OSTEOPONTIN AND OXIDATIVE STRESS IN ATHEROGENESIS

骨桥蛋白和动脉粥样硬化形成中的氧化应激

• 批准号: 6638536
• 负责人: Kenneth S. Ramos
• 金额: $ 25.51万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2005-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6638536
• 关键词: AP1 protein; DNA footprinting; atherosclerosis; cell proliferation; epidermal growth factor; extracellular matrix; flow cytometry; focal adhesion kinase; gel mobility shift assay; genetic transcription; in situ hybridization; integrins; laboratory rat; molecular pathology; nuclear factor kappa beta; nuclear runoff assay; osteopontin; oxidative stress; phosphorylation; polymerase chain reaction; vascular smooth muscle

DESCRIPTION (Verbatim from the application): Atherosclerosis is a complex disease process characterized by uncontrolled proliferation of vascular smooth muscle cells (vSMCs) and alterations in cell/cell and cell/matrix interactions. Using a chemical model of oxidative injury to the blood vessel wall in vivo, we have recently shown that modulation of vSMCs to proliferative phenotypes involves overexpression of osteopontin (OPN), an integrin ligand, and alterations in integrin-coupled mitogenic signaling. Based on these observations we hypothesized that transition of vSMCs to proliferative phenotypes is mediated by alterations in OPN expression and extracellular matrix (ECM) signaling. To test this hypothesis, studies are proposed in Aim l to define the molecular bases of OPN upregulation as a function of atherogenic status in vSMCs induced by oxidative stress. Nuclear run-on, promoter mutation/deletion analysis, electrophoretic mobility shift assays, mRNA stability measurements, and DNA footprinting will be completed to define the transcriptional and post-transcriptional component of the OPN response. Primary emphasis will be given to NF-KB- and AP-1 coupled signal transduction cascades in the regulation of ECM-regulated gene expression. As part of the second aim, studies will be conducted to characterize the integrin profiles of modified SMCs relative to control counterparts, and to define the molecular basis of signaling interference. Fluorescence activated cell sorting, northern analysis, reverse transcription polymerase chain reaction (PCR), and in situ hybridization will be conducted to identify specific integrins targeted by oxidative injury. The final series of experiments will characterize the interaction between OPN and EGF signaling pathways as a function of phenotypic status. This aim will involve study of EGF receptor phosphorylation, focal adhesion kinase measurements, and downstream EGF signal transduction cascades to identify signals that lead to convergence of the EGF and ECM signaling during atherogenesis. Collectively, the proposed studies will serve to elucidate molecular mechanisms of integrin-related signaling during the induction of atherogenic vascular SMC phenotypes by oxidative injury. These studies are important in view of the increasing scrutiny of the role of oxidative injury in the onset and progression of atherogenesis.

描述（逐字化的应用程序）：动脉粥样硬化是一个复杂的 疾病过程的特征是血管光滑的不受控制 肌肉细胞（VSMC）和细胞/细胞和细胞/基质相互作用的改变。 在体内使用对血管壁的氧化损伤的化学模型，我们 最近表明，VSMC对增殖表型的调节 涉及骨桥蛋白（OPN），整联蛋白配体和 整联蛋白偶联有丝分裂信号传导的改变。基于这些 观察结果我们假设VSMC向增殖的过渡 表型是由OPN表达和细胞外改变的介导的 矩阵（ECM）信号传导。为了检验这一假设，提出了AIM L的研究 定义OPN上调的分子碱基作为动脉粥样硬化的函数 由氧化应激引起的VSMC状态。核跑步，发起人 突变/缺失分析，电泳迁移率转移测定，mRNA 稳定性测量和DNA足迹将完成以定义 OPN响应的转录和转录后成分。基本的 将重点放在NF-KB和AP-1耦合信号转导级联 在调节ECM调节的基因表达中。作为第二个目标的一部分， 将进行研究以表征经过修饰的整联蛋白轮廓 SMC相对于控制对应物，并定义 信号干扰。荧光激活的细胞分选，北部分析， 逆转录聚合酶链反应（PCR）和原位 将进行杂交以确定由 氧化损伤。最后一系列实验将表征 OPN和EGF信号通路之间的相互作用随表型的函数 地位。该目标将涉及研究EGF受体磷酸化，局灶性 粘附激酶测量和下游EGF信号转导级联 确定导致EGF和ECM信号收敛的信号 在动脉粥样硬化期间。拟议的研究总共有助于 阐明整联蛋白相关信号传导的分子机制 通过氧化损伤诱导动脉粥样硬化血管SMC表型。这些 鉴于对越来越多的审查的作用，研究很重要 动脉粥样硬化的发作和进展中的氧化损伤。