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Patient-specific iPSC-derived photoreceptor precursor cells as a means to investigate retinitis pigmentosa

Next-generation and Sanger sequencing were combined to identify disease-causing USH2A mutations in an adult patient with autosomal recessive RP. Induced pluripotent stem cells (iPSCs), generated from the patient’s keratinocytes, were differentiated into multi-layer eyecup-like structures with featur...

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Autores principales: Tucker, Budd A, Mullins, Robert F, Streb, Luan M, Anfinson, Kristin, Eyestone, Mari E, Kaalberg, Emily, Riker, Megan J, Drack, Arlene V, Braun, Terry A, Stone, Edwin M
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3755341/
https://www.ncbi.nlm.nih.gov/pubmed/23991284
http://dx.doi.org/10.7554/eLife.00824
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author Tucker, Budd A
Mullins, Robert F
Streb, Luan M
Anfinson, Kristin
Eyestone, Mari E
Kaalberg, Emily
Riker, Megan J
Drack, Arlene V
Braun, Terry A
Stone, Edwin M
author_facet Tucker, Budd A
Mullins, Robert F
Streb, Luan M
Anfinson, Kristin
Eyestone, Mari E
Kaalberg, Emily
Riker, Megan J
Drack, Arlene V
Braun, Terry A
Stone, Edwin M
author_sort Tucker, Budd A
collection PubMed
description Next-generation and Sanger sequencing were combined to identify disease-causing USH2A mutations in an adult patient with autosomal recessive RP. Induced pluripotent stem cells (iPSCs), generated from the patient’s keratinocytes, were differentiated into multi-layer eyecup-like structures with features of human retinal precursor cells. The inner layer of the eyecups contained photoreceptor precursor cells that expressed photoreceptor markers and exhibited axonemes and basal bodies characteristic of outer segments. Analysis of the USH2A transcripts of these cells revealed that one of the patient’s mutations causes exonification of intron 40, a translation frameshift and a premature stop codon. Western blotting revealed upregulation of GRP78 and GRP94, suggesting that the patient’s other USH2A variant (Arg4192His) causes disease through protein misfolding and ER stress. Transplantation into 4-day-old immunodeficient Crb1(−/−) mice resulted in the formation of morphologically and immunohistochemically recognizable photoreceptor cells, suggesting that the mutations in this patient act via post-developmental photoreceptor degeneration. DOI: http://dx.doi.org/10.7554/eLife.00824.001
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spelling pubmed-37553412013-08-29 Patient-specific iPSC-derived photoreceptor precursor cells as a means to investigate retinitis pigmentosa Tucker, Budd A Mullins, Robert F Streb, Luan M Anfinson, Kristin Eyestone, Mari E Kaalberg, Emily Riker, Megan J Drack, Arlene V Braun, Terry A Stone, Edwin M eLife Developmental Biology and Stem Cells Next-generation and Sanger sequencing were combined to identify disease-causing USH2A mutations in an adult patient with autosomal recessive RP. Induced pluripotent stem cells (iPSCs), generated from the patient’s keratinocytes, were differentiated into multi-layer eyecup-like structures with features of human retinal precursor cells. The inner layer of the eyecups contained photoreceptor precursor cells that expressed photoreceptor markers and exhibited axonemes and basal bodies characteristic of outer segments. Analysis of the USH2A transcripts of these cells revealed that one of the patient’s mutations causes exonification of intron 40, a translation frameshift and a premature stop codon. Western blotting revealed upregulation of GRP78 and GRP94, suggesting that the patient’s other USH2A variant (Arg4192His) causes disease through protein misfolding and ER stress. Transplantation into 4-day-old immunodeficient Crb1(−/−) mice resulted in the formation of morphologically and immunohistochemically recognizable photoreceptor cells, suggesting that the mutations in this patient act via post-developmental photoreceptor degeneration. DOI: http://dx.doi.org/10.7554/eLife.00824.001 eLife Sciences Publications, Ltd 2013-08-27 /pmc/articles/PMC3755341/ /pubmed/23991284 http://dx.doi.org/10.7554/eLife.00824 Text en Copyright © 2013, Tucker et al http://creativecommons.org/licenses/by/3.0/ This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Developmental Biology and Stem Cells
Tucker, Budd A
Mullins, Robert F
Streb, Luan M
Anfinson, Kristin
Eyestone, Mari E
Kaalberg, Emily
Riker, Megan J
Drack, Arlene V
Braun, Terry A
Stone, Edwin M
Patient-specific iPSC-derived photoreceptor precursor cells as a means to investigate retinitis pigmentosa
title Patient-specific iPSC-derived photoreceptor precursor cells as a means to investigate retinitis pigmentosa
title_full Patient-specific iPSC-derived photoreceptor precursor cells as a means to investigate retinitis pigmentosa
title_fullStr Patient-specific iPSC-derived photoreceptor precursor cells as a means to investigate retinitis pigmentosa
title_full_unstemmed Patient-specific iPSC-derived photoreceptor precursor cells as a means to investigate retinitis pigmentosa
title_short Patient-specific iPSC-derived photoreceptor precursor cells as a means to investigate retinitis pigmentosa
title_sort patient-specific ipsc-derived photoreceptor precursor cells as a means to investigate retinitis pigmentosa
topic Developmental Biology and Stem Cells
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3755341/
https://www.ncbi.nlm.nih.gov/pubmed/23991284
http://dx.doi.org/10.7554/eLife.00824
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