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. 2023 Jun 19;24(12):10358. doi: 10.3390/ijms241210358

Figure 1.

Figure 1

Electrophysiological response of 2D monolayer rectal organoids. (A) Representative transepithelial current measurements (TCM) in colonoids from non-CF (WT-CFTR) control. (B) Representative Isc tracing of chloride conductance mediated by CFTR in response to fsk and VX-770 for from a F508del/F508del subject whose rectal organoids were grown in Transwell® filters. Cells were pre-incubated with DMSO (pink tracing) or VX-809 (blue tracing) and/or VX-661-VX-445 (green tracing). (C) Representative Ussing chamber recordings of transepithelial current measurements in A559T-colonoids pre-incubated with CFTR modulators (3 μM, 24 h, 37 °C) or vehicle (DMSO). Arrows indicate the addition of compounds: 100 µM apical amiloride (1. Amil), apical and basal addition of 10 µM forskolin (2. Fsk), apical and basal addition of 0.3 µM VX-770 (3. VX-770) and apical and basal addition of 20 µM CFTR-inhibition PPQ-102 (4. PPQ). (D) Isc response of cAMP-induced chloride currents by forskolin and augmented by VX-770 and CFTR modulators in rectal epithelial organoids of CF subject carrying the A559T variant. Values were normalized by the surface area of the filters in all the cases. Values are shown as mean (±SEM) of a minimum of three independent experiments, representing the maximum activation of A559T-CFTR colonoids after stimulation by fsk and VX-770 (0.3 µM). Kruskal-Wallis test ** p ˂ 0.005. The red dotted line indicates the mean value of currents registered for non-CF subjects (WT) in response to fsk. The blue dotted line indicates the mean value of currents registered for F508del-colonoids upon treatment with VX-445-VX-661-VX-770 (ETI) and the grey dotted line indicates the mean value of currents registered for F508del-colonoids upon treatment with VX-809-VX-770 (LI) both in response to fsk, used here as a reference.