Mislocalisation of BEST1 in iPSC-derived retinal pigment epithelial cells from a family with autosomal dominant vitreoretinochoroidopathy (ADVIRC)

Carter, David A.; Smart, Matthew; Letton, William; Ramsden, Conor; Nommiste, Britta; Chen, Li Li; Fynes, Kate; Muthiah, Manickam Nick; Goh, Pollyanna; Lane, Amelia; Powner, Michael B.; Webster, Andrew R.; Cruz, Lyndon da; Moore, Anthony T.; Coffey, Peter; Carr, Amanda-Jayne F.

doi:10.1038/srep33792

Cited by 27 publications

(31 citation statements)

References 51 publications

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“…For the first time, this study provides data on the effect of ADVIRC-associated mutation BEST1-V86M on BEST1 localization, protein expression and anion transport function. While localization of mutated BEST1 to the basolateral plasma membrane was established in our study in two independent patient cell lines well in agreement with results from overexpression studies in a MDCKII cell model [18], ADVIRC-associated mutation (c.704T>C; p.(V235A)) was previously reported to be mislocalized at least in part to the apical surface of hiPSC-RPEs from an ADVIRC patient [35]. These contradictory results could suggest that proper BEST1 localization or a failure to correctly traffic to the cell membrane is rather mutation-dependent than an ADVIRC-specific characteristic and further underscores the necessity to model additional ADVIRC mutations on the basis of the hiPSC-RPE cell culture model.…”

Section: Discussionsupporting

confidence: 90%

“…To analyze splicing effects of ADVIRC mutation p.(V86M) under more native conditions, PCR amplification of full-length BEST1 from hiPSC-RPE cDNA from control and the two ADVIRC patients revealed exclusively the correct amplicon of 1.7 kb indicating normal splicing of exon 4 (Supplementary Figure S3B). Sanger sequencing of the amplicon confirmed its homology to the correct BEST1 transcript, thereby supporting previous findings [35]. Next, we quantified BEST1 protein expression in whole-cell lysates of hiPSC-RPEs from control, BD, ADVIRC, and ARB genotypes by Western blot analysis.…”

Section: Localization Rna and Protein Expression Of Mutant Best1 Insupporting

confidence: 84%

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Mutation-Dependent Pathomechanisms Determine the Phenotype in the Bestrophinopathies

Nachtigal

Milenkovic

Brandl

et al. 2020

IJMS

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Best vitelliform macular dystrophy (BD), autosomal dominant vitreoretinochoroidopathy (ADVIRC), and the autosomal recessive bestrophinopathy (ARB), together known as the bestrophinopathies, are caused by mutations in the bestrophin-1 (BEST1) gene affecting anion transport through the plasma membrane of the retinal pigment epithelium (RPE). To date, while no treatment exists a better understanding of BEST1-related pathogenesis may help to define therapeutic targets. Here, we systematically characterize functional consequences of mutant BEST1 in thirteen RPE patient cell lines differentiated from human induced pluripotent stem cells (hiPSCs). Both BD and ARB hiPSC-RPEs display a strong reduction of BEST1-mediated anion transport function compared to control, while ADVIRC mutations trigger an increased anion permeability suggesting a stabilized open state condition of channel gating. Furthermore, BD and ARB hiPSC-RPEs differ by the degree of mutant protein turnover and by the site of subcellular protein quality control with adverse effects on lysosomal pH only in the BD-related cell lines. The latter finding is consistent with an altered processing of catalytic enzymes in the lysosomes. The present study provides a deeper insight into distinct molecular mechanisms of the three bestrophinopathies facilitating functional categorization of the more than 300 known BEST1 mutations that result into the distinct retinal phenotypes.

