and LC3-II is a protein involved in mediating autophagic death. Expression of these markers in CsA-treated LEC further supports this new mechanism of drug-induced LEC death.
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