Cytokine signaling in activation of the endothelium

内皮激活中的细胞因子信号传导

• 批准号: 7659839
• 负责人: PAUL E DICORLETO
• 金额: $ 43.29万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-15 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7659839
• 关键词: Adhesions; Adult; Affect; Anti-Inflammatory Agents; Anti-inflammatory; Apolipoprotein E; Arginine; Arterial Fatty Streak; Atherosclerosis; Binding; Biochemical; Blood Vessels; Cell Nucleus; Cell Surface Receptors; Cell surface; Cells; Coronary Arteriosclerosis; Cytokine Activation; Cytokine Signaling; Data; Development; Disease; Dissociation; E-Selectin; Endothelial Cells; Endothelium; Event; Gene Activation; Gene Expression; Gene Targeting; Genes; Genetically Engineered Mouse; HOXA9 gene; HOXA9 protein; Human; In Vitro; Inflammation; Inflammatory; Inflammatory Response; Instruction; Intercellular adhesion molecule 1; Knockout Mice; Leukocyte Adhesion Molecules; Leukocyte Rolling; Leukocyte-Adhesion Receptors; Leukocytes; Mediating; Mediator of activation protein; Modeling; Molecular; Molecular Biology; Mus; Play; Post-Translational Protein Processing; Process; Proteins; Proteomics; Qualifying; Regulation; Rheumatoid Arthritis; Role; Signal Pathway; Signal Transduction; Site; Site-Directed Mutagenesis; Small Interfering RNA; Stem cells; Stimulus; System; TNF gene; TNFRSF1B gene; Technology; Testing; Therapeutic; Tibial Arteries; Tissues; Transferase; Tumor Necrosis Factor Receptor; Tumor Necrosis Factor-alpha; Tumor Necrosis Factors; Vascular Cell Adhesion Molecule-1; Vascular Endothelial Cell; angiogenesis; base; chromatin immunoprecipitation; chromatin remodeling; cremaster muscle; cytokine; deletion analysis; experience; human MPP1 protein; human tissue; in vivo; intravital microscopy; mouse model; novel; promoter; receptor; response; transcription factor; transcriptional coactivator p75; tumor necrosis factor receptor 1A

The central objective of this Project is to define signaling mediators and transcription factors of critical importance in cytokine activation of vascular endothelial cells (EC). Our specific focus is on TNF-a receptor-ll (p75) and the homeobox protein HOXA9. We have recently shown that both molecules are required for cytokine induction of the EC- leukocyte adhesion molecule E-selectin, as well as other activation genes. We showed that TNF-stimulated leukocyte firm adhesion to EC was dramatically reduced in p75-null mice, but not affected in p55 (the other TNF receptor)-null mice. Both receptors participated in TNF-induced leukocyte rolling and transmigration. Furthermore, both p75 and p55 were required for TNF induction of E-selectin and VCAM-1; whereas, p75 was sufficient for ICAM-1 induction. We also demonstrated that p75-deficiency in apoE-null mice reduced atherosclerotic lesion development. Based on these findings, we hypothesize that p75 activation in EC triggers intra-cellular signaling pathways, not induced by p55, that are essential for TNF-induced pro-inflammatory gene expression in EC. We have also recently shown that the transcription factor HOXA9 is required for TNF induction of E-selectin in EC due to its transient binding to a newly identified Abd-B-like site in the promoter. The TNF signal that leads to association of HOXA9 with the E-selectin pro- moter is triggered by p75 and not p55. We hypothesize that HOXA9 plays a novel role in the activation of EC in res- ponse to inflammatory stimuli. We propose to test our hypotheses by pursuing three aims. The first is to identify TNF- induced, p75-specific signaling pathways and novel p75-induced genes in EC using TNF-treated mouse EC isolated from p55-/-, p75-/- and p55-p75-double null mice. We will also identify TNF-induced binding partners of p75 and determine structural domains in p75 responsible for induced gene expression in EC. The second aim is to elucidate the molecular mechanism of HOXA9-mediated E-selectin expression and to identify novel target genes for HOXA9 in EC. We will pursue post-translational modifications of HOXA9 and its possible binding to other obligate transcription factors for E-selectin induction. Gene and promoter array studies will be used to identify novel target genes of HOXA9. In the third aim we will characterize the role of the HOXA9-binding partner protein arginine methyl transferase 5 (PRMT5) in activated EC using biochemical, siRNA-based and pharmacological approaches. We also plan to identify proteins methylated by PRMT5 that may qualify for novel mediators of EC activation. RELEVANCE (See instructions): EC, which line all blood vessels in the body, play a key role in inflammation - a process that underlies many diseases, including atherosclerosis. Our studies will reveal new molecular players in the regulation of inflammation. These players will represent novel targets for the development of anti-inflammatory therapeutics that may be used to treat such diseases as coronary artery disease and rheumatoid arthritis.

该项目的核心目的是定义信号传导介质和转录因子至关重要的转录因子 血管内皮细胞的细胞因子激活（EC）。我们的具体重点是TNF-A受体-LL（p75）和 同源蛋白Hoxa9。我们最近表明，两个分子都是EC-的细胞因子诱导所必需的 白细胞粘附分子E-选择素以及其他激活基因。我们证明了TNF刺激的白细胞 在P75-NULL小鼠中，对EC的牢固粘附大大降低，但在p55（其他TNF受体）中不受影响 老鼠。两个受体都参与TNF诱导的白细胞滚动和移民。此外，P75和P55均 TNF诱导E-选择蛋白和VCAM-1是必需的；而P75足以用于ICAM-1诱导。我们也是 证明APOE-NULL小鼠的P75缺乏症会降低动脉粥样硬化病变的发育。基于这些 调查结果，我们假设EC中的p75激活触发了细胞内信号传导途径，而不是由p55诱导的， 对于TNF诱导的EC中的促炎基因表达至关重要。我们最近还表明了 转录因子HOXA9是TNF在EC中诱导E-选择蛋白的瞬时结合所必需的 在启动子中鉴定出类似ABD-B的位点。 TNF信号导致HOXA9与E-选择蛋白促蛋白的关联 电动机由p75而不是p55触发。我们假设Hoxa9在EC的激活中起着新颖的作用 对炎症刺激的贡献。我们建议通过追求三个目标来检验我们的假设。首先是确定TNF- 使用TNF处理的小鼠EC分离的诱导的P75特异性信号通路和EC中的新型P75诱导的基因 来自p55 - / - ，p75 - / - 和p55-p75双零小鼠。我们还将确定p75和 确定负责EC中诱导基因表达的p75中的结构结构域。第二个目的是阐明 HOXA9介导的E-选择素表达的分子机制，并在EC中鉴定Hoxa9的新靶基因。 我们将追求HOXA9的翻译后修改及其可能与其他义务转录因子的结合 用于电子选择蛋白诱导。基因和启动子阵列研究将用于鉴定HOXA9的新靶基因。在 第三目的我们将表征HOXA9结合伴侣蛋白精氨酸转移酶5（PRMT5）的作用 在使用生化，基于siRNA的药理方法的激活的EC中。我们还计划识别蛋白质 由PRMT5甲基化，可能有资格获得EC激活的新型介体。 相关性（请参阅说明）： EC在体内的所有血管中排成一列，在炎症中起着关键作用 - 这一过程是许多疾病的基础 包括动脉粥样硬化。我们的研究将揭示炎症调节中的新分子参与者。这些球员 将代表开发抗炎治疗剂的新目标，这些疗法可用于治疗这种治疗 作为冠状动脉疾病和类风湿关节炎的疾病。