当前位置: X-MOL 学术Pediatr. Nephrol. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
The aminoglycoside geneticin permits translational readthrough of the CTNS W138X nonsense mutation in fibroblasts from patients with nephropathic cystinosis.
Pediatric Nephrology ( IF 3 ) Pub Date : 2018-11-09 , DOI: 10.1007/s00467-018-4094-0
Emma J Brasell 1 , LeeLee Chu 2 , Reyhan El Kares 2 , Jung Hwa Seo 2 , Robin Loesch 3 , Diana M Iglesias 4 , Paul Goodyer 1, 2, 5
Affiliation  

BACKGROUND Cystinosis is an ultrarare disorder caused by mutations of the cystinosin (CTNS) gene, encoding a cystine-selective efflux channel in the lysosomes of all cells of the body. Oral therapy with cysteamine reduces intralysosomal cystine accumulation and slows organ deterioration but cannot reverse renal Fanconi syndrome nor prevent the eventual need for renal transplantation. A definitive therapeutic remains elusive. About 15% of cystinosis patients worldwide carry one or more nonsense mutations that halt translation of the CTNS protein. Aminoglycosides such as geneticin (G418) can bind to the mammalian ribosome, relax translational fidelity, and permit readthrough of premature termination codons to produce full-length protein. METHODS To ascertain whether aminoglycosides permit readthrough of the most common CTNS nonsense mutation, W138X, we studied the effect of G418 on patient fibroblasts. RESULTS G418 treatment induced translational readthrough of CTNSW138X constructs transfected into HEK293 cells and expression of full-length endogenous CTNS protein in homozygous W138X fibroblasts. CONCLUSIONS Reduction in intracellular cystine indicates that the CTNS protein produced is functional as a cystine transporter. Interestingly, similar effects were seen even in W138X compound heterozygotes. These studies establish proof-of-principle for the potential of aminoglycosides to treat cystinosis and possibly other monogenic diseases caused by nonsense mutations.

中文翻译:

氨基糖苷遗传霉素可以使肾病性胱氨酸病患者的成纤维细胞中的CTNS W138X无意义突变进行翻译通读。

背景技术膀胱疾病是由胱氨酸(CTNS)基因突变引起的一种罕见病,该基因编码人体内所有细胞的溶酶体中的胱氨酸选择性外排通道。半胱胺的口服治疗减少了溶酶体中胱氨酸的积累,减缓了器官的恶化,但不能逆转肾Fanconi综合征,也不能防止最终需要进行肾移植。最终的治疗方法仍然难以捉摸。全世界约有15%的胱氨酸病患者携带一种或多种无意义的突变,这些突变会中断CTNS蛋白的翻译。氨基糖苷类如遗传霉素(G418)可以与哺乳动物核糖体结合,放松翻译保真度,并允许过早终止密码子的通读以产生全长蛋白。方法为了确定氨基糖苷类是否允许通读最常见的CTNS无意义突变W138X,我们研究了G418对患者成纤维细胞的作用。结果G418处理可诱导转染到HEK293细胞中的CTNSW138X构建体的翻译通透性,以及纯合W138X成纤维细胞中全长内源性CTNS蛋白的表达。结论细胞内胱氨酸的减少表明产生的CTNS蛋白起胱氨酸转运蛋白的作用。有趣的是,甚至在W138X化合物杂合子中也观察到了类似的效果。这些研究为氨基糖苷治疗胱氨酸和其他由无意义突变引起的单基因疾病的潜力建立了原理证明。结果G418处理可诱导转染到HEK293细胞中的CTNSW138X构建体的翻译通透性,以及纯合W138X成纤维细胞中全长内源性CTNS蛋白的表达。结论细胞内胱氨酸的减少表明产生的CTNS蛋白起胱氨酸转运蛋白的作用。有趣的是,甚至在W138X化合物杂合子中也观察到了类似的效果。这些研究为氨基糖苷治疗胱氨酸和其他由无意义突变引起的单基因疾病的潜力建立了原理证明。结果G418处理可诱导转染到HEK293细胞中的CTNSW138X构建体的翻译通透性,以及纯合W138X成纤维细胞中全长内源性CTNS蛋白的表达。结论细胞内胱氨酸的减少表明产生的CTNS蛋白起胱氨酸转运蛋白的作用。有趣的是,甚至在W138X化合物杂合子中也观察到了类似的效果。这些研究为氨基糖苷治疗胱氨酸和其他由无意义突变引起的单基因疾病的潜力建立了原理证明。
更新日期:2018-11-09
down
wechat
bug