Antisense Oligonucleotide-Based Splicing Correction in Individuals with Leber Congenital Amaurosis due to Compound Heterozygosity for the c.2991+1655A>G Mutation in CEP290.
Antisense Oligonucleotide-Based Splicing Correction in Individuals with Leber Congenital Amaurosis due to Compound Heterozygosity for the c.2991+1655A>G Mutation in CEP290.
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DOI：
10.3390/ijms19030753
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发表时间：
2018-03-07
影响因子：
5.6
通讯作者：
Garanto A
中科院分区：
文献类型：
--
作者：
Duijkers L;van den Born LI;Neidhardt J;Bax NM;Pierrache LHM;Klevering BJ;Collin RWJ;Garanto A
莱伯先天性黑朦（LCA）是一种罕见的遗传性视网膜疾病，影响全球约1:50,000人。到目前为止，有25个基因突变与LCA相关，其中CEP290（编码中心体蛋白290 kDa）是最常见的突变基因。最常见的导致lca的CEP290突变，c.2991+1655A>G，导致假外显子插入到不同比例的CEP290转录本中。我们之前证明了反义寡核苷酸（AONs）对纯合携带这种突变的患者具有很高的治疗潜力，尽管迄今为止，尚不清楚挽救一个等位基因是否足以恢复CEP290的功能。在这里，我们评估了这种突变的复合杂合样本在RNA、蛋白质和细胞水平上的AON疗效，以及CEP290的蛋白质截断突变。我们证明AONs可以有效地恢复剪接并增加蛋白质水平。然而，由于患者来源的细胞系之间调解的高度可变性，AONs的功效在细胞水平上更难评估。这一观察结果表明，第二个等位基因的严重性以及每个个体中可能存在的其他遗传变异的重要性。总的来说，AONs似乎是治疗cep290相关LCA的一种很有前途的工具，不仅适用于c.2991+1655A>G变异的纯合携带者，也适用于复合杂合携带者。
Leber congenital amaurosis (LCA) is a rare inherited retinal disorder affecting approximately 1:50,000 people worldwide. So far, mutations in 25 genes have been associated with LCA, with CEP290 (encoding the Centrosomal protein of 290 kDa) being the most frequently mutated gene. The most recurrent LCA-causing CEP290 mutation, c.2991+1655A>G, causes the insertion of a pseudoexon into a variable proportion of CEP290 transcripts. We previously demonstrated that antisense oligonucleotides (AONs) have a high therapeutic potential for patients homozygously harbouring this mutation, although to date, it is unclear whether rescuing one single allele is enough to restore CEP290 function. Here, we assessed the AON efficacy at RNA, protein and cellular levels in samples that are compound heterozygous for this mutation, together with a protein-truncating mutation in CEP290. We demonstrate that AONs can efficiently restore splicing and increase protein levels. However, due to a high variability in ciliation among the patient-derived cell lines, the efficacy of the AONs was more difficult to assess at the cellular level. This observation points towards the importance of the severity of the second allele and possibly other genetic variants present in each individual. Overall, AONs seem to be a promising tool to treat CEP290-associated LCA, not only in homozygous but also in compound heterozygous carriers of the c.2991+1655A>G variant.
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影响因子：
--
作者：
Plotnikova, Olga V.;Pugacheva, Elena N.;Golemis, Erica A.
通讯作者：
Golemis, Erica A.
DOI：
10.1038/mtna.2012.21
发表时间：
2012-06-26
期刊：
Molecular therapy. Nucleic acids
影响因子：
--
作者：
通讯作者：
--
影响因子：
8.8
作者：
Shimada H;Lu Q;Insinna-Kettenhofen C;Nagashima K;English MA;Semler EM;Mahgerefteh J;Cideciyan AV;Li T;Brooks BP;Gunay-Aygun M;Jacobson SG;Cogliati T;Westlake CJ;Swaroop A
通讯作者：
Swaroop A
影响因子：
8.8
作者：
Bonifert, Tobias;Menendez, Irene Gonzalez;Wissinger, Bernd
通讯作者：
Wissinger, Bernd
影响因子：
4.4
作者：
Murray, Susan F.;Jazayeri, Ali;McCaleb, Michael L.
通讯作者：
McCaleb, Michael L.