Top

Graefe's Archive for Clinical and Experimental Ophthalmology

Published in:

01-10-2020 | Genetics

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

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

Published in: Graefe's Archive for Clinical and Experimental Ophthalmology | Issue 10/2020

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.

Boon CJ, Klevering BJ, Leroy BP et al (2009) The spectrum of ocular phenotypes caused by mutations in the BEST1 gene. Prog Retin Eye Res 28(3):187–205CrossRef

Tingting G, Jinfeng Q, Xiao J et al (2018) Clinical and mutation analysis of patients with best Vitelliform macular dystrophy or autosomal recessive bestrophinopathy in Chinese population. Biomed Res Int. https://doi.org/10.1155/2018/4582816

Masuda T, Esumi N (2010) SOX9, through interaction with microphthalmia-associated transcription factor (MITF) and OTX2, regulates BEST1 expression in the retinal pigment epithelium. J Biol Chem 285(35):26933–26944CrossRef

Hartzell HC, Qu Z, Yu K et al (2008) Molecular physiology of bestrophins: multifunctional membrane proteins linked to best disease and other retinopathies. Physiol Rev 88(2):639–672CrossRef

Milenkovic VM, Langmann T, Schreiber R et al (2008) Molecular evolution and functional divergence of the bestrophin protein family. BMC Evol Biol 8:72CrossRef

Xiao Q, Hartzell HC, Yu K (2010) Bestrophins and retinopathies. Pflugers Arch 460(2):559–569CrossRef

Johnson AA, Guziewicz KE, Lee CJ et al (2017) Bestrophin 1 and retinal disease. Prog Retin Eye Res 58:45–69CrossRef

Guziewicz KE, Sinha D, Gómez NM et al (2017) Bestrophinopathy: an RPE-photoreceptor interface. Prog Retin Eye Res 58:70–88CrossRef

Strauss O (2005) The retinal pigment epithelium in visual function. Physiol Rev 85(3):845–881CrossRef

10.

Lu T, Tengyang S, Ke X et al (2017) Screening of BEST1 gene in a Chinese cohort with Best vitelliform macular dystrophy or autosomal recessive bestrophinopathy. Invest Ophthalmol Vis Sci 58:3366–3375CrossRef

11.

Mazzoni F, Mao Y, Finnemann SC (2019) Advanced analysis of photoreceptor outer segment phagocytosis by RPE cells in culture. Methods Mol Biol 1834:95–108CrossRef

12.

Yao L, Yu Zhang YX et al (2017) Patient-specific mutations impair BESTROPHIN1's essential role in mediating Ca2+-dependent Cl-currents in human RPE. eLife. 6:e29914CrossRef

13.

Andreeva TD, Petrova SD, MIadenova K et al (2018) Effects of Ca2+, Glu and GABA on hBest1 and composite hBest1/POPC surface films. Colloids Surf B: Biointerfaces 161:192–199CrossRef

14.

Samuel W, Jaworski C, Postnikova OA et al (2017) Appropriately differentiated ARPE-19 cells regain phenotype and gene expression profiles similar to those of native RPE cells. Mol Vis 23:60–89PubMedPubMedCentral

15.

Adijanto J, Philp NJ (2014) Cultured primary human fetal retinal pigment epithelium (hfRPE) as a model for evaluating RPE metabolism. Exp Eye Res 126:77–84CrossRef

16.

Aridor M, Hannan LA (2000) Traffic jam: a compendium of human diseases that affect intracellular transport processes. Traffic. 1(11):836–851CrossRef

17.

Kausalya PJ, Amasheh S, Günzel D et al (2006) Disease-associated mutations affect intracellular traffic and paracellular Mg2+ transport function of claudin-16. J Clin Invest 116(4):878–891CrossRef

18.

Marquardt A, Stöhr H, Passmore LA et al (1998) Mutations in a novel gene, VMD2, encoding a protein of unknown properties cause juvenile-onset vitelliform macular dystrophy (Best's disease). Hum Mol Genet 7(9):1517–1525CrossRef

19.

