Institute of Genetic Medicine, Newcastle University, International Centre for Life, Central Parkway, Newcastle upon Tyne, NE1 3BZ, United Kingdom.
Renal Services, The Newcastle Hospitals NHS Foundation Trust, Newcastle upon Tyne, NE7 7DN, United Kingdom.
Sci Rep. 2019 Jul 25;9(1):10828. doi: 10.1038/s41598-019-47243-z.
Joubert syndrome (JBTS) is an incurable multisystem ciliopathy syndrome. The most commonly mutated gene in JBTS patients with a cerebello-retinal-renal phenotype is CEP290 (alias JBTS5). The encoded CEP290 protein localises to the proximal end of the primary cilium, in the transition zone, where it controls ciliary protein composition and signalling. We examined primary cilium structure and composition in fibroblast cells derived from homozygous and compound heterozygous JBTS5 patients with nonsense mutations in CEP290 and show that elongation of cilia, impaired ciliogenesis and ciliary composition defects are typical features in JBTS5 cells. Targeted skipping of the mutated exon c.5668 G > T using antisense oligonucleotide (ASO) therapy leads to restoration of CEP290 protein expression and functions at the transition zone in homozygous and compound heterozygous JBTS5 cells, allowing a rescue of both cilia morphology and ciliary composition. This study, by demonstrating that targeted exon skipping is able to rescue ciliary protein composition defects, provides functional evidence for the efficacy of this approach in the treatment of JBTS.
杰伯综合征（JBTS）是一种无法治愈的多系统纤毛病。具有小脑视网膜肾表型的 JBTS 患者中最常突变的基因是 CEP290（别名 JBTS5）。编码的 CEP290 蛋白定位于初级纤毛的近端，在过渡区，控制纤毛蛋白组成和信号。我们研究了同源和复合杂合 JBTS5 患者的成纤维细胞中的初级纤毛结构和组成，这些患者的 CEP290 中有无义突变，表现为纤毛伸长、纤毛发生受损和纤毛组成缺陷是 JBTS5 细胞的典型特征。使用反义寡核苷酸（ASO）治疗靶向突变外显子 c.5668G>T 的剪接，可以恢复同源和复合杂合 JBTS5 细胞中 CEP290 蛋白在过渡区的表达和功能，从而挽救纤毛形态和纤毛组成。这项研究通过证明靶向外显子跳跃能够挽救纤毛蛋白组成缺陷，为该方法治疗 JBTS 的有效性提供了功能证据。
