High-throughput screening for modulators of CFTR activity based on genetically engineered cystic fibrosis disease-specific iPSCs

Merkert, S.; Schubert, M.; Olmer, R.; Engels, L.; Radetzki, S.; Veltman, M.; Scholte, B.J.; Zöllner, J.; Pedemonte, N.; Galietta, L.J.V.; von Kries, J.P.; Martin, U.

High-throughput screening for modulators of CFTR activity based on genetically engineered cystic fibrosis disease-specific iPSCs

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Item Type:	Article
Title:	High-throughput screening for modulators of CFTR activity based on genetically engineered cystic fibrosis disease-specific iPSCs
Creators Name:	Merkert, S., Schubert, M., Olmer, R., Engels, L., Radetzki, S., Veltman, M., Scholte, B.J., Zöllner, J., Pedemonte, N., Galietta, L.J.V., von Kries, J.P. and Martin, U.
Abstract:	Organotypic culture systems from disease-specific induced pluripotent stem cells (iPSCs) exhibit obvious advantages compared with immortalized cell lines and primary cell cultures, but implementation of iPSC-based high-throughput (HT) assays is still technically challenging. Here, we demonstrate the development and conduction of an organotypic HT Cl(-)/I(-) exchange assay using cystic fibrosis (CF) disease-specific iPSCs. The introduction of a halide-sensitive YFP variant enabled automated quantitative measurement of Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) function in iPSC-derived intestinal epithelia. CFTR function was partially rescued by treatment with VX-770 and VX-809, and seamless gene correction of the p.Phe508del mutation resulted in full restoration of CFTR function. The identification of a series of validated primary hits that improve the function of p.Phe508del CFTR from a library of ∼42,500 chemical compounds demonstrates that the advantages of complex iPSC-derived culture systems for disease modeling can also be utilized for drug screening in a true HT format.
Keywords:	Cystic Fibrosishuman iPSCs, Genome Engineering by TALENs, CFTR, High-Throughput Drug Screening, Halide-Sensitive eYFP, Differentiation to Intestinal Epithelia
Source:	Stem Cell Reports
ISSN:	2213-6711
Publisher:	Cell Press / Elsevier
Volume:	12
Number:	6
Page Range:	1389-1403
Date:	11 June 2019
Official Publication:	https://doi.org/10.1016/j.stemcr.2019.04.014
PubMed:	View item in PubMed

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