High-Throughput Screening for Modulators of CFTR Activity Based on Genetically Engineered Cystic Fibrosis Disease-Specific iPSCs


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. Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.

Authors Merkert S, Schubert M, Olmer R, Engels L, Radetzki S, Veltman M, Scholte BJ, Zöllner J, Pedemonte N, Galietta LJV, von Kries JP, Martin U
Journal Stem cell reports
Publication Date 2019 Jun 11;12(6):1389-1403
PubMed 31080112
PubMed Central PMC6565754
DOI 10.1016/j.stemcr.2019.04.014

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