Figure 1.

iPSC-derived HIOs can be used to measure the function of Wt-CFTR and pharmacologically rescued F508del-CFTR as Fsk-induced organoid swelling. (a) Schematic depicting the generation of 3D Human Intestinal Organoids (HIOs) from patient-derive iPSCs. (b) Characterization of iPSC-derived CF and non-CF intestinal organoids. Immunofluorescence studies of iPSC-differentiated HIOs highlighting expression of intestinal cell markers CDX2 (red, intestinal marker), and ZO-1 (red, epithelial cell/tight junction marker). (c) Representative images of Fsk-induced swelling of F508del-CFTR expressing CF organoids and Wt-CFTR expressing (non-CF) organoids. CF (CF01) organoids were rescued chronically (24 h) with DMSO control or VX-809 (3 µM) and acutely stimulated with Fsk (10 µM) or Fsk and VX-770 (1 µM). (d) Box and whisker plot shows the change in organoid size post-Fsk-induced swelling (ΔA) relative to average organoid size at baseline (A₀) (* p = 0.0191, ** p = 0.0065, n ≥ 3 biological replicates. Each biological replicate = independent organoid passage, technical replicate = average of >30 organoids). Box plot depicts the median value and bounds depict IQR ranges with the whiskers defining the minima and maxima values.