NRAS(Q61R) mutation in human endothelial cells causes vascular malformations.
NRAS(Q61R) mutation in human endothelial cells causes vascular malformations.
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DOI：
10.1007/s10456-022-09836-7
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发表时间：
2022-08
期刊：
影响因子：
9.8
通讯作者：
中科院分区：
文献类型：
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作者：
NRAS 的体细胞突变驱动黑色素瘤和其他癌症的发病机制，但它们在血管异常，特别是人类内皮细胞异常中的作用尚不清楚。本研究的目的是确定人内皮细胞中体细胞激活的 NRASQ61R 突变是否会诱导异常血管生成，并开发体外和体内模型来识别致病途径和测试抑制剂。在这里，我们在体外和体内血管生成模型中使用表达人内皮细胞的突变型 NRASQ61R 和野生型 NRAS (NRASWT)。这些研究表明，人内皮细胞中 NRASQ61R 的表达导致异常纺锤形形态转变、增殖和迁移增加。与 NRASWT 细胞相比，NRASQ61R 内皮细胞的 ERK 磷酸化增加，表明 MAPK/ERK 通路过度激活。 NRASQ61R 突变内皮细胞在 3D 纤维蛋白凝胶模型和体内裸鼠异种移植物中产生异常扩大的血管通道。这些研究表明 NRASQ61R 可以驱动人内皮细胞中的异常血管生成。 MAP 激酶抑制剂 U0126 治疗可阻止 NRASQ61R 内皮细胞向纺锤形形态的转变，而 mTOR 抑制剂雷帕霉素则不能。
Somatic mutations in NRAS drive the pathogenesis of melanoma and other cancers but their role in vascular anomalies and specifically human endothelial cells is unclear. The goals of this study were to determine whether the somatic-activating NRASQ61R mutation in human endothelial cells induces abnormal angiogenesis and to develop in vitro and in vivo models to identify disease-causing pathways and test inhibitors. Here, we used mutant NRASQ61R and wild-type NRAS (NRASWT) expressing human endothelial cells in in vitro and in vivo angiogenesis models. These studies demonstrated that expression of NRASQ61R in human endothelial cells caused a shift to an abnormal spindle-shaped morphology, increased proliferation, and migration. NRASQ61R endothelial cells had increased phosphorylation of ERK compared to NRASWT cells indicating hyperactivation of MAPK/ERK pathways. NRASQ61R mutant endothelial cells generated abnormal enlarged vascular channels in a 3D fibrin gel model and in vivo, in xenografts in nude mice. These studies demonstrate that NRASQ61R can drive abnormal angiogenesis in human endothelial cells. Treatment with MAP kinase inhibitor U0126 prevented the change to a spindle-shaped morphology in NRASQ61R endothelial cells, whereas mTOR inhibitor rapamycin did not.