Disease-causing mutations associated with bestrophinopathies promote apoptosis in retinal pigment epithelium cells
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GENETICS
Disease-causing mutations associated with bestrophinopathies promote apoptosis in retinal pigment epithelium cells Tingting Gao 1,2 & Chengqiang Tian 3 & Hui Xu 2 & Xin Tang 2 & Lvzhen Huang 2 & Mingwei Zhao 2 Received: 21 September 2019 / Revised: 17 February 2020 / Accepted: 24 February 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
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. Keywords BEST1 mutation . Caspase 3 . Human retinal pigment epithelium cell . Apoptosis . Bestrophinopathy . Pathogenesis
Introduction Tingting Gao and Chengqiang Tian contributed equally to this work. * Lvzhen Huang [email protected] * Mingwei Zhao [email protected] 1
Department of Ophthalmology, Peking University Third Hospital, Beijing, China
2
Department of Ophthalmology & Clinical Centre of Optometry, Eye diseases and optometry Institute, Beijing Key Laboratory of Diagnosis and Therapy of Retinal and Choroid Diseases, College of Optometry, Peking University Health Science Center, Peking University People’s Hospital, Xizhimen South Street 11, Xi Cheng District, Beijing 100044, China
3
Department of Ophthalmology, The First Affiliated Hospital of Dalian Medical University, Dalian, China
Mutations in BEST1 gene (OMIM 153700) have been found in association with various retinal degenerative diseases which are referred to a
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