Open reading frame correction using splice-switching antisense oligonucleotides for the treatment of cystic fibrosis.
Open reading frame correction using splice-switching antisense oligonucleotides for the treatment of cystic fibrosis.
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应用剪接开关反义寡核苷酸进行开放阅读框校正治疗囊性纤维化。
DOI：
10.1073/pnas.2114886119
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发表时间：
2022-01-18
影响因子：
11.1
通讯作者：
Hastings ML
中科院分区：
文献类型：
--
作者：
Michaels WE;Pena-Rasgado C;Kotaria R;Bridges RJ;Hastings ML
移码和无义突变是疾病治疗发展的主要问题。通过诱导外显子跳过来消除信使RNA中的这些突变是一种相对未被探索的治疗方法，尽管它已经显示出对一些疾病的希望。在这里，我们表明，通过诱导跳过与囊性纤维化(CF)相关的常见STOP突变，跳过它所在的外显子，导致开放阅读框架的恢复，并以一种预期对目前没有有效治疗选择的CF患者具有治疗作用的方式恢复CFTR蛋白功能。这些结果对CF界来说是一个重要的进步，但也对其他疾病有影响，在这些疾病中，终止性突变是导致功能障碍的原因。导致过早终止密码子(PTCs)的CFTR基因突变在囊性纤维化(CF)中很常见。这种突变类型会导致一种严重的疾病形式，可能是由于无义介导的mRNA衰变导致CFTR信使RNA(MRNA)低表达，以及产生无功能的截短CFTR蛋白。目前针对CF的治疗方法主要针对残留蛋白功能，对这些类型突变的患者效果较差，部分原因是CFTR蛋白水平较低。剪接开关反义寡核苷酸(ASO)旨在诱导外显子跳跃以恢复mRNA开放阅读框架，已在临床前和临床上显示出治疗许多疾病的前景。我们假设ASO介导的CFTR外显子23的跳过将恢复与终止外显子突变相关的CFTR活性，包括CFTRp.W1282X，这是CF中第五大最常见的突变。在这里，我们证明了缺乏编码外显子23的氨基酸的CFTR具有部分功能，并且对目前临床上使用的校正和调节药物有反应。ASO诱导的外显子23跳过挽救了从纯合子CFTR-W1282X患者分离的原代人支气管上皮细胞中CFTR的表达和氯电流。这些结果支持ASOS用于治疗CFTR23外显子I类突变的CF患者，这些突变导致CFTRmRNA不稳定和CFTR蛋白截断。
Frameshift and nonsense mutations pose a major problem for disease therapeutic development. Eliminating these mutations from the messenger RNA by inducing exon skipping is a relatively unexplored treatment approach, though it has shown promise for some diseases. Here, we show that eliminating a common stop mutation associated with cystic fibrosis (CF), by inducing the skipping of the exon it is located in, results in a restoration of the open reading frame and recovers CFTR protein function in a manner expected to be therapeutic in CF patients who don’t currently have effective treatment options. These results are an important advancement for the CF community but also have implications for other diseases where terminating mutations are responsible for dysfunction. CFTR gene mutations that result in the introduction of premature termination codons (PTCs) are common in cystic fibrosis (CF). This mutation type causes a severe form of the disease, likely because of low CFTR messenger RNA (mRNA) expression as a result of nonsense-mediated mRNA decay, as well as the production of a nonfunctional, truncated CFTR protein. Current therapeutics for CF, which target residual protein function, are less effective in patients with these types of mutations due in part to low CFTR protein levels. Splice-switching antisense oligonucleotides (ASOs), designed to induce skipping of exons in order to restore the mRNA open reading frame, have shown therapeutic promise preclinically and clinically for a number of diseases. We hypothesized that ASO-mediated skipping of CFTR exon 23 would recover CFTR activity associated with terminating mutations in the exon, including CFTR p.W1282X, the fifth most common mutation in CF. Here, we show that CFTR lacking the amino acids encoding exon 23 is partially functional and responsive to corrector and modulator drugs currently in clinical use. ASO-induced exon 23 skipping rescued CFTR expression and chloride current in primary human bronchial epithelial cells isolated from a homozygote CFTR-W1282X patient. These results support the use of ASOs in treating CF patients with CFTR class I mutations in exon 23 that result in unstable CFTR mRNA and truncations of the CFTR protein.
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影响因子：
14.9
作者：
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通讯作者：
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影响因子：
5.6
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通讯作者：
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