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Section: Discussionsupporting

confidence: 90%

Section: Localization Rna and Protein Expression Of Mutant Best1 Insupporting

confidence: 84%

Mutation-Dependent Pathomechanisms Determine the Phenotype in the Bestrophinopathies

Nachtigal

Milenkovic

Brandl

et al. 2020

IJMS

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“…No. 08‐0027) as previously described by Carter et al . BJ iPSC cells were maintained in TeSR‐E8 (StemCell Technologies, Cambridge, U.K.) and grown on Matrigel (Corning, St David's Park, U.K.)‐coated six‐well plates.…”

Section: Methodsmentioning

confidence: 99%

Phenotypic and Functional Characterization of Müller Glia Isolated from Induced Pluripotent Stem Cell-Derived Retinal Organoids: Improvement of Retinal Ganglion Cell Function upon Transplantation

Eastlake

Wang

Jayaram

et al. 2019

Stem Cells Translational Medicine

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Glaucoma is one of the leading causes of blindness, and there is an ongoing need for new therapies. Recent studies indicate that cell transplantation using Müller glia may be beneficial, but there is a need for novel sources of cells to provide therapeutic benefit. In this study, we have isolated Müller glia from retinal organoids formed by human induced pluripotent stem cells (hiPSCs) in vitro and have shown their ability to partially restore visual function in rats depleted of retinal ganglion cells by NMDA. Based on the present results, we suggest that Müller glia derived from retinal organoids formed by hiPSC may provide an attractive source of cells for human retinal therapies, to prevent and treat vision loss caused by retinal degenerative conditions. Stem Cells Translational Medicine 2019;8:775&784

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“…Best1 Q238R , another mutant associated with BVMD, was similarly reported to be mislocalized in iPSC-RPE (Milenkovic et al, 2015). A recent study by Carter et al analyzed Best1 localization in iPSC-RPE derived from a patient with ADVIRC and the associated BEST1 mutation V235A (Carter et al, 2016). …”

Section: Pathogenesis Of the Bestrophinopathiesmentioning

confidence: 99%

“…While Best1 V235A was found to be properly localized in MDCK cells (Johnson et al, 2014), Best1 was found to be mislocalized in these ADVIRC iPSC-RPE (Carter et al, 2016). This is further evidence that future trafficking studies should strive to analyze Best1 localization in a human RPE model (e.g., iPSC-RPE, fhRPE, or RPE in situ).…”

Section: Pathogenesis Of the Bestrophinopathiesmentioning

confidence: 99%

Bestrophin 1 and retinal disease

Johnson

Guziewicz

Lee

et al. 2017

Progress in Retinal and Eye Research

198

177

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Mutations in the gene BEST1 are causally associated with as many as five clinically distinct retinal degenerative diseases, which are collectively referred to as the “bestrophinopathies”. These five associated diseases are: Best vitelliform macular dystrophy, autosomal recessive bestrophinopathy, adult-onset vitelliform macular dystrophy, autosomal dominant vitreoretinochoroidopathy, and retinitis pigmentosa. The most common of these is Best vitelliform macular dystrophy. Bestrophin 1 (Best1), the protein encoded by the gene BEST1, has been the subject of a great deal of research since it was first identified nearly two decades ago. Today we know that Best1 functions as both a pentameric anion channel and a regulator of intracellular Ca2+ signaling. Best1 is an integral membrane protein which, within the eye, is uniquely expressed in the retinal pigment epithelium where it predominantly localizes to the basolateral plasma membrane. Within the brain, Best1 expression has been documented in both glial cells and astrocytes where it functions in both tonic GABA release and glutamate transport. The crystal structure of Best1 has revealed critical information about how Best1 functions as an ion channel and how Ca2+ regulates that function. Studies using animal models have led to critical insights into the physiological roles of Best1 and advances in stem cell technology have allowed for the development of patient-derived, “disease in a dish” models. In this article we review our knowledge of Best1 and discuss prospects for near-term clinical trials to test therapies for the bestrophinopathies, a currently incurable and untreatable set of diseases.

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Mislocalisation of BEST1 in iPSC-derived retinal pigment epithelial cells from a family with autosomal dominant vitreoretinochoroidopathy (ADVIRC)

Cited by 27 publications

References 51 publications

Mutation-Dependent Pathomechanisms Determine the Phenotype in the Bestrophinopathies

Mutation-Dependent Pathomechanisms Determine the Phenotype in the Bestrophinopathies

Phenotypic and Functional Characterization of Müller Glia Isolated from Induced Pluripotent Stem Cell-Derived Retinal Organoids: Improvement of Retinal Ganglion Cell Function upon Transplantation

Bestrophin 1 and retinal disease

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