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Mutant Best1 Expression and Impaired Phagocytosis in an iPSC Model of Autosomal Recessive Bestrophinopathy

Autosomal recessive bestrophinopathy (ARB) is caused by mutations in the gene BEST1 which encodes bestrophin 1 (Best1), an anion channel expressed in retinal pigment epithelial (RPE) cells. It has been hypothesized that ARB represents the human null phenotype for BEST1 and that this occurs due to no...

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Autores principales: Marmorstein, Alan D., Johnson, Adiv A., Bachman, Lori A., Andrews-Pfannkoch, Cynthia, Knudsen, Travis, Gilles, Benjamin J., Hill, Matthew, Gandhi, Jarel K., Marmorstein, Lihua Y., Pulido, Jose S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5852082/
https://www.ncbi.nlm.nih.gov/pubmed/29540715
http://dx.doi.org/10.1038/s41598-018-21651-z
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author Marmorstein, Alan D.
Johnson, Adiv A.
Bachman, Lori A.
Andrews-Pfannkoch, Cynthia
Knudsen, Travis
Gilles, Benjamin J.
Hill, Matthew
Gandhi, Jarel K.
Marmorstein, Lihua Y.
Pulido, Jose S.
author_facet Marmorstein, Alan D.
Johnson, Adiv A.
Bachman, Lori A.
Andrews-Pfannkoch, Cynthia
Knudsen, Travis
Gilles, Benjamin J.
Hill, Matthew
Gandhi, Jarel K.
Marmorstein, Lihua Y.
Pulido, Jose S.
author_sort Marmorstein, Alan D.
collection PubMed
description Autosomal recessive bestrophinopathy (ARB) is caused by mutations in the gene BEST1 which encodes bestrophin 1 (Best1), an anion channel expressed in retinal pigment epithelial (RPE) cells. It has been hypothesized that ARB represents the human null phenotype for BEST1 and that this occurs due to nonsense mediated decay (NMD). To test this hypothesis, we generated induced pluripotent stem cells (iPSCs) from a patient with ARB and her parents. After differentiation to retinal pigment epithelial (iPSC-RPE) cells, both BEST1 mRNA and Best1 protein expression were compared to controls. BEST1 mRNA expression levels, determined by quantitative PCR, were similar in ARB iPSC-RPE, parental cells, and genetically unrelated controls. Western blotting revealed that CRALBP and RPE65 were expressed within the range delineated by unrelated controls in iPSC-RPE from the ARB donor and her parents. Best1 protein was detected in different clones of ARB iPSC-RPE, but at reduced levels compared to all controls. When tested for the ability to phagocytose photoreceptor outer segments, ARB iPSC-RPE exhibited impaired internalization. These data suggest that impaired phagocytosis is a trait common to the bestrophinopathies. Furthermore, ARB is not universally the result of NMD and ARB, in this patient, is not due to the absence of Best1.
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spelling pubmed-58520822018-03-21 Mutant Best1 Expression and Impaired Phagocytosis in an iPSC Model of Autosomal Recessive Bestrophinopathy Marmorstein, Alan D. Johnson, Adiv A. Bachman, Lori A. Andrews-Pfannkoch, Cynthia Knudsen, Travis Gilles, Benjamin J. Hill, Matthew Gandhi, Jarel K. Marmorstein, Lihua Y. Pulido, Jose S. Sci Rep Article Autosomal recessive bestrophinopathy (ARB) is caused by mutations in the gene BEST1 which encodes bestrophin 1 (Best1), an anion channel expressed in retinal pigment epithelial (RPE) cells. It has been hypothesized that ARB represents the human null phenotype for BEST1 and that this occurs due to nonsense mediated decay (NMD). To test this hypothesis, we generated induced pluripotent stem cells (iPSCs) from a patient with ARB and her parents. After differentiation to retinal pigment epithelial (iPSC-RPE) cells, both BEST1 mRNA and Best1 protein expression were compared to controls. BEST1 mRNA expression levels, determined by quantitative PCR, were similar in ARB iPSC-RPE, parental cells, and genetically unrelated controls. Western blotting revealed that CRALBP and RPE65 were expressed within the range delineated by unrelated controls in iPSC-RPE from the ARB donor and her parents. Best1 protein was detected in different clones of ARB iPSC-RPE, but at reduced levels compared to all controls. When tested for the ability to phagocytose photoreceptor outer segments, ARB iPSC-RPE exhibited impaired internalization. These data suggest that impaired phagocytosis is a trait common to the bestrophinopathies. Furthermore, ARB is not universally the result of NMD and ARB, in this patient, is not due to the absence of Best1. Nature Publishing Group UK 2018-03-14 /pmc/articles/PMC5852082/ /pubmed/29540715 http://dx.doi.org/10.1038/s41598-018-21651-z Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Marmorstein, Alan D.
Johnson, Adiv A.
Bachman, Lori A.
Andrews-Pfannkoch, Cynthia
Knudsen, Travis
Gilles, Benjamin J.
Hill, Matthew
Gandhi, Jarel K.
Marmorstein, Lihua Y.
Pulido, Jose S.
Mutant Best1 Expression and Impaired Phagocytosis in an iPSC Model of Autosomal Recessive Bestrophinopathy
title Mutant Best1 Expression and Impaired Phagocytosis in an iPSC Model of Autosomal Recessive Bestrophinopathy
title_full Mutant Best1 Expression and Impaired Phagocytosis in an iPSC Model of Autosomal Recessive Bestrophinopathy
title_fullStr Mutant Best1 Expression and Impaired Phagocytosis in an iPSC Model of Autosomal Recessive Bestrophinopathy
title_full_unstemmed Mutant Best1 Expression and Impaired Phagocytosis in an iPSC Model of Autosomal Recessive Bestrophinopathy
title_short Mutant Best1 Expression and Impaired Phagocytosis in an iPSC Model of Autosomal Recessive Bestrophinopathy
title_sort mutant best1 expression and impaired phagocytosis in an ipsc model of autosomal recessive bestrophinopathy
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5852082/
https://www.ncbi.nlm.nih.gov/pubmed/29540715
http://dx.doi.org/10.1038/s41598-018-21651-z
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