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NPJ Genom Med. 2017 Apr 14;2:12. doi: 10.1038/s41525-017-0015-6.

Phenotypic profiling of CFTR modulators in patient-derived respiratory epithelia.

Author information

1
Department of Physiology, University of Toronto, Toronto, ON, Canada.
2
Programme in Molecular Medicine, Research Institute, Hospital for Sick Children, Toronto, ON, Canada.
3
Department of Biochemistry, University of Toronto, Toronto, ON, Canada.
4
Programme in Developmental and Stem Cell Biology, Research Institute, Hospital for Sick Children, Toronto, ON, Canada.
5
Programme in Translational Medicine, Research Institute, Hospital for Sick Children, Toronto, ON, Canada.
6
Department of Paediatrics, University of Toronto, Toronto, ON, Canada.
7
Firestone Institute for Respiratory Health, Division of Respirology, McMaster University, Hamilton, ON, Canada.
8
Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada.
9
Programme in Genetics and Genome Biology, Research Institute, The Hospital for Sick Children, Toronto, ON, Canada.

Abstract

Pulmonary disease is the major cause of morbidity and mortality in patients with cystic fibrosis, a disease caused by mutations in the Cystic Fibrosis Transmembrane conductance Regulator (CFTR) gene. Heterogeneity in CFTR genotype-phenotype relationships in affected individuals plus the escalation of drug discovery targeting specific mutations highlights the need to develop robust in vitro platforms with which to stratify therapeutic options using relevant tissue. Toward this goal, we adapted a fluorescence plate reader assay of apical CFTR-mediated chloride conductance to enable profiling of a panel of modulators on primary nasal epithelial cultures derived from patients bearing different CFTR mutations. This platform faithfully recapitulated patient-specific responses previously observed in the "gold-standard" but relatively low-throughput Ussing chamber. Moreover, using this approach, we identified a novel strategy with which to augment the response to an approved drug in specific patients. In proof of concept studies, we also validated the use of this platform in measuring drug responses in lung cultures differentiated from cystic fibrosis iPS cells. Taken together, we show that this medium throughput assay of CFTR activity has the potential to stratify cystic fibrosis patient-specific responses to approved drugs and investigational compounds in vitro in primary and iPS cell-derived airway cultures.

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