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Stem Cell Reports. 2019 Jun 11;12(6):1389-1403. doi: 10.1016/j.stemcr.2019.04.014. Epub 2019 May 9.

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

Author information

1
Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation and Vascular Surgery, Hannover Medical School, 30625 Hannover, Germany; REBIRTH-Cluster of Excellence, Hannover Medical School, 30625 Hannover, Germany; Biomedical Research in Endstage and Obstructive Lung Disease (BREATH), Member of the German Center for Lung Research (DZL), 30625 Hannover, Germany.
2
Leibniz-Forschnungsinstitut für Molekulare Pharmakologie (FMP), 13125 Berlin, Germany.
3
ErasmusMC, Sophia Children's Hospital, Pediatric Pulmonology, 3015 AA Rotterdam, The Netherlands; Cell Biology Department Rotterdam, 3015 AA Rotterdam, The Netherlands.
4
UOC Genetica Medica, IRCCS Istituto Giannina Gaslini, 16147 Genova, Italy.
5
Telethon Institute of Genetics and Medicine (TIGEM), 80078 Pozzuoli, Italy.
6
Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation and Vascular Surgery, Hannover Medical School, 30625 Hannover, Germany; REBIRTH-Cluster of Excellence, Hannover Medical School, 30625 Hannover, Germany; Biomedical Research in Endstage and Obstructive Lung Disease (BREATH), Member of the German Center for Lung Research (DZL), 30625 Hannover, Germany. Electronic address: martin.ulrich@mh-hannover.de.

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:

CFTR; cystic fibrosis; differentiation to intestinal epithelia; genome engineering by TALENs; halide-sensitive eYFP; high-throughput drug screening; human iPSCs

PMID:
31080112
DOI:
10.1016/j.stemcr.2019.04.014
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