Prime editing-mediated correction of the CFTR W1282X mutation in iPSCs and derived airway epithelial cells

• Published in: PloS one Vol. 18; no. 11; p. e0295009
• Main Authors: Li, Chao, Liu, Zhong, Anderson, Justin, Liu, Zhongyu, Tang, Liping, Li, Yao, Peng, Ning, Chen, Jianguo, Liu, Xueming, Fu, Lianwu, Townes, Tim M, Rowe, Steven M, Bedwell, David M, Guimbellot, Jennifer, Zhao, Rui
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 29-11-2023; Public Library of Science (PLoS)
• Subjects: Biology and Life Sciences; Codon, Nonsense; Cystic fibrosis; Cystic Fibrosis - drug therapy; Cystic Fibrosis - therapy; Cystic Fibrosis Transmembrane Conductance Regulator - genetics; Development and progression; Epithelial Cells; Gene mutations; Health aspects; Humans; Induced Pluripotent Stem Cells; Medicine and Health Sciences; Mutation; Research and Analysis Methods; United States
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0295009

A major unmet need in the cystic fibrosis (CF) therapeutic landscape is the lack of effective treatments for nonsense CFTR mutations, which affect approximately 10% of CF patients. Correction of nonsense CFTR mutations via genomic editing represents a promising therapeutic approach. In this study, we tested whether prime editing, a novel CRISPR-based genomic editing method, can be a potential therapeutic modality to correct nonsense CFTR mutations. We generated iPSCs from a CF patient homozygous for the CFTR W1282X mutation. We demonstrated that prime editing corrected one mutant allele in iPSCs, which effectively restored CFTR function in iPSC-derived airway epithelial cells and organoids. We further demonstrated that prime editing may directly repair mutations in iPSC-derived airway epithelial cells when the prime editing machinery is efficiently delivered by helper-dependent adenovirus (HDAd). Together, our data demonstrated that prime editing may potentially be applied to correct CFTR mutations such as W1282X.