Biochemical and Functional Characterization of RNF213 (Mysterin) R4810K, a Susceptibility Mutation of Moyamoya Disease, in Angiogenesis In Vitro and In Vivo.
Biochemical and Functional Characterization of RNF213 (Mysterin) R4810K, a Susceptibility Mutation of Moyamoya Disease, in Angiogenesis In Vitro and In Vivo.
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DOI：
10.1161/jaha.115.002146
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发表时间：
2015-06-30
影响因子：
5.4
通讯作者：
Koizumi A
中科院分区：
文献类型：
--
作者：
Kobayashi H;Matsuda Y;Hitomi T;Okuda H;Shioi H;Matsuda T;Imai H;Sone M;Taura D;Harada KH;Habu T;Takagi Y;Miyamoto S;Koizumi A
RNF 213（mysterin：rs 112735431）的P.R4810K是东亚人烟雾病（moyamoya disease，MMD）的易感基因。然而，RNF 213 R4810 K在MMD病因学中的作用尚不清楚。为了阐明RNF 213在已知血管生成途径中的作用，在用几种血管生成和抗血管生成因子（包括干扰素（IFN））处理的内皮细胞（EC）中分析RNF 213表达。IFN-β通过启动子中的信号转导子和转录激活子x上调RNF 213，并介导IFN-β的抗血管生成活性。RNF 213野生型（WT）过表达不能降低无IFN-β的血管生成，但RNF 213 R4810 K过表达可以。为了关联ATP酶的生化功能和RNF 213寡聚体形成与抗血管生成活性的作用，我们研究了AAA+模块中突变的影响。稳定寡聚化的步行者B基序（WEQ）的突变抑制血管生成，但不能启动寡聚化的AAA+模块缺失没有抑制血管生成。有趣的是，R4810 K，类似于WEQ，降低ATP酶活性，表明其通过稳定寡聚体的抗血管生成活性。为了证实RNF 213在体内上调的抗血管生成作用，将血管EC或平滑肌细胞特异性Rnf 213 R4757 K（R4810 K直系同源物）或WT转基因（Tg）小鼠暴露于缺氧。在EC特异性Rnf 213 R4757 K Tg小鼠中，缺氧抑制了脑血管生成，而在其他小鼠中没有抑制。本研究提示炎症信号作为环境因子和R4810 K载体对脑缺氧易感性的重要性。ATP与第一个AAA+结合的特异性抑制剂可能是MMD的有希望的治疗候选物。
P.R4810K of RNF213 (mysterin: rs112735431), which is an AAA+ ATPase, is the susceptibility polymorphism for moyamoya disease (MMD) in East Asians. However, the role of RNF213 R4810K in the etiology of MMD is unknown. To clarify the role of RNF213 in known angiogenic pathways, RNF213 expression was analyzed in endothelial cells (ECs) treated with several angiogenic and antiangiogenic factors, including interferons (IFNs). RNF213 was upregulated by IFN-β through signal transducer and activator of transcription x in the promoter and mediated antiangiogenic activity of IFN-β. RNF213 wild-type (WT) overexpression could not lower angiogenesis without IFN-β, but RNF213 R4810K overexpression could. To correlate biochemical function as ATPase and the role of RNF213 oligomer formation with antiangiogenic activity, we investigated the effects of mutations in the AAA+ module. A mutation of the Walker B motif (WEQ), which stabilizes oligomerization, inhibited angiogenesis, but AAA+ module deletion, which cannot initiate oligomerization, did not. Intriguingly, R4810K, similar to WEQ, decreased ATPase activity, suggesting its antiangiogenic activity through stabilizing oligomers. To confirm the antiangiogenic effect of RNF213 upregulation in vivo, vascular EC- or smooth muscle cell-specific Rnf213 R4757K (R4810K ortholog) or WT transgenic (Tg) mice were exposed to hypoxia. Cerebral angiogenesis by hypoxia was suppressed in EC-specific Rnf213 R4757K Tg mice, whereas it was not suppressed in other mice. This study suggests the importance of inflammatory signals as environmental factors and R4810K carriers for susceptibility to cerebral hypoxia. A specific inhibitor of ATP binding to the first AAA+ could be a promising therapeutic candidate for MMD.