Journal of Cystic Fibrosis The CF Canada-Sick Kids Program in individual CF therapy: A resource for the advancement of personalized medicine in CF  Paul D.W. Eckford, Jacqueline McCormack, Lise Munsie, Gengming He, Sanja Stanojevic, Sergio L. Pereira, Karen Ho, Julie Avolio, Claire Bartlett, Jin Ye Yang, Amy P. Wong, Leigh Wellhauser, Ling Jun Huan, Jia Xin Jiang, Hong Ouyang, Kai Du, Michelle Klingel, Lianna Kyriakopoulou, Tanja Gonska, Theo J. Moraes, Lisa J. Strug, Janet Rossant, Felix Ratjen, Christine E. Bear  Journal of Cystic Fibrosis  DOI: 10.1016/j.jcf.2018.03.013 Copyright © 2018 The Authors Terms and Conditions

Fig. 1 Composition of the CFIT resource. This figure shows the sample composition of the 100 cell line project. Samples are collected from F508del homozygous individuals prior to ORKAMBI® and individuals who are homozygous for rare mutations (some included in the list) are prioritized. Comprehensive clinical phenotype data is available for each donor and for those taking ORKAMBI®, clinical phenotypic assessment occurs at discrete time intervals as shown in Table 3. DNA is extracted from one blood sample in the clinical lab (Genome Diagnostics) using methods to ensure long fragments suitable for single molecule real time (SMRT) sequencing with the Pacific Biosciences (PacBio) technology at 30× coverage. Another blood sample is sent to the Centre for Commercialization of Regenerative Medicine (CCRM, https://ccrm.ca) for the generation and characterization of iPSCs. Nasal epithelial cell brushings from each nostril (2 samples) are obtained by JA and RNA extracted by Genome Diagnostics for one sample and from the other sample – cells are expanded and differentiated to a monolayer for functional studies of drug responses. Cell lines are stored and can be accessed by academic laboratories through The Centre for Applied Genomics (SickKids). Journal of Cystic Fibrosis DOI: (10.1016/j.jcf.2018.03.013) Copyright © 2018 The Authors Terms and Conditions

Fig. 2 A: Differentiation in an air liquid interface results in a polarized layer of cells with apical CFTR. Blue stain, DAPI (cell nuclei); red stain, β4 tubulin (cilia); green stain, CFTR. Subject shown is non-CF. The presence of cilia and apical CFTR indicate a well differentiated respiratory epithelial phenotype. B: Ussing chamber analysis shows pharmacological modification of CFTR related activity. Representative tracings are shown from a CF subject's cells. Upper panel shows baseline (DMSO control) responses and responses after 48 h incubation with VX-809 3 μM (& acute VX-770 1 μM) in a poorly responding nasal cell sample. Lower panel shows a responsive nasal cell sample. After amiloride (Amil) and VX-770 addition, forskolin (fsk) activates a negative transepithelial current, which can be blocked by CFTR inhibitor (CFTRInh-172). Note the forskolin-activated/CFTR inhibitor172-sensitive current present in the lower but not the upper panel. This demonstrates undetectable CFTR function at baseline but functional restoration post VX-809/VX-770 in only some F508del homozygous cell samples. Note: Ussing experiment is performed in open circuit mode. Currents are determined using Ohm's law. We measure bioelectric responses by the nasal cultures in open circuit to retain native electrochemical driving forces. Values in short circuit mode are generally higher than in open circuit mode due to a non-physiological electrochemical driving forces. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.) Journal of Cystic Fibrosis DOI: (10.1016/j.jcf.2018.03.013) Copyright © 2018 The Authors Terms and Conditions