Mutations of RNF213 are responsible for sporadic cerebral cavernous malformation and lead to a mulberry-like cluster in zebrafish
Mutations of RNF213 are responsible for sporadic cerebral cavernous malformation and lead to a mulberry-like cluster in zebrafish
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RNF213突变导致斑马鱼散发性脑海绵状血管瘤并导致桑葚样簇
DOI：
10.1177/0271678x20914996
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发表时间：
2020-04-04
影响因子：
6.3
通讯作者：
Sheng, Wenli
中科院分区：
文献类型：
--
作者：
Lin, Jing;Liang, Jie;Sheng, Wenli
尽管家族性脑海绵状血管瘤主要归因于三个CCM基因（KRIT1、CCM2和PDCD10），但在具有独特病变的散发性脑海绵状血管瘤病例中未发现突变，表明散发性脑海绵状血管瘤存在其他基因。为了筛选候选基因，我们对 31 名散发性脑海绵状血管瘤患者和 32 名健康对照者进行了全外显子组测序，并鉴定了 5 名受影响个体携带 6 个 RNF213 杂合有害突变，但健康个体中没有 RNF213 突变。为了进一步证实RNF213与脑海绵状血管瘤相关，我们培育了rnf213a纯合敲除斑马鱼，发现斑马鱼中rnf213a的突变导致了桑葚状的紊乱流动血管通道簇，这让人想起人类脑海绵状血管瘤。此外，我们通过转录组测序和 RT-qPCR 分析发现，由于 rnf213a 突变，kbtbd7 和 anxa6 显着下调。基于 kbtbd7 和 anxa6 的 mRNA 部分挽救的桑葚样表型，我们认为 rnf213a 通过下调 kbtbd7 和 anxa6 来促进桑葚样簇。总之，我们首次证明RNF213是散发性脑海绵状血管瘤的新候选基因，并且rnf213a的突变导致斑马鱼出现桑葚样簇。
Although familial forms of cerebral cavernous malformation are mainly attributed to three CCM genes (KRIT1, CCM2 and PDCD10), no mutation is identified in sporadic cerebral cavernous malformation cases with a unique lesion, indicating additional genes for sporadic cerebral cavernous malformation. To screen the candidate genes, we conducted whole exome sequencing in 31 sporadic cerebral cavernous malformation patients and 32 healthy controls, and identified 5 affected individuals carrying 6 heterozygous deleterious mutations in RNF213 but no RNF213 mutation in healthy individuals. To further confirm RNF213 was associated with cerebral cavernous malformation, we generated rnf213a homozygous knockout zebrafish and found mutation of rnf213a in zebrafish led to a mulberry-like cluster of disordered-flow vascular channels which was reminiscent of human cerebral cavernous malformation. In addition, we revealed kbtbd7 and anxa6 were significantly downregulated due to rnf213a mutation through transcriptomic sequencing and RT-qPCR analysis. Based on the mulberry-like phenotype partly rescued by mRNA of kbtbd7 as well as anxa6, we suggested that rnf213a promoted mulberry-like cluster via downregulation of kbtbd7 and anxa6. Altogether, we firstly demonstrate RNF213is a novel candidate gene for sporadic cerebral cavernous malformation and the mutation of rnf213a is responsible for the mulberry-like cluster in zebrafish.