Functional Gene Correction for Cystic Fibrosis in Lung Epithelial Cells Generated from Patient iPSCs.
Functional Gene Correction for Cystic Fibrosis in Lung Epithelial Cells Generated from Patient iPSCs.
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DOI：
10.1016/j.celrep.2015.07.062
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发表时间：
2015-09-01
期刊：
影响因子：
8.8
通讯作者：
Verma IM
中科院分区：
文献类型：
--
作者：
Firth AL;Menon T;Parker GS;Qualls SJ;Lewis BM;Ke E;Dargitz CT;Wright R;Khanna A;Gage FH;Verma IM
肺部疾病是美国的一个主要原因，目前的治疗方法仅用于控制症状。最常见的慢性且危及生命的肺部遗传性疾病是囊性纤维化（CF），由囊性纤维化跨膜调节因子（CFTR）突变引起。我们从携带 CFTR 基因中 F508 纯合缺失的 CF 患者中产生了诱导多能干细胞 (iPSC)，这导致 CFTR 向细胞膜的加工有缺陷。使用 CRISPR 将校正序列靶向内源 CFTR 基因组位点，结合完全可切除的选择系统，可以精确校正该突变，从而显着提高了校正效率。校正后的 iPSC 随后分化为成熟的气道上皮细胞，其中证明了正常 CFTR 表达和功能的恢复。这种基于同基因 iPSC 的 CF 模型系统可以适应新治疗方法的开发。
Lung disease is a major cause of death in the USA, with current therapeutic approaches only serving to manage symptoms. The most common chronic and life-threatening genetic disease of the lung is Cystic fibrosis (CF) caused by mutations in the cystic fibrosis transmembrane regulator (CFTR). We have generated induced pluripotent stem cells (iPSC) from CF patients carrying a homozygous deletion of F508 in the CFTR gene, which results in defective processing of CFTR to the cell membrane. This mutation was precisely corrected using CRISPR to target corrective sequences to the endogenous CFTR genomic locus, in combination with a completely excisable selection system which significantly improved the efficiency of this correction. The corrected iPSC were subsequently differentiated to mature airway epithelial cells where recovery of normal CFTR expression and function was demonstrated. This isogenic iPSC-based model system for CF could be adapted for the development of new therapeutic approaches.