Ivacaftor Reverses Airway Mucus Abnormalities in a Rat Model Harboring a Humanized G551D-CFTR
Ivacaftor Reverses Airway Mucus Abnormalities in a Rat Model Harboring a Humanized G551D-CFTR
复制标题
DOI：
10.1164/rccm.202002-0369oc
复制
发表时间：
2020-11-01
影响因子：
24.7
通讯作者：
Rowe, Steven M.
中科院分区：
文献类型：
--
作者：
Birket, Susan E.;Davis, Joy M.;Rowe, Steven M.
基本原理：动物模型对于理解囊性纤维化（CF）肺病的特征、发病和进展具有高度的信息性。特别是，CFTR-/-大鼠揭示了CF的气道粘液缺陷特征的见解，但不能复制人类相关的CFTR目的：我们假设表达CFTR的人源化版本并携带依伐卡托敏感性变体G551 D的大鼠可用于测试CFTR调节剂对病理生理发展和校正的影响。在这项研究中，我们描述了一个人源化的CFTR大鼠表达的G551 D变体通过锌指核酸酶编辑的人互补DNA superexon，跨越外显子2-27，与59插入位点到大鼠基因刚刚超出内含子1。这种有针对性的插入利用内源性大鼠启动子，导致适当的表达相比，野生型animals. Measures和主要结果：上皮细胞的生物电表型概括了预期的CFTR活性，这是恢复与ivacaftor的情况。大气道缺陷，包括耗尽气道表面液体和纤毛层，延迟粘液运输率，增加粘液粘度，正常化后，管理ivacaftor.Conclusions：该模型是有用的了解疾病的机制和程度的病理逆转CFTR调制器。
Rationale: Animal models have been highly informative for understanding the characteristics, onset, and progression of cystic fibrosis (CF) lung disease. In particular, the CFTR-/- rat has revealed insights into the airway mucus defect characteristic of CF but does not replicate a human-relevant CFTR (cystic fibrosis transmembrane conductance regulator) variant.Objectives: We hypothesized that a rat expressing a humanized version of CFTR and harboring the ivacaftor-sensitive variant G551D could be used to test the impact of CFTR modulators on pathophysiologic development and correction.Methods: In this study, we describe a humanized-CFTR rat expressing the G551D variant obtained by zinc finger nuclease editing of a human complementary DNA superexon, spanning exon 2-27, with a 59 insertion site into the rat gene just beyond intron 1. This targeted insertion takes advantage of the endogenous rat promoter, resulting in appropriate expression compared with wildtype animals.Measurements and Main Results: The bioelectric phenotype of the epithelia recapitulates the expected absence of CFTR activity, which was restored with ivacaftor. Large airway defects, including depleted airway surface liquid and periciliary layers, delayed mucus transport rates, and increased mucus viscosity, were normalized after the administration of ivacaftor.Conclusions: This model is useful to understand the mechanisms of disease and the extent of pathology reversal with CFTR modulators.