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Erschienen in: Graefe's Archive for Clinical and Experimental Ophthalmology 10/2020

07.06.2020 | Genetics

Disease-causing mutations associated with bestrophinopathies promote apoptosis in retinal pigment epithelium cells

verfasst von: Tingting Gao, Chengqiang Tian, Hui Xu, Xin Tang, Lvzhen Huang, Mingwei Zhao

Erschienen in: Graefe's Archive for Clinical and Experimental Ophthalmology | Ausgabe 10/2020

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Abstract

Purpose

Best vitelliform macular dystrophy (BVMD) and autosomal recessive bestrophinopathy (ARB) are two kinds of bestrophinopathies which are caused by BEST1 mutations and characterized by accumulation of lipofuscin-like materials on the retinal pigment epithelium cell-photoreceptor interface. In the past two decades, research about the pathogenesis of bestrophinopathies was mainly focused on the anion channel and intracellular Ca2+ signaling, but seldom concentrated on the function of retinal pigment epithelium (RPE) cells. In this study, we explored the possible effect of the three BEST1 mutations p.V143F, p.S142G, and p.A146T on the apoptosis in human fetal RPE cells.

Methods

Wild-type plasmid and mutant plasmids BEST1-pcDNA3.1 p.V143F, p.S142G, and p.A146T were transfected to human fetal RPE cells. The molecules caspase-3, phospho-Bcl-2, BAX, PARP, and AIF associated with apoptosis were determined by quantitative PCR and Western blot. Apoptotic rate and active Caspase-3 staining were analyzed by flow cytometry.

Results

Caspase-3 and PARP expression were significantly increased in BEST1-pcDNA3.1 p.S142G and p.A146T group. Flow cytometry showed that the apoptosis rates were significantly increased in the BEST1-pcDNA3.1 p.V143F, p.S142G, and p.A146T group compared with the wild-type group.

Conclusions

For the first time, we found that the three mutations promoted RPE cell apoptosis. Furthermore, the results indicated that BEST1 mutations p.S142G and p.A146T may contribute apoptosis of RPE cells by targeting Caspase 3. Our observations suggested that the apoptosis of RPE cells may be one of the mechanisms in bestrophinopathies, which may provide a new potential therapeutic target for the treatment of this disease.
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Metadaten
Titel
Disease-causing mutations associated with bestrophinopathies promote apoptosis in retinal pigment epithelium cells
verfasst von
Tingting Gao
Chengqiang Tian
Hui Xu
Xin Tang
Lvzhen Huang
Mingwei Zhao
Publikationsdatum
07.06.2020
Verlag
Springer Berlin Heidelberg
Erschienen in
Graefe's Archive for Clinical and Experimental Ophthalmology / Ausgabe 10/2020
Print ISSN: 0721-832X
Elektronische ISSN: 1435-702X
DOI
https://doi.org/10.1007/s00417-020-04636-5

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