Abstract

Introduction: Cystic fibrosis (CF) is a life-limiting genetic disorder affecting approximately 70,000 people worldwide. Current burden of treatment is high. While the latest pharmaceutical innovation has benefitted many, patients with certain genotypes remain excluded. Gene editing has the potential to correct the underlying cause of disease for all patients, representing a permanent cure.

Areas covered: Various DNA editing-based strategies for treatment are currently being developed. Different strategies are called for based upon location of mutations (intronic vs. exonic), delivery mechanism of editing machinery, and cell type being targeted. Furthermore, the unique physiology of the CF lung presents a variety of barriers to delivery of CRISPR-Cas9 machinery.

Expert opinion: The most significant obstacle to the use of CRISPR-Cas9 in vivo is the fact that the most clinically relevant and accessible CF tissue, the airway epithelium, is made up of non-dividing cells where precise editing via homology-directed repair (HDR) does not occur; rather, potentially deleterious imprecise editing via non-homologous end joining (NHEJ) dominates. Future research should focus on the development of either more precise NHEJ-based approaches, access to airway basal cells, editing approaches that do not involve introducing genomic double strand breaks, and strategies with ex vivo edited cells.

摘要

简介：囊性纤维化（CF）是一种限制生命的遗传疾病，影响全世界约70,000人。当前的治疗负担很高。尽管最新的药物创新使许多人受益，但某些基因型的患者仍被排除在外。基因编辑具有纠正所有患者潜在病因的潜力，代表了永久的治愈方法。

涵盖领域：目前正在开发各种基于DNA编辑的治疗策略。根据突变的位置（内含子与外显子），编辑机制的传递机制以及靶向的细胞类型，需要采取不同的策略。此外，CF肺部独特的生理特性对CRISPR-Cas9机器的运输提出了多种障碍。

专家意见：体内使用CRISPR-Cas9的最大障碍是以下事实：临床上最相关，最易接近的CF组织（气道上皮）由非分裂细胞组成，这些细胞通过同源性定向修复进行精确编辑（ HDR）不会发生；相反，通过非同源末端连接（NHEJ）进行的潜在有害，不精确的编辑占主导地位。未来的研究应侧重于基于NHEJ的更精确方法的开发，通气道基础细胞的访问，不涉及引入基因组双链断裂的编辑方法以及离体编辑细胞的策略。