Regulation of Endothelial Cells by the OX-Papc Network

我们的研究表明，在体内积累的磷脂氧化产物(Ox-PAPC) 动脉粥样硬化病变是血管内皮细胞功能的重要调节因子，影响血管内皮细胞的mRNA水平。 1000个涉及炎症、固醇调节、凝血、氧化应激、细胞周期、血管生成、 氧化还原调节和未折叠的蛋白质反应。Ox-PAPC及其受体的炎症反应 成分脂质PEIPC与IPS和TNF显著不同，导致慢性 上调单核细胞与内皮细胞的相互作用。拟议研究的一个主要目标是确定 Ox-PAPC/PEIPC网络的关键调节者使用细胞生物学和生物信息学方法。在AIM 1我们将使用细胞生物学的方法来测试Ox激活的基本信号机制的三个方面： PAPC和PEIPC。我们将：定义Ox-PAPC受体复合体的其他成分；确定如何 OX-PAPC和PEIPC改变细胞氧化还原平衡以控制基因表达，并确定 PEIPC与蛋白质的共价结合在激活过程中起着重要作用。在目标2中，我们将使用一种综合遗传学 在文本级别定义整体网络的方法，利用共同基因的概念 群体中的变异可用于将表达阵列数据组织成生物相关的模块。 我们将使用全基因组关联来绘制导致网络中常见变异的基因 将数据与正交蛋白质组和功能数据集相结合。目标1和目标2还将包括 通过使用siRNA和在某些情况下过度表达来验证重要的调控因子。在《目标3》中，我们将 确定内皮细胞是否表达三种重要的网络调节因子(SREBP、STAT3和HO-1) 在小鼠动脉粥样硬化中起重要作用。在这些研究中，我们将使用低密度脂蛋白受体缺失的小鼠 血管内皮细胞特异性敲除这些蛋白。此外，人体损伤的炎症性区域将 检查这些分子的表达和激活，以及我们发现的其他调节网络的分子。 总之，这些研究将确定控制动脉粥样硬化的潜在内皮靶点。

Our studies have demonstrated that phospholipid oxidation products (Ox-PAPC) which accumulate in atherosclerotic lesions are important regulators of endothelial cell function, affecting mRNA levels of over 1000 genes involving inflammation, sterol regulation, coagulation, oxidative stress, cell cycle, angiogenesis, redox regulation and the unfolded protein response. The inflammatory response to Ox-PAPC and to its component lipid PEIPC was shown to differ significantly from those of IPS and TNF, leading to a chronic upregulation of monocyte-endothelial interactions. A major goal of the proposed studies is to identify the pivotal regulators of the Ox-PAPC/PEIPC network using cell biology and bioinformatics approaches. In Aim 1 we will use a cell biology approach to test three aspects of the basic signaling mechanism activated by Ox- PAPC and PEIPC. We will: define additional components of the Ox-PAPC receptor complex; determine how Ox-PAPC and PEIPC alter the cellular redox balance to contol gene expression; and determine whether covalent binding of PEIPC to proteins is important in activation. In Aim 2, we will use an integrative genetics approach to define the overall network at the transcript level, leveraging the concept that common genetic variations in the population can be used to organize expression array data into biologically relevant modules. We will map the genes contributing to common variation in the network using genome-wide association and integrate the data with orthogonal proteomic and functional datasets. Aims 1 and 2 will also include validation of important regulators by use of siRNA and in some cases overexpression. In Aim 3 we will determine whether endothelial expression of three important network regulators (SREBP, STAT3 and HO-1) plays an important role in atherosclerosis in mice. For these studies, we will employ LDL receptor null mice with endothelial specific knockout of these proteins. In addition, inflammatory areas of human lesions will be examined for expression and activation of these molecules and others we find to regulate the network. Together, these studies will identify potential endothelial targets for the control of atherosclerosis.