Engineering an Atherosclerosis-Resistant Endothelium

描述（由申请人提供）：动脉粥样硬化发生在血管流动模式偏离正常层流模式的区域。致动脉粥样硬化的血流动力学力激活内皮细胞募集单核细胞，降低屏障功能，在系统性危险因素存在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作用。流体流动激活内皮细胞NF-KB、PAK和JNK ！细胞在纤维连接蛋白（FN）或纤维蛋白原上，但不在基底膜蛋白上，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在体内动脉粥样硬化易发部位被发现，比其他疾病适应症更早，这表明FN的沉积可能有助于体内动脉粥样硬化的开始。我们的体外研究表明，FN基质可以被修饰以抑制NF-KB和JNK，这可能是预防或治疗动脉粥样硬化的一种方法。该项目将由弗吉尼亚大学的一组研究人员执行。研究机械转导和整合素信号的Martin Schwartz；从事机械转导和图像分析工作的生物工程师布莱恩·赫尔姆克；布雷特·布莱克曼（Brett Blackman）是内皮生物学和测量和建模血流模式的生物工程师专家；以及心脏病专家、动脉粥样硬化动物模型和免疫组织化学专家Ian Sarembock。为了利用上述发现带来的机会，我们将利用血管动脉粥样硬化易发部位的血流和应变模式，在体外研究动脉粥样硬化力的影响。这些资料将用于阐明在动脉粥样硬化易发区域引起动脉粥样硬化途径持续激活的信号传导机制。这些信息将用于工程转染内皮细胞系，不激活这些途径。为了验证实验结果在体内的相关性，我们构建了在内皮中表达类似结构的转基因小鼠，并将其与载脂蛋白“’”小鼠杂交，以评估其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形成的机制，并测试新的治疗策略。

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.