层流剪应力在维持血管内皮稳态中发挥重要作用。我们前期研究发现层流状态下内皮中的AMPK可使Nucleolin S328磷酸化，而已知Nucleolin在调节转录及RNA生成方面具有重要的生理意义，因此我们推测层流剪应力激活AMPK/Nucleolin是维持内皮细胞稳态的关键分子事件。本项目拟采用表观遗传学、转录组学及系统生物学等手段探讨层流剪应力-AMPK-Nucleolin影响内皮细胞功能的分子机制，采用CRISPR/Cas9技术实现Nucleolin S328D（激活型）、S328A（失活型）在体基因定点突变，揭示Nucleolin S328磷酸化对内皮的保护作用及对动脉粥样硬化形成的影响，探讨Nucleolin磷酸化作为动脉粥样硬化干预靶点的可能性。本项目将进一步阐明血流剪应力影响内皮细胞功能的核心作用，并在此基础上明确AMPK-Nucleolin动态网络在血管生物学中的转化意义。

The atheroprotective flow maintains the homeostasis of endothelial cells in the vasculature. We have previously found that atheroprotective flow causes AMPK phosphorylation of Nucleolin at Ser-328. Given the multifaceted role of Nucleolin in regulating transcription and RNA biogenesis, we propose to study the role of atheroprotective flow activation of the AMPK-Nucleolin axis in regulating endothelium in vitro and in vivo. We will use multi-level approaches including epigenetics, transcriptional genomics, and systems biology to investigate the mechanotransduction and transcriptional regulatory network involved in the AMPK-Nucleolin axis in endothelial cells responding to the atheroprotective flow. To explore the functional consequence of the flow regulation of AMPK-Nucleolin in endothelial function in vivo, we will use CRISPR/Cas9 technology to create mice harboring Nucleolin S328D (phosphomimetic) or S328A (dephosphomimetic) mutant. These mice will be crossed with ApoE-null strain for the investigation of atherosclerosis. With these approaches at the systems level, this research project will decipher the role of atheroprotective flow in maintaining a functional endothelium via regulating the AMPK-Nucleolin axis.