Homeostasis in Adult Vessels

成人血管的稳态

• 批准号: 9386060
• 负责人: M. LUISA IRUELA-ARISPE
• 金额: $ 12.23万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-12-01 至 2018-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9386060
• 关键词: 3' Untranslated Regions; Acceleration; Adult; Anti-inflammatory; Apolipoprotein E; Arteries; Atherosclerosis; Basement membrane; Blood; Blood Vessels; CXCL12 gene; Cardiovascular Diseases; Cell Surface Receptors; Cells; ChIP-seq; Clinical Trials; Collaborations; Conflict (Psychology); Data; Development; Diet; Dietary Fats; Dissociation; Down-Regulation; Endothelial Cells; Endothelium; Event; Exhibits; Exposure to; Gene Targeting; Genes; Genetic; Genomics; Goals; Harvest; High Density Lipoproteins; Homeostasis; Human; In Vitro; Infiltration; Inflammation; Inflammatory; Inflammatory Response; Investigation; Knockout Mice; Lead; Lesion; Letters; Leukocytes; Link; Lipids; Maintenance; Manuscripts; Mediator of activation protein; Messenger RNA; Microarray Analysis; Molecular; Mus; NOTCH1 gene; Oxides; Pathway interactions; Phenotype; Phospholipids; Predisposing Factor; Prevention; Process; Proteins; RNA-Binding Proteins; Regulation; Reporting; Role; Series; Signal Pathway; Signal Transduction; Small Interfering RNA; Surface; TIS11 protein; Testing; Tissues; Transcript; athero susceptible; atheroprotective; base; cancer therapy; cohort; cytokine; design; experimental study; genome wide association study; in vivo; interest; mouse model; notch protein; overexpression; oxidized phosphatidyl choline; pressure; prevent; programs; public health relevance; receptor; recruit; response; senescence; transcription factor; transcriptome sequencing; western diet

 DESCRIPTION (provided by applicant): The long term goal of this line of investigation is to identify the factors that can provide inherent protection to the endothelium during the initial stages of atherosclerosis and inflammation. In this proposal, we will focus on NOTCH1, a cell surface receptor and transcription factor that, based on preliminary data, provides an anti-inflammatory phenotype to quiescent (homeostatic) endothelium. NOTCH1 is constitutively expressed by the adult endothelium of mouse and human vessels in vivo. Reduction of NOTCH1 transcripts in human endothelium in vitro or genetic inactivation of Notch1 in mice triggers an inflammatory response in the absence of any additional insult. Exposure of endothelial cells to Western diet, oxidized phospholipids (Ox- PAPC), as well as inflammatory cytokines, results in a rapid reduction in endogenous NOTCH1 (together with its target genes). Using microarray analyses of endothelial cells from 147 human donors, we observed differences in basal and Ox-PAPC treated levels of NOTCH1 and identified a locus that was associated with the response of endothelial cells to NOTCH1 by Ox-PAPC. This same locus was also associated with HDL levels in a large scale GWAS including 100,000 humans. Studies in this project will test the hypothesis that reduction of NOTCH1 by circulating lipids contributes to the prolonged inflammation typical of atherosclerosis lesions. In fact, reduction of NOTCH1 levels in an athero-susceptible background (Apoe or Ldlr null mice) promotes acceleration of atherosclerosis and results in larger lesions. Furthermore, genetic inactivation of Notch1 in the endothelium of adult mice leads to leukocyte infiltration, detachment and loss of endothelial cells from the intima. In addition and consistent with a role in endothelial homeostasis and suppression of inflammation, using a ChIP approach we found that NOTCH1 regulates tristetraprolin, an RNA binding protein that targets to AREs sequences in the 3'UTR causing destabilization of mRNAs encoding a cohort of inflammatory cytokines. The central hypothesis of this application is that NOTCH1 in the endothelium is essential to maintain an anti-inflammatory interface between blood and tissue. To test this hypothesis, we propose two specific aims: 1. To determine the mechanism(s) that control NOTCH1 expression and function in adult arterial endothelium; and 2. To identify the molecular pathways by which NOTCH1 maintains an anti-inflammatory status in the endothelium of quiescent arteries.

 描述（由申请人提供）：该研究的长期目标是确定在动脉粥样硬化和炎症的初始阶段可以为内皮提供固有保护的因素。在本提案中，我们将重点关注 NOTCH1，一种细胞表面受体和转录因子，根据初步数据，它为静止（稳态）内皮细胞提供抗炎表型。 NOTCH1 在体内由小鼠和人血管的成体内皮细胞组成型表达。在没有任何额外损伤的情况下，体外人内皮细胞中 NOTCH1 转录物的减少或小鼠中 Notch1 的基因失活会引发炎症反应。内皮细胞暴露于西方饮食、氧化磷脂 (Ox-PAPC) 以及炎症细胞因子，会导致内源性 NOTCH1（及其靶基因）迅速减少。通过对 147 名人类供体的内皮细胞进行微阵列分析，我们观察到基础和 Ox-PAPC 处理的 NOTCH1 水平的差异，并确定了与内皮细胞通过 Ox-PAPC 对 NOTCH1 的反应相关的位点。在包括 100,000 人的大规模 GWAS 中，同一基因座也与 HDL 水平相关。该项目的研究将检验以下假设：通过循环脂质减少 NOTCH1 有助于 动脉粥样硬化病变典型的长期炎症。事实上，在动脉粥样硬化易感背景（Apoe 或 Ldlr 缺失小鼠）中，NOTCH1 水平的降低会促进动脉粥样硬化的加速并导致更大的病变。此外，成年小鼠内皮细胞中Notch1基因失活会导致白细胞浸润、内皮细胞脱落和损失 来自内膜。此外，与内皮稳态和炎症抑制中的作用一致，使用 ChIP 方法，我们发现 NOTCH1 调节 tristetraprolin，这是一种 RNA 结合蛋白，靶向 3'UTR 中的 ARE 序列，导致编码一组炎症细胞因子的 mRNA 不稳定。该应用的中心假设是内皮细胞中的 NOTCH1 对于维持血液和组织之间的抗炎界面至关重要。为了检验这一假设，我们提出了两个具体目标： 1. 确定成人动脉内皮细胞中 NOTCH1 表达和功能的控制机制； 2. 确定 NOTCH1 在静止动脉内皮中维持抗炎状态的分子途径。