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，动脉斑块形成与内脏肥胖密切相关， 循环甘油三酯丰富的脂蛋白（TGRL），特别是与高饱和脂肪的饮食有关。在该资助的任期内，我们通过研究从健康到肥胖和高脂血症的受试者的血浆中分离的血液单核细胞和餐后TGRL，开发了动脉粥样硬化形成的早期炎症事件的离体模型。通过检测它们与低传代培养的人主动脉内皮细胞（HAEC）的相互作用，我们报道了在剪切应力下急性暴露于TGRL增强了细胞因子诱导的VCAM-1表达和单核细胞募集。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状单核细胞增加，并将其与整合素活化和VCAM-1阻滞相关，如我们的血管模拟剪切流测定中所定量的。这些数据提供了一个框架，量化个人的动脉粥样硬化易感性。我们的主要假设是，我们可以描绘出差错的代谢途径，并将其与离体测量的内皮细胞和单核细胞炎症的生物标志物相关联，以便在评估个体发生炎症介导的冠状动脉疾病的风险方面提供有价值的临床见解。应用系统生物工程方法，使用真实的时间成像和新的微流体技术，在细胞和分子尺度上研究代谢和生物力学响应，有三个具体目标：1）根据TGRL的炎症能力，量化高脂餐后个体的代谢谱。2)描述TGRL信号通路如何调节主动脉内皮的致动脉粥样硬化特征。3)使用芯片实验室分析确定与高脂血症和肥胖相关的最早期事件，这些事件增加单核细胞在致动脉粥样硬化内皮上停滞的倾向。我们的总体目标是开发测定法，该测定法精确定位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，该促炎作用将平衡从健康脉管系统转移到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.