MST1 Suppresses Disturbed Flow Induced Atherosclerosis
MST1 Suppresses Disturbed Flow Induced Atherosclerosis
复制标题
DOI：
10.1161/circresaha.122.321322
复制
发表时间：
2022-10-14
影响因子：
20.1
通讯作者：
Ai, Ding
中科院分区：
文献类型：
--
作者：
Quan, Meixi;Lv, Huizhen;Ai, Ding
背景：动脉粥样硬化主要发生在血流受到干扰的动脉分支点。 MST1（哺乳动物不育 20 样激酶 1）是机械敏感 Hippo 通路中的主要激酶，如何调节血流紊乱诱导的内皮细胞 (EC) 激活和动脉粥样硬化仍不清楚。方法：为了评估 MST1 在体内的作用，将 ApoE(-/-) 背景上 EC 特异性 Mst1 缺陷 (Mst1(iECKO)ApoE(-/-)) 的小鼠用于通过颈动脉结扎产生的动脉粥样硬化模型。使用质谱、免疫沉淀、邻近连接测定和染料摄取测定来鉴定MST1的功能底物。在体外进行的实验中，人脐静脉内皮细胞和人主动脉内皮细胞受到模拟扰动流动的振荡剪切应力。结果：我们发现，在人和小鼠动脉和 EC 的振荡剪切应力暴露区域中，内皮 MST1 的磷酸化受到显着抑制。异位慢病毒介导的野生型 MST1 过表达，而非 MST1 激酶缺陷型突变体，可逆转 ApoE(-/-) 背景上 EC 特异性 Mst1 缺陷中扰动的 EC 激活和动脉粥样硬化 (Mst1(iECKO)ApoE(-/-))。振荡剪切应力抑制 MST1 会导致 Cx43（连接蛋白 43）Ser255 磷酸化减少、Cx43 半通道开放、EC 激活和动脉粥样硬化，而这些都被 Cx43 半通道抑制肽 TAT-GAP19 阻断。质谱研究发现，Filamin B 促进了 Cx43 易位至脂筏，以进一步打开半通道。最后，慢病毒介导的Cx43（S255）突变体过表达为谷氨酸以模拟磷酸化，减弱了受干扰的血流诱导的EC激活，从而抑制ApoE（-/-）和Mst1（iECKO）ApoE（-/-）小鼠中的动脉粥样硬化形成。结论：我们的研究表明，抑制MST1-Cx43轴是振荡剪切应力诱导的内皮功能障碍和动脉粥样硬化的重要驱动因素，这为动脉粥样硬化的治疗提供了新的治疗靶点。
Background: Atherosclerosis occurs mainly at arterial branching points exposed to disturbed blood flow. How MST1 (mammalian sterile 20-like kinase 1), the primary kinase in the mechanosensitive Hippo pathway modulates disturbed flow induced endothelial cells (ECs) activation and atherosclerosis remains unclear. Methods: To assess the role of MST1 in vivo, mice with EC-specific Mst1 deficiency on ApoE(-/-) background (Mst1(iECKO)ApoE(-/-)) were used in an atherosclerosis model generated by carotid artery ligation. Mass spectrometry, immunoprecipitation, proximity ligation assay, and dye uptake assay were used to identify the functional substrate of MST1. Human umbilical vein endothelial cells and human aortic endothelial cells were subjected to oscillatory shear stress that mimic disturbed flow in experiments conducted in vitro. Results: We found that the phosphorylation of endothelial MST1 was significantly inhibited in oscillatory shear stress-exposed regions of human and mouse arteries and ECs. Ectopic lenti-mediated overexpression of wild-type MST1, but not a kinase-deficient mutant of MST1, reversed disturbed flow-caused EC activation and atherosclerosis in EC-specific Mst1 deficiency on ApoE(-/-) background (Mst1(iECKO)ApoE(-/-)). Inhibition of MST1 by oscillatory shear stress led to reduced phosphorylation of Cx43 (connexin 43) at Ser255, the Cx43 hemichannel open, EC activation, and atherosclerosis, which were blocked by TAT-GAP19, a Cx43 hemichannel inhibitory peptide. Mass spectrometry studies identified that Filamin B fueled the translocation of Cx43 to lipid rafts for further hemichannel open. Finally, lenti-mediated overexpression of the Cx43(S255) mutant into glutamate to mimic phosphorylation blunted disturbed flow-induced EC activation, thereby inhibiting the atherogenesis in both ApoE(-/-) and Mst1 (iECKO)ApoE(-/-) mice. Conclusions: Our study reveals that inhibition of the MST1-Cx43 axis is an essential driver of oscillatory shear stress-induced endothelial dysfunction and atherosclerosis, which provides a new therapeutic target for the treatment of atherosclerosis.