iPSC-Derived Intestinal Organoids from Cystic Fibrosis Patients Acquire CFTR Activity upon TALEN-Mediated Repair of the p.F508del Mutation
Aarne Fleischer, Sara Vallejo‐Díez, José María Martín‐Fernández, Almudena Sánchez‐Gilabert, Mónica Castresana, Ángel del Pozo, Amaia Esquisabel, Sílvia Ávila, José Luis Castrillo, Eusebio Gaínza, José Luís Pedraz, Miguel Viñas, Daniel Bachiller
Abstract
gene, but one of them, phenylalanine residue at amino acid position 508 (p.F508del), a recessive allele, is responsible for the vast majority of CF cases worldwide. Here, we present the results of the application of genome-editing techniques to the restoration of CFTR activity in p.F508del patient-derived induced pluripotent stem cells (iPSCs). Gene-edited iPSCs were subsequently used to produce intestinal organoids on which the physiological activity of the restored gene was tested in forskolin-induced swelling tests. The seamless restoration of the p.F508del mutation resulted in normal expression of the mature CFTR glycoprotein, full recovery of CFTR activity, and a normal response of the repaired organoids to treatment with two approved CF therapies: VX-770 and VX-809.