The role of alternative splicing in CEP290-related disease pathogenesis
The role of alternative splicing in CEP290-related disease pathogenesis
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选择性剪接在 CEP290 相关疾病发病机制中的作用
DOI：
10.1101/2022.03.03.22271834
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Taylor R
中科院分区：
文献类型：
--
作者：
Taylor R
原发性纤毛病是一组由原发性纤毛缺陷引起的遗传性发育障碍。CEP 290（290 kDa的中心体蛋白）突变是隐性纤毛病最常见的原因（发病率高达1：15，000）。致病性变体跨越这个大（93.2kb）54外显子基因的全长，引起范围从孤立的遗传性视网膜营养不良（IRD; Leber先天性黑蒙，LCA）到影响视网膜、肾和脑的多效范围的严重综合征性多器官纤毛病的表型。大多数致病性CEP 290变异体被预测为无效（37%无义，42%移码），但没有明确的基因型-表型关联。CEP 290基因编码外显子中几乎有一半（26/53）是同相“跳跃”外显子。位于跳跃外显子的变异体可以通过跳跃外显子从CEP 290转录本中去除，无义相关的改变剪接（NAS）被认为是减弱无义或移码CEP 290变异体致病性的一种机制。在此，我们利用生物信息学技术研究了CEP 290突变外显子对NAS的易感性。然后，我们使用CRISPR-Cas9技术对诱导多能干细胞（iPSC）中的CEP 290移码突变进行建模，并分析其对剪接和纤毛发生的影响。我们鉴定了外显子36，LCA突变的热点，作为NAS的强有力候选者，我们在表现出序列特异性外显子跳跃的突变iPSC中证实了这一点。外显子36跳跃不影响纤毛发生，与较大的移码突变体相反，该突变体显著降低了iPSC中纤毛的大小和发病率。我们认为，序列特异性NAS为CEP 290相关疾病的遗传多效性提供了分子基础。
Primary ciliopathies are a group of inherited developmental disorders resulting from defects in the primary cilium. Mutations inCEP290(Centrosomal protein of 290kDa) are the most frequent cause of recessive ciliopathies (incidence up to 1:15,000). Pathogenic variants span the full length of this large (93.2kb) 54 exon gene, causing phenotypes ranging from isolated inherited retinal dystrophies (IRDs; Leber Congenital Amaurosis, LCA) to a pleiotropic range of severe syndromic multi-organ ciliopathies affecting retina, kidney and brain. Most pathogenicCEP290variants are predicted null (37% nonsense, 42% frameshift), but there is no clear genotype-phenotype association. Almost half (26/53) of the coding exons inCEP290are in-phase “skiptic” (or skippable) exons. Variants located in skiptic exons could be removed fromCEP290transcripts by skipping the exon, and nonsense-associated altered splicing (NAS) has been proposed as a mechanism that attenuates the pathogenicity of nonsense or frameshiftCEP290variants. Here, we have usedin silicobioinformatic techniques to study the propensity ofCEP290skiptic exons for NAS. We then used CRISPR-Cas9 technology to modelCEP290frameshift mutations in induced pluripotent stem cells (iPSCs) and analysed their effects on splicing and ciliogenesis. We identified exon 36, a hotspot for LCA mutations, as a strong candidate for NAS that we confirmed in mutant iPSCs that exhibited sequence-specific exon skipping. Exon 36 skipping did not affect ciliogenesis, in contrast to a larger frameshift mutant that significantly decreased cilia size and incidence in iPSCs. We suggest that sequence-specific NAS provides the molecular basis of genetic pleiotropy forCEP290-related disorders.
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DOI：
10.1093/bioinformatics/btu638
发表时间：
2015-01-15
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Anders S;Pyl PT;Huber W
通讯作者：
Huber W
影响因子：
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Swaroop A
影响因子：
14.9
作者：
NCBI Resource Coordinators
通讯作者：
NCBI Resource Coordinators
DOI：
10.2215/cjn.05660517
发表时间：
2017-12-07
影响因子：
9.8
作者：
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DOI：
10.1140/epje/s10189-021-00031-y
发表时间：
2021-03-08
期刊：
The European physical journal. E, Soft matter
影响因子：
--
作者：
Hansen JN;Rassmann S;Stüven B;Jurisch-Yaksi N;Wachten D
通讯作者：
Wachten D