Nucleolin Regulation of miRome by Shear Stress

动脉树中动脉硬化病变的局灶性分布可归因于血管的暴露。 血管分支点和弯曲处的内皮细胞(ECs)到动脉粥样硬化的振荡切变(OS)，在 与血管直部的抗动脉粥样硬化的脉动切变(PS)形成对比。这两种类型的流 模式诱导不同的miRNAs(MiRs)表达谱，从而差异地调节靶基因的表达 基因，导致不同的功能结果。核仁素(NCL)是一种多功能蛋白质，在蛋白质中起重要作用。 通过表观遗传、转录、转录后和翻译调节细胞功能 规章制度。我们认为NCL通过其相互作用来调控miRoman(miR转录和成熟)。 DNA(通过g-四链序列)和RNA(通过UCCCGA共有序列)。我们的初步结果 已经证明PS诱导NCL的磷酸化，从而导致NCL与 MiR23b/27b启动子/增强子区域促进miR转录。另一方面，在操作系统下，非 磷酸化的NCL与miR93/miR484和DROSHA结合，促进miR成熟。这些新奇的发现 导致了这一更新建议的假说：NCL受到动脉粥样硬化保护和 动脉粥样硬化蛋白在转录和成熟水平上的流动模式，从而发挥作用 在健康和疾病中对血管动态平衡起着重要作用。为了检验这一假设，我们提出了 以下四个具体目标：1)研究NCL PS-vs.OS-调控的分子机制 在miR转录组中，2)确定NCL-PS与OS-调控的分子机制。 调制miR成熟，3)将NCL的miRoman多层规则与功能 在PS和OS下使用系统方法的后果，4)验证NCL在监管中的作用 体内的血管功能。在这项拟议的研究中，我们将使用体外内皮细胞生物学、 活体小鼠模型，在电子网络构建/数据挖掘中，以及来自患者的临床样本 心血管疾病，以破译NCL的血流调节机制--MICROMA的调节。这个 这些发现将导致对NCL在调节健康和疾病中的血管功能中的作用的新的理解 通过MIR法规。 好了！

The focal distribution of atherosclerotic lesions in the arterial tree is attributable to the exposure of the vascular endothelial cells (ECs) to the atheroprone oscillatory shear (OS) at vessel branch points and curvatures, in contrast to the atheroprotective pulsatile shear (PS) at straight parts of the vessel. These two types of flow patterns induce distinct expression profiles of miRNAs (miRs) that differentially regulate the expression of target geness, resulting in distinct functional outcomes. Nucleolin (NCL) is a multi-functional protein that plays critical roles in regulating cell functions through epigenetic, transcriptional, post-transcriptional, and translational regulations. We propose that NCL regulates miRome (miR transcription and maturation) through its interactions with DNA (via g-quadruplex sequences) and RNA (via UCCCGA consensus sequence). Our preliminary results have demonstrated that PS induces the phosphorylation of NCL that leads to an increased binding of NCL to the promoter/enhancer regions of miR23b/27b to increase miR transcription. On the other hand, under OS, the non- phosphorylated NCL binds to miR93/miR484 and Drosha to facilitate miR maturation. These novel findings have led to the hypothesis of this renewal proposal: NCL is differentially regulated by atheroprotective and atheroprone flow patterns to modulate the miRome at transcriptional and maturation levels, thus playing an important role in vascular homeostasis in health and disease. To test this hypothesis, we propose the following four Specific Aims: 1) to examine the molecular mechanisms underlying PS- vs. OS-regulation of NCL in miR transcriptome, 2) to determine the molecular mechanisms underlying PS- vs. OS-regulation of NCL- modulated miR maturation, 3) to integrate the multi-layer regulations of miRome by NCL and the functional consequences under PS and OS with the use of systems approaches, 4) to validate the role of NCL in regulating vascular function in vivo. In the proposed research, we will use a combination of in vitro endothelial cell biology, in vivo mouse models, in silico network construction/data mining, and clinical samples derived from patients with cardiovascular diseases to decipher the mechanisms of flow modulation of NCL-regulation of miRome. The findings will result in novel understanding of the role of NCL in regulating vascular functions in health and disease via miR regulations. !