Nidogen-2 Maintains the Contractile Phenotype of Vascular Smooth Muscle Cells and Prevents Neointima Formation via Bridging Jagged1-Notch3 Signaling
Nidogen-2 Maintains the Contractile Phenotype of Vascular Smooth Muscle Cells and Prevents Neointima Formation via Bridging Jagged1-Notch3 Signaling
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Nidogen-2 通过桥接 Jagged1-Notch3 信号维持血管平滑肌细胞的收缩表型并防止新内膜形成
DOI：
10.1161/circulationaha.120.053361
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发表时间：
2021-10-12
期刊：
影响因子：
37.8
通讯作者：
Kong, Wei
中科院分区：
文献类型：
--
作者：
Mao, Chenfeng;Ma, Zihan;Kong, Wei
补充数字内容可在文本中找到。背景资料：细胞外基质（ECM）微环境如何调节血管平滑肌细胞（VSMCs）的收缩表型并赋予血管稳态仍然是一个谜。研究方法：为了探索维持VSMC收缩表型的关键ECM蛋白，我们应用蛋白质-蛋白质相互作用网络分析来探索与VSMC表型相关的新型ECM蛋白。通过结合体外和体内遗传小鼠血管损伤模型，我们确定巢蛋白-2，基底膜糖蛋白，作为一个关键的ECM蛋白，维持血管平滑肌细胞的身份。结果：我们收集了一个VSMC表型相关的基因数据集，通过使用基因本体注释结合文献检索。ECM蛋白基因和来自VSMC表型相关基因数据集的基因之间的蛋白质-蛋白质相互作用的计算分析揭示了候选基因巢蛋白-2，一种参与VSMC表型调节的基底膜糖蛋白。事实上，nidogen-2缺陷的VSMC在体外表现出收缩表型的丧失，与野生型小鼠相比，nidogen-2-/-小鼠表现出加重的钢丝损伤后颈动脉新生内膜形成。进一步的生物信息学分析、免疫共沉淀试验和荧光素酶试验显示，巢蛋白-2与常规Notch配体Jagged 1特异性相互作用。Nidogen-2通过Jagged 1-Notch 3信号而不是Notch 1或Notch 2信号维持VSMC收缩表型。巢蛋白-2增强Jagged 1和Notch 3的相互作用和随后的Notch 3激活。相反，锯齿状蛋白1和刻缺蛋白3的相互作用，信号激活，锯齿状蛋白1触发的VSMC分化显着抑制巢蛋白-2缺陷的VSMC。因此，与野生型小鼠相比，巢蛋白-2-/-小鼠中Jagged 1过表达对新生内膜形成的抑制作用减弱。结论：Nidogen-2通过Jagged 1-Notch 3信号通路维持血管平滑肌细胞的收缩表型。Nidogen-2是Jagged 1-Notch 3信号传导所必需的。
Supplemental Digital Content is available in the text. Background: How the extracellular matrix (ECM) microenvironment modulates the contractile phenotype of vascular smooth muscle cells (VSMCs) and confers vascular homeostasis remains elusive. Methods: To explore the key ECM proteins in the maintenance of the contractile phenotype of VSMCs, we applied protein-protein interaction network analysis to explore novel ECM proteins associated with the VSMC phenotype. By combining in vitro and in vivo genetic mice vascular injury models, we identified nidogen-2, a basement membrane glycoprotein, as a key ECM protein for maintenance of vascular smooth muscle cell identity. Results: We collected a VSMC phenotype–related gene dataset by using Gene Ontology annotation combined with a literature search. A computational analysis of protein-protein interactions between ECM protein genes and the genes from the VSMC phenotype–related gene dataset revealed the candidate gene nidogen-2, a basement membrane glycoprotein involved in regulation of the VSMC phenotype. Indeed, nidogen-2–deficient VSMCs exhibited loss of contractile phenotype in vitro, and compared with wild-type mice, nidogen-2–/– mice showed aggravated post–wire injury neointima formation of carotid arteries. Further bioinformatics analysis, coimmunoprecipitation assays, and luciferase assays revealed that nidogen-2 specifically interacted with Jagged1, a conventional Notch ligand. Nidogen-2 maintained the VSMC contractile phenotype via Jagged1-Notch3 signaling but not Notch1 or Notch2 signaling. Nidogen-2 enhanced Jagged1 and Notch3 interaction and subsequent Notch3 activation. Reciprocally, Jagged1 and Notch3 interaction, signaling activation, and Jagged1-triggered VSMC differentiation were significantly repressed in nidogen-2–deficient VSMCs. In accordance, the suppressive effect of Jagged1 overexpression on neointima formation was attenuated in nidogen-2–/– mice compared with wild-type mice. Conclusions: Nidogen-2 maintains the contractile phenotype of VSMCs through Jagged1-Notch3 signaling in vitro and in vivo. Nidogen-2 is required for Jagged1-Notch3 signaling.