Petrukhin K, Koisti MJ, Bakall B et al (1998) Identification of the gene responsible for Best macular dystrophy. Nat Genet 19(3):241–247CrossRef

20.

Fischmeister R, Hartzell HC (2005) Volume sensitivity of the bestrophin family of chloride channels. J Physiol 562:477–491CrossRef

21.

Qu Z, Wei RW, Mann W et al (2003) Two bestrophins cloned from Xenopus laevis oocytes express Ca(2+)-activated Cl(−) currents. J Biol Chem 278(49):49563–49572CrossRef

22.

Sun H, Tsunenari T, Yau KW et al (2002) The vitelliform macular dystrophy protein defines a new family of chloride channels. Proc Natl Acad Sci U S A 99(6):4008–4013CrossRef

23.

Nicholson DW, Ali A, Thornberry NA et al (1995) Identification and inhibition of the ICE/CED-3 protease necessary for mammalian apoptosis. Nature. 376(6535):37–43CrossRef

24.

Fernandes-Alnemri T, Litwack G, Alnemri ES et al (1994) CPP32, a novel human apoptotic protein with homology to Caenorhabditis elegans cell death protein Ced-3 and mammalian interleukin-1 beta-converting enzyme. J Biol Chem 269(49):30761–30764PubMed

25.

Shaoju J, Yang Y, Ma L et al (2017) In vivo and in vitro induction of the apoptotic effects of oxysophoridine on colorectal cancer cells via the Bcl-2/Bax/caspase-3 signaling pathway. Oncol Lett 14(6):8000–8006

26.

Yu SW, Wang H, Poitras MF et al (2002) Mediation of poly(ADP-ribose) polymerase-1-dependent cell death by apoptosis-inducing factor. Science. 297(5579):259–263CrossRef

27.

Kumar JS, Suman S, Chandna S (2017) Radioresistant Sf9 insect cells readily undergo an intrinsic mode of apoptosis in response to histone deacetylase (HDAC) inhibition. Mol Cell Biochem. https://doi.org/10.1007/s11010-017-3245-0

28.

Alnemri ES, Robertson NM, Fernandes TF et al (1992) Overexpressed full-length human BCL2 extends the survival of baculovirus-infected Sf9 insect cells. Proc Natl Acad Sci U S A 89(16):7295–7299CrossRef

29.

Marmorstein AD, Johnson AA, Bachman LA et al (2018) Mutant BEST1 expression and impaired phagocytosis in an iPSC model of autosomal recessive bestrophinopathy. Sci Rep 8(1):4487CrossRef

Title: Disease-causing mutations associated with bestrophinopathies promote apoptosis in retinal pigment epithelium cells
Authors: Tingting Gao
Chengqiang Tian
Hui Xu
Xin Tang
Lvzhen Huang
Mingwei Zhao
Publication date: 01-10-2020
Publisher: Springer Berlin Heidelberg
Published in: Graefe's Archive for Clinical and Experimental Ophthalmology / Issue 10/2020
Print ISSN: 0721-832X
Electronic ISSN: 1435-702X
DOI: https://doi.org/10.1007/s00417-020-04636-5

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

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

Abstract

Purpose

Methods

Results

Conclusions

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 10/2020

Airborne pathogen projection during ophthalmic examination

Tracking postoperative head positioning in endothelial keratoplasty using a head positioning sensor

Efficacy of intracameral preservative-free triamcinolone acetonide in pediatric cataract surgery: a meta-analysis

Balancing orbital volume reduction and redistribution for a tailored surgical treatment in Graves’ ophthalmopathy

Altered microvascular density in patients with systemic lupus erythematosus treated with hydroxychloroquine—an optical coherence tomography angiography study

Evaluation of retinal vessel quantity within individual retinal structural layers using optical coherence tomography angiography