Department of Physiology and Pathophysiology (C.M., Z.M., Y.J., N.X., G.Z., B.M., F.Y., J.S., Y.F., W.K.), School of Basic Medical Sciences, Peking University Health Science Center, Beijing, China.
Key Laboratory of Molecular Cardiovascular Science, Ministry of Education, Beijing, China (C.M., Z.M., Y.J., N.X., G.Z., B.M., F.Y., Y.F., W.K.).
Circulation. 2021 Oct 12;144(15):1244-1261. doi: 10.1161/CIRCULATIONAHA.120.053361. Epub 2021 Jul 28.
How the extracellular matrix (ECM) microenvironment modulates the contractile phenotype of vascular smooth muscle cells (VSMCs) and confers vascular homeostasis remains elusive.
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.
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.
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.
细胞外基质 (ECM) 微环境如何调节血管平滑肌细胞 (VSMC) 的收缩表型并赋予血管稳态仍然难以捉摸。
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，我们确定了基底膜糖蛋白 nidogen-2 是维持血管平滑肌细胞特性的关键 ECM 蛋白。
我们通过使用基因本体论注释结合文献搜索收集了与 VSMC 表型相关的基因数据集。对 ECM 蛋白基因和 VSMC 表型相关基因数据集之间的蛋白质-蛋白质相互作用的计算分析揭示了候选基因 nidogen-2，这是一种参与调节 VSMC 表型的基底膜糖蛋白。事实上，与野生型小鼠相比，nidogen-2 缺陷型 VSMC 在体外表现出收缩表型丧失，nidogen-2 小鼠表现出颈动脉损伤后新生内膜形成加重。进一步的生物信息学分析、共免疫沉淀测定和荧光素酶测定显示，nidogen-2 特异性与 Jagged1 相互作用，Jagged1 是一种传统的 Notch 配体。nidogen-2 通过 Jagged1-Notch3 信号维持 VSMC 收缩表型，而不是 Notch1 或 Notch2 信号。nidogen-2 增强了 Jagged1 和 Notch3 的相互作用以及随后的 Notch3 激活。相反，在 nidogen-2 缺陷型 VSMC 中，Jagged1 和 Notch3 相互作用、信号激活以及 Jagged1 触发的 VSMC 分化显著受到抑制。相应地，与野生型小鼠相比，Jagged1 过表达对新生内膜形成的抑制作用在 nidogen-2 小鼠中减弱。
nidogen-2 通过体外和体内的 Jagged1-Notch3 信号维持 VSMC 的收缩表型。nidogen-2 是 Jagged1-Notch3 信号所必需的。
