Dissecting the cellular and molecular immune interactions in human atherosclerosis

项目摘要 由于动脉粥样硬化引起的心血管疾病（CVD）是死亡的主要原因， 全球残疾人。即使在接受优化标准治疗方案治疗的患者中， 发病率和死亡率仍然很高。直接针对动脉粥样硬化的新策略- 心血管疾病的主要原因-是迫切需要的。虽然最近的概念进展 强调了慢性炎症和免疫系统激活的重要性， 心血管疾病中的动脉粥样硬化，在随机临床试验中测试的新的有前途的抗炎药 试验显示，在降低心血管终点或动脉粥样硬化斑块方面没有益处 负担免疫系统由一组分层组织的分子和 在组织和系统中协同作用的细胞网络。然而，大多数关于 人类动脉粥样硬化中免疫系统集中在循环细胞上-很少或没有 重点关注组织部位的细胞免疫浸润或它们之间的关系。获得 更好地了解有助于CVD的全身和局部免疫网络，我们将 利用分子系统生物学的技术进步。使用CyTOF质量 流式细胞术-一种用于免疫细胞高维分析的尖端技术-我们 循环外周血的确定的单细胞变异和分子活化 单核细胞（PBMC）和动脉粥样硬化组织相关免疫细胞在患者 接受外科血管重建术的颈动脉疾病。我们将使用多组织系统 生物学方法来推断动脉壁和血液特异性免疫网络， 免疫反应在动脉粥样硬化的不同阶段。我们将联合收割机的使用创新 CyTOF和RNA下一代（NGS）等技术，以识别免疫调节 与人类CVD疾病相关的网络。在具体目标1中，我们将识别单细胞 循环和组织相关的人免疫细胞的组成和分子状态。我们 还将确定系统和组织免疫之间的分子和细胞相关性， 细胞多样性和动脉粥样硬化和临床表型。在具体目标2中，我们将确定 动脉粥样硬化中的免疫网络和已鉴定免疫网络的临床相关性 在动脉粥样硬化中被激活。利用系统生物学研究， 测量患者样本中免疫细胞变异以推断免疫调节的技术 网络将加深我们对免疫系统对CVD的贡献的认识， 为新型抗炎疗法的分子靶点选择奠定了基础。

PROJECT SUMMARY Cardiovascular disease (CVD) due to atherosclerosis is the leading cause of mortality and disability globally. Even in patients treated with optimized standard-of-care regimens, residual morbidity and mortality remain high. New strategies that directly target atherosclerosis—the main cause of CVD—are urgently needed. Although recent conceptual advances have emphasized the importance of chronic inflammation and immune system activation in atherosclerosis in CVD, new promising anti-inflammatory agents tested in randomized clinical trials showed no benefits in either reducing cardiovascular end points or atherosclerotic plaque burden. The immune system is composed of a hierarchically organized set of molecular and cellular networks that act in concert in tissues and systemically. Yet, most studies of the role of immune system in atherosclerosis in humans have focused on circulating cells—with little or no focus on cellular immune infiltrates at the tissue site or the relationship between them. To gain a better understanding of systemic and local immune networks that contribute to CVD, we will take advantage of technological advances in molecular systems biology. Using CyTOF mass cytometry—a cutting-edge technique for high-dimensional analysis of immune cells—we identified single-cell variation and molecular activation of circulating peripheral blood mononuclear cells (PBMCs) and atherosclerotic tissue-associated immune cells in patients with carotid artery disease undergoing surgical revascularization. We will use a multi-tissue, systems biology approach to infer arterial wall- and blood-specific immune networks that govern the immune response at different stages of atherosclerosis. We will combine the use of innovative technologies such as CyTOF and RNA next-generation (NGS) to identify immune regulatory network relevant to human CVD disease. In specific Aim 1, we will identify the single-cell composition and molecular state of circulating and tissue-associated human immune cells. We will also determine the molecular and cellular correlates between systemic and tissue immune cell diversity and atherosclerotic and clinical phenotypes. In Specific Aim 2, we will identify immune networks in atherosclerosis and the clinical correlates of identified immune networks activated in atherosclerosis. The use of systems biology studies that integrate cutting-edge technologies to measure immune cell variation in patient samples to infer immune regulatory networks will deepen our knowledge of the contribution of the immune system to CVD and will set the stage to the selection of molecular targets for novel anti-inflammatory therapies.