Vascular Mimetics to Study Inflammation and Atherosclerosis

描述（由申请人提供）：本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 循环中富含甘油三酯的脂蛋白（TGRL），尤其与高饱和脂肪饮食有关。在这笔资助的任期内，我们通过研究从健康、肥胖和高甘油三酯血症受试者血浆中分离出的血单核细胞和餐后TGRL，开发了动脉粥样硬化早期炎症事件的离体模型。通过检查它们与低传代培养的人主动脉内皮细胞 (HAEC) 的相互作用，我们发现急性暴露于 TGRL 会增强细胞因子诱导的 VCAM-1 表达和剪切应力下单核细胞的募集。在表现出高心血管风险（即高餐后 TGRL 和腹部肥胖）的受试者中，我们检测到血液中泡沫单核细胞增加，并将其与整合素激活和 VCAM-1 停滞相关联，如我们的血管模拟剪切流测定中所量化。这些数据提供了量化个体动脉粥样硬化易感性的框架。我们的主要假设是，我们可以描绘出错的代谢途径，并将其与离体测量的内皮和单核细胞炎症生物标志物相关联，以便为评估个体患炎症介导的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体技术，研究细胞和分子尺度的代谢和生物力学反应，以实现三个具体目标：1) 根据 TGRL 的炎症能力，量化高脂膳食后个体的代谢特征。 2) 描述 TGRL 信号通路如何调节主动脉内皮的动脉粥样硬化特征。 3) 使用芯片实验室检测确定与高甘油三酯血症和肥胖相关的最早事件，这些事件会增加单核细胞在致动脉粥样硬化内皮细胞上停滞的倾向。我们的总体目标是开发测定方法，查明来自健康和代谢综合征受试者的膳食脂蛋白发挥促炎作用的程度，从而将平衡从健康脉管系统转变为致动脉粥样硬化脉管系统。

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.