Atlas of the Immune Cell Repertoire in Mouse Atherosclerosis Defined by Single-Cell RNA-Sequencing and Mass Cytometry.
Atlas of the Immune Cell Repertoire in Mouse Atherosclerosis Defined by Single-Cell RNA-Sequencing and Mass Cytometry.
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DOI：
10.1161/circresaha.117.312513
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发表时间：
2018-06-08
影响因子：
20.1
通讯作者：
Wolf D
中科院分区：
文献类型：
--
作者：
Winkels H;Ehinger E;Vassallo M;Buscher K;Dinh HQ;Kobiyama K;Hamers AAJ;Cochain C;Vafadarnejad E;Saliba AE;Zernecke A;Pramod AB;Ghosh AK;Anto Michel N;Hoppe N;Hilgendorf I;Zirlik A;Hedrick CC;Ley K;Wolf D
动脉粥样硬化是一种慢性炎症性疾病，由主动脉中促炎和抗炎白细胞的相互作用驱动。然而，主动脉白细胞的表型和转录多样性知之甚少。我们通过单细胞RNA测序（scRNAseq）和质谱仪（CyTOF）深入表征了健康和动脉粥样硬化小鼠动脉瘤的白细胞，以定义动脉粥样硬化中免疫细胞景观的图谱。使用来自喂食Apoe−/−和Ldlr−/−小鼠的食物（CD）和西方饮食（WD）的主动脉白细胞的scRNAseq，我们检测到11个具有不同表型和空间特征的主要白细胞簇，而健康大鼠的细胞库则不那么多样化。在单细胞水平上的基因集富集分析确定了多个途径，如脂质代谢、增殖和细胞因子分泌，仅限于特定的白细胞簇。动脉粥样硬化Apoe−/−和Ldlr−/−小鼠的白细胞群体受到不同的调节。我们证实了这些集群的表型多样性与一种新的CyTOF 35标记面板与金属标记的抗体和传统的流式细胞术。通过这些基于蛋白质的方法检索的细胞群与转录定义的簇高度相关。在scRNAseq、CyTOF和FACS的综合筛选策略中，我们检测到三种主要的B细胞