CCM2L (Cerebral Cavernous Malformation 2 Like) Deletion Aggravates Cerebral Cavernous Malformation Through Map3k3-KLF Signaling Pathway
CCM2L (Cerebral Cavernous Malformation 2 Like) Deletion Aggravates Cerebral Cavernous Malformation Through Map3k3-KLF Signaling Pathway
复制标题
CCM2L（脑海绵状血管瘤 2 样）缺失通过 Map3k3-KLF 信号通路加重脑海绵状血管瘤
DOI：
10.1161/strokeaha.120.031523
复制
发表时间：
2021-04-01
期刊：
影响因子：
8.3
通讯作者：
Zheng, Xiangjian
中科院分区：
文献类型：
--
作者：
Choi, Jaesung P.;Yang, Xi;Zheng, Xiangjian
补充数字内容可在正文中找到。背景和目的：脑海绵状血管畸形是一种常见的脑血管疾病。CCMS是年轻人中风、脑出血和神经功能障碍的主要原因。CCM1、CCM2和CCM3的功能缺失突变已被确定为导致人类CCM的原因。CCM2样蛋白(CCM2-like，Ccm21)与CCM2类似，主要表达于内皮细胞。CCM2L(类CCM2)与CCM2竞争结合CCM1，并且在血管发育过程中与CCM2有拮抗作用。CCM2L在CCM发病机制中的作用尚不清楚。方法：从诱导型CCM小鼠模型中分离脑内皮细胞，进行基因表达分析。微计算机断层成像分析Ccm21基因敲除小鼠(Ccm21-−/−)与Ccm1或Ccm2基因缺陷小鼠的基因杂交的CCM损伤负荷，以确定Ccm21在CCM发病机制中的作用。与Map3k3fl/fl小鼠进行遗传杂交，以确定Map3k3在Ccm2l促进的CCM形成中的作用。结果：我们发现CCM2缺陷小鼠脑内皮细胞中Ccm21的表达增加。对CCM患者标本的RNA-seq数据分析显示，CCM2L的表达呈增加趋势，并与Kruppel-like factor2/4(KLF2/4)的表达呈正相关。显微计算机断层扫描显示，CCM2基因缺陷小鼠的Ccm21基因缺失增加了CCM的损伤体积，但对病变数量没有影响。与损伤负荷增加相关的是，双基因敲除小鼠(CCM2和Ccm2l缺陷小鼠)脑内皮细胞KLF2/4mRNA表达显著高于对照组(CCM2缺陷小鼠)。内皮细胞中MAP3k3基因的半合子缺失减轻了双基因敲除中CCM的损伤负担。这些结果提示CCM2L在CCM发病机制中调节Map3k3-KLF信号通路。结论：CCM2L的缺失通过增加MAP3K3-KLF信号通路而加重CCM2缺陷小鼠模型CCM损伤的形成。我们的数据提示Ccm21表达增加是CCM发病的一种代偿机制。
Supplemental Digital Content is available in the text. Background and Purpose: Cerebral cavernous malformation (CCM) is a common cerebrovascular disease. CCMs are major causes of stroke, cerebral hemorrhage, and neurological deficits in young individuals. Loss-of-function mutations in CCM1, CCM2, and CCM3 have been identified to cause CCM in humans. Ccm2-like (Ccm2l) is a paralog of Ccm2 and is predominantly expressed in endothelial cells (ECs). CCM2L (CCM2-like) competes with CCM2 for binding to CCM1 and has been shown to have an antagonistic function to that of CCM2 during vascular development. The role of CCM2L in CCM pathogenesis is unknown. Methods: We isolated brain ECs from the inducible-CCM mouse models for gene expression analysis. Micro-computed tomography imaging was used to analyze CCM lesion burden from the genetic cross of Ccm2l knockout mice (Ccm2l−/−) with Ccm1 or Ccm2-deficient mice to determine the role of Ccm2l in CCM pathogenesis. Genetic crosses with Map3k3fl/fl mice were used to determine the role of Map3k3 in Ccm2l-facilitated CCM formation. Results: We demonstrated increased Ccm2l expression in brain ECs of Ccm2-deficient mice. Analysis of RNA-seq data from CCM patient samples revealed a trend of increased CCM2L expression and its positive correlation with Kruppel-like factor 2/4 (KLF2/4) expression. Micro-computed tomography revealed that the deletion of Ccm2l in Ccm2-deficient mice increased CCM lesion volume compared with that of controls but had no effect on lesion numbers. Correlating to the increased lesion burden, Klf2/4 mRNA expressions in brain ECs were significantly increased in double knockouts (Ccm2- and Ccm2l-deficient mice) compared with that of controls (Ccm2 deficient). Hemizygous deletion of Map3k3 in ECs relieved CCM lesion burden in the double knockouts. These results suggest that CCM2L regulates the Map3k3-KLF signaling pathway in CCM pathogenesis. Conclusions: Loss of CCM2L aggravates CCM lesion formation in the Ccm2-deficient mouse model through increased Map3k3-KLF signaling. Our data suggest that increased Ccm2l expression is a compensatory mechanism in CCM pathogenesis.