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

Abstract

Summary: 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. : In this article, Martin and colleagues demonstrate the development and conduction of a high-throughput Cl−/I− exchange assay using CF disease-specific iPSC-derived intestinal cells. They screened a library of ∼42,500 chemical compounds and validated primary hits, underlining that the advantages of complex iPSC-derived culture systems for disease modeling can be utilized for drug screening in a true HT format. Keywords: cystic fibrosis, human iPSCs, genome engineering by TALENs, CFTR, high-throughput drug screening, halide-sensitive eYFP, differentiation to intestinal epithelia