An in vitro Model of Human Retinal Detachment Reveals Successive Death Pathway Activations

Jelena Potic; Martial Mbefo; Adeline Berger; Michael Nicolas; Dana Wanner; Corinne Kostic; Alexandre Matet; Francine Behar-Cohen; Alexandre Moulin; Yvan Arsenijevic

doi:10.3389/fnins.2020.571293

An in vitro Model of Human Retinal Detachment Reveals Successive Death Pathway Activations

Front Neurosci. 2020 Nov 26:14:571293. doi: 10.3389/fnins.2020.571293. eCollection 2020.

Authors

Jelena Potic^{1

2

3}, Martial Mbefo¹, Adeline Berger¹, Michael Nicolas¹, Dana Wanner¹, Corinne Kostic¹, Alexandre Matet^{1

4}, Francine Behar-Cohen^{1

5}, Alexandre Moulin¹, Yvan Arsenijevic¹

Affiliations

¹ Department of Ophthalmology, University of Lausanne, Jules-Gonin Eye Hospital, Fondation Asile des Aveugles, Lausanne, Switzerland.
² Clinic for Eye Diseases, Clinical Center of Serbia, Belgrade, Serbia.
³ Department of Ophthalmology, Faculty of Medicine, University of Belgrade, Belgrade, Serbia.
⁴ Department of Ophthalmology, Institut Curie, Université de Paris, Paris, France.
⁵ INSERM U 1138, Centre de Recherches des Cordeliers, Université Paris Descartes, Université Pierre et Marie Curie, Paris, France.

Abstract

Purpose: was to create an in vitro model of human retinal detachment (RD) to study the mechanisms of photoreceptor death.

Methods: Human retinas were obtained through eye globe donations for research purposes and cultivated as explants. Cell death was investigated in retinas with (control) and without retinal pigment epithelium (RPE) cells to mimic RD. Tissues were studied at different time points and immunohistological analyses for TUNEL, Cleaved caspase3, AIF, CDK4 and the epigenetic mark H3K27me3 were performed. Human and monkey eye globes with retinal detachment served as controls.

Results: The number of TUNEL-positive cells, compared between 1 and 7 days, increased with time in both retinas with RPE (from 1.2 ± 0.46 to 8 ± 0.89, n = 4) and without RPE (from 2.6 ± 0.73 to 16.3 ± 1.27, p < 0.014). In the group without RPE, cell death peaked at day 3 (p = 0.014) and was high until day 7. Almost no Cleaved-Caspase3 signal was observed, whereas a transient augmentation at day 3 of AIF-positive cells was observed to be about 10-fold in comparison to the control group (n = 2). Few CDK4-positive cells were found in both groups, but significantly more in the RD group at day 7 (1.8 ± 0.24 vs. 4.7 ± 0.58, p = 0.014). The H3K27me3 mark increased by 7-fold after 5 days in the RD group (p = 0.014) and slightly decreased at day 7 and was also observed to be markedly increased in human and monkey detached retina samples.

Conclusion: AIF expression coincides with the first peak of cell death, whereas the H3K27me3 mark increases during the cell death plateau, suggesting that photoreceptor death is induced by different successive pathways after RD. This in vitro model should permit the identification of neuroprotective drugs with clinical relevance.

Keywords: cell death pathways; human retina; in vitro model; photoreceptor death; retinal detachment.