Department of Biochemistry and Molecular Genetics, School of Medicine, University of Alabama at Birmingham, Birmingham, Alabama, United States of America.
Department of Pediatrics, School of Medicine, University of Alabama at Birmingham, Birmingham, Alabama, United States of America.
PLoS One. 2023 Nov 29;18(11):e0295009. doi: 10.1371/journal.pone.0295009. eCollection 2023.
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.
囊性纤维化 (CF) 治疗领域存在一个未满足的主要需求，即缺乏针对无义 CFTR 突变的有效治疗方法，而无义 CFTR 突变约影响 10%的 CF 患者。通过基因组编辑纠正无义 CFTR 突变代表了一种很有前途的治疗方法。在这项研究中，我们测试了基于 CRISPR 的新型基因组编辑方法——Prime 编辑，是否可以作为一种潜在的治疗方法来纠正无义 CFTR 突变。我们从 CFTR W1282X 突变纯合的 CF 患者中生成了 iPSC。我们证明 Prime 编辑可以纠正 iPSC 中的一个突变等位基因，这有效地恢复了 iPSC 衍生的气道上皮细胞和类器官中的 CFTR 功能。我们进一步证明，当 Prime 编辑机制通过 helper-dependent adenovirus (HDAd) 有效递送至 iPSC 衍生的气道上皮细胞时，Prime 编辑可能会直接修复这些细胞中的突变。总之，我们的数据表明，Prime 编辑可能有潜力应用于纠正 CFTR 突变，如 W1282X。
