Engineering an Atherosclerosis-Resistant Endothelium

描述（由申请人提供）：动脉粥样硬化发生在血流模式偏离正常层流模式的血管区域。致动脉粥样硬化的血流动力学激活内皮细胞募集单核细胞并降低屏障功能，这在存在全身性危险因素的情况下导致动脉粥样硬化。最近的数据表明，内皮下细胞外基质的组成在这一过程中起着重要的作用。流体流动激活内皮中的NF-κ B、PAK和JNK！细胞依赖于纤连蛋白（FN）或纤维蛋白原，但不依赖于基底膜蛋白，这是由于结合这些细胞外基质蛋白的整合素的差异信号传导。此外，FN在体内易发生动脉粥样硬化的部位先于其他疾病指征，表明FN的沉积可能有助于体内动脉粥样硬化的发生。我们的体外研究表明，FN基质可以被修饰以抑制NF-κ B和JNK，这提示了预防或治疗动脉粥样硬化的可能方法。该项目将由弗吉尼亚大学的一个研究小组执行，该小组由以下人员组成：Martin Schwartz博士，他从事机械转导和整合素信号传导; Brian黑尔姆克，一位从事机械转导和图像分析的生物工程师; Brett Blackman，一位内皮生物学和测量及建模流动模式的生物工程师专家;和伊恩·萨伦博克，一位心脏病学家和动脉粥样硬化动物模型和免疫组织化学专家。为了利用上述发现所创造的机会，我们将使用血管系统中动脉粥样硬化倾向部位的流动和应变模式来研究致动脉粥样硬化力的体外效应。这些特征将被用来阐明在动脉粥样硬化易感区域引起动脉粥样硬化途径持续激活的信号传导机制。这些信息将被用于工程转染的内皮细胞系，不激活这些途径。为了测试体内结果的相关性，构建在内皮中表达类似构建体的转基因小鼠，并与载脂蛋白E+小鼠杂交，以评价它们抑制动脉粥样硬化的能力。总之，这些研究将提供深入了解动脉粥样硬化的机制和测试新的治疗策略的治疗。

DESCRIPTION (provided by applicant): Atherosclerosis occurs at regions of the vasculature where flow patterns depart from the normal laminar mode. Atherogenic hemodynamic forces activate the endothelium to recruit monocytes and decrease barrier unction, which in the presence of systemic risk factors leads to atherosclerosis. Recent data show that the composition of the subendothelial extracellular matrix plays an important role in this process. Fluid flow activates NF-KB, PAK and JNK in endothelia! cells on fibronectin (FN) or fibrinogen but not on basement membrane proteins due to differential signaling by the integrins that bind these extracellular matrix proteins. Furthermore, FN is found at atherosclerosis-prone sites in vivo prior to other indications of disease, suggesting that deposition of FN may contribute to initiation of atherosclerosis in vivo. Our in vitro studies have shown that FN matrix can be modified to suppress NF-KB and JNK, suggesting a possible method to Drevent or treat atherosclerosis. The project will be performed by a team of researchers at the University of Virginia consisting of Drs. Martin Schwartz, who works on mechanotransduction and integrin signaling; Brian Helmke, a bioengineer who works on mechanotransduction and image analysis; Brett Blackman, a bioengineer expert in endothelial biology and measuring and modeling flow patterns; and Ian Sarembock, a cardiologist and expert in animal models of atherosclerosis and immunohistochemistry. To take advantage of the opportunity created by the above findings, we will use flow and strain patterns from an atherosclerosis- prone site in the vasculature to study effects of atherogenic forces in vitro. These profiles will be used to elucidate mechanisms of signaling that give rise to sustained activation of atherogenic pathways in athero- prone regions. This information will be used to engineer transfected endothelial cell lines that do not activate these pathways. To test the relevance of the results in in vivo, transgenic mice expressing similar constructs in the endothelium will be constructed and crossed with apolipoproteinE"'" mice to evaluate their ability to suppress atherosclerosis. Together, these studies will provide insight into mechanisms of atherogenesis and test novel therapeutic strategies for treatment.