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High-Throughput Functional Analysis of CFTR and Other Apically Localized Proteins in iPSC-Derived Human Intestinal Organoids

Induced Pluripotent Stem Cells (iPSCs) can be differentiated into epithelial organoids that recapitulate the relevant context for CFTR and enable testing of therapies targeting Cystic Fibrosis (CF)-causing mutant proteins. However, to date, CF-iPSC-derived organoids have only been used to study phar...

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Autores principales: Xia, Sunny, Bozóky, Zoltán, Di Paola, Michelle, Laselva, Onofrio, Ahmadi, Saumel, Jiang, Jia Xin, Pitstick, Amy L., Jiang, Chong, Rotin, Daniela, Mayhew, Christopher N., Jones, Nicola L., Bear, Christine E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8699884/
https://www.ncbi.nlm.nih.gov/pubmed/34943927
http://dx.doi.org/10.3390/cells10123419
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author Xia, Sunny
Bozóky, Zoltán
Di Paola, Michelle
Laselva, Onofrio
Ahmadi, Saumel
Jiang, Jia Xin
Pitstick, Amy L.
Jiang, Chong
Rotin, Daniela
Mayhew, Christopher N.
Jones, Nicola L.
Bear, Christine E.
author_facet Xia, Sunny
Bozóky, Zoltán
Di Paola, Michelle
Laselva, Onofrio
Ahmadi, Saumel
Jiang, Jia Xin
Pitstick, Amy L.
Jiang, Chong
Rotin, Daniela
Mayhew, Christopher N.
Jones, Nicola L.
Bear, Christine E.
author_sort Xia, Sunny
collection PubMed
description Induced Pluripotent Stem Cells (iPSCs) can be differentiated into epithelial organoids that recapitulate the relevant context for CFTR and enable testing of therapies targeting Cystic Fibrosis (CF)-causing mutant proteins. However, to date, CF-iPSC-derived organoids have only been used to study pharmacological modulation of mutant CFTR channel activity and not the activity of other disease-relevant membrane protein constituents. In the current work, we describe a high-throughput, fluorescence-based assay of CFTR channel activity in iPSC-derived intestinal organoids and describe how this method can be adapted to study other apical membrane proteins. Specifically, we show how this assay can be employed to study CFTR and ENaC channels and an electrogenic acid transporter in the same iPSC-derived intestinal tissue. This phenotypic platform promises to expand CF therapy discovery to include strategies that target multiple determinants of epithelial fluid transport.
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spelling pubmed-86998842021-12-24 High-Throughput Functional Analysis of CFTR and Other Apically Localized Proteins in iPSC-Derived Human Intestinal Organoids Xia, Sunny Bozóky, Zoltán Di Paola, Michelle Laselva, Onofrio Ahmadi, Saumel Jiang, Jia Xin Pitstick, Amy L. Jiang, Chong Rotin, Daniela Mayhew, Christopher N. Jones, Nicola L. Bear, Christine E. Cells Article Induced Pluripotent Stem Cells (iPSCs) can be differentiated into epithelial organoids that recapitulate the relevant context for CFTR and enable testing of therapies targeting Cystic Fibrosis (CF)-causing mutant proteins. However, to date, CF-iPSC-derived organoids have only been used to study pharmacological modulation of mutant CFTR channel activity and not the activity of other disease-relevant membrane protein constituents. In the current work, we describe a high-throughput, fluorescence-based assay of CFTR channel activity in iPSC-derived intestinal organoids and describe how this method can be adapted to study other apical membrane proteins. Specifically, we show how this assay can be employed to study CFTR and ENaC channels and an electrogenic acid transporter in the same iPSC-derived intestinal tissue. This phenotypic platform promises to expand CF therapy discovery to include strategies that target multiple determinants of epithelial fluid transport. MDPI 2021-12-04 /pmc/articles/PMC8699884/ /pubmed/34943927 http://dx.doi.org/10.3390/cells10123419 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Xia, Sunny
Bozóky, Zoltán
Di Paola, Michelle
Laselva, Onofrio
Ahmadi, Saumel
Jiang, Jia Xin
Pitstick, Amy L.
Jiang, Chong
Rotin, Daniela
Mayhew, Christopher N.
Jones, Nicola L.
Bear, Christine E.
High-Throughput Functional Analysis of CFTR and Other Apically Localized Proteins in iPSC-Derived Human Intestinal Organoids
title High-Throughput Functional Analysis of CFTR and Other Apically Localized Proteins in iPSC-Derived Human Intestinal Organoids
title_full High-Throughput Functional Analysis of CFTR and Other Apically Localized Proteins in iPSC-Derived Human Intestinal Organoids
title_fullStr High-Throughput Functional Analysis of CFTR and Other Apically Localized Proteins in iPSC-Derived Human Intestinal Organoids
title_full_unstemmed High-Throughput Functional Analysis of CFTR and Other Apically Localized Proteins in iPSC-Derived Human Intestinal Organoids
title_short High-Throughput Functional Analysis of CFTR and Other Apically Localized Proteins in iPSC-Derived Human Intestinal Organoids
title_sort high-throughput functional analysis of cftr and other apically localized proteins in ipsc-derived human intestinal organoids
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8699884/
https://www.ncbi.nlm.nih.gov/pubmed/34943927
http://dx.doi.org/10.3390/cells10123419
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