Vascular Mimetics to Study Inflammation and Atherosclerosis

描述(由申请者提供)：这次竞争性更新的重点是动脉粥样硬化，这是西方社会死亡的主要原因。我们的研究和其他研究表明，动脉斑块的形成与内脏肥胖和伴随的高水平密切相关。 循环中的富含甘油三酯的脂蛋白(TGRL)，特别是与高饱和脂肪饮食有关。在这笔赠款的任期内，我们通过研究从健康到肥胖和高甘油三酯血症的受试者的单核细胞和餐后血浆中分离的TGRL，建立了动脉粥样硬化早期炎症事件的体外模型。通过研究它们与低传代培养的人主动脉内皮细胞(HAEC)的相互作用，我们报道了在切应力下，TGRL急性暴露增加细胞因子诱导的VCAM-1的表达和单核细胞的募集。在心血管风险高的受试者(即餐后高TGRL和腹型肥胖)中，我们检测到血液中泡沫单核细胞的增加，并将其与整合素激活和VCAM-1的抑制相关联，正如我们的血管模拟剪切流试验中所定量的那样。这些数据为量化个体的动脉粥样硬化易感性提供了一个框架。我们的主要假设是，我们可以描绘出出错的代谢途径，并将这些途径与体外测量的内皮细胞和单核细胞炎症的生物标志物相关联，以便为评估个人患炎症介导性冠状动脉疾病的风险提供有价值的临床见解。应用系统生物工程方法，利用实时成像和新型微流控技术，在细胞和分子水平上研究代谢和生物力学响应，有三个特定目标：1)根据TGRL的炎症能力来量化个人在高脂饮食后的代谢情况。2)阐明TGRL信号通路对动脉内皮细胞致动脉粥样硬化的调节作用。3)使用芯片实验室分析确定与高甘油三酯血症和肥胖有关的最早事件，这些事件增加了单核细胞在致动脉粥样硬化的内皮细胞上停滞的倾向。我们的总体目标是开发准确的分析方法，以确定来自健康和代谢综合征受试者的饮食脂蛋白在多大程度上发挥促炎作用，将平衡从健康血管转变为动脉粥样硬化。

DESCRIPTION (provided by applicant): This competitive renewal is focused on atherosclerosis, the primary cause of mortality in Western society. Our studies and others reveal that arterial plaque formation closely correlates with visceral obesity and concomitant high levels of circulating triglyceride rich lipoproteins (TGRL), especially associated with diets high in saturated fat. Over the tenure of this grant, we have developed an ex-vivo model of early inflammatory events of atherogenesis by studying blood monocytes and TGRL isolated postprandial from plasma of subjects ranging from healthy to obese and hypertriglyceridemic. By examining their interaction with low passage cultured human aortic endothelial cells (HAEC), we have reported that acute exposure to TGRL augmented cytokine- induced VCAM-1 expression and monocyte recruitment under shear stress. In subjects exhibiting high cardiovascular risk (i.e. high postprandial TGRL and abdominal obesity) we detected increased foamy monocytes in blood and correlated this with integrin activation and arrest to VCAM-1, as quantified in our vascular mimetic shear flow assay. These data provide a framework for quantifying an individual's atherogenic susceptibility. Our primary hypothesis is that we can delineate the metabolic pathways that go awry and correlate these with biomarkers of endothelial and monocyte inflammation measured ex vivo in order to provide valuable clinical insight in assessing an individual's risk for developing inflammation-mediated coronary artery disease. A systems bioengineering approach is applied to study metabolic and biomechanical responses at the cell and molecular scale using real time imaging and novel microfluidic technology in three specific aims: 1) Quantify an individual's metabolic profile following a high-fat meal in terms of the inflammatory capacity of their TGRL. 2) Delineate how TGRL signaling pathways regulate an atherogenic profile in aortic endothelium. 3) Identify the earliest events linked to hypertriglyceridemia and obesity that increase monocyte proclivity to arrest on atherogenic endothelium using a lab-on-a-chip assay. Our overall goal is to develop assays that pinpoint the extent to which dietary lipoproteins from healthy and metabolic syndrome subjects are exerting pro- inflammatory effects that shift the balance from healthy to atherogenic vasculature.