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 approximately 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.
- Merkert, S.
- Schubert, M.
- Olmer, R.
- Engels, L.
- Radetzki, S.
- Veltman, M.
- Scholte, B. J.
- Zollner, J.
- Pedemonte, N.
- Galietta, L. J. V.
- von Kries, J. P.
- Martin, U.
Keywords
- *cftr
- *cystic fibrosis
- *differentiation to intestinal epithelia
- *genome engineering by TALENs
- *halide-sensitive eYFP
- *high-throughput drug screening
- *human iPSCs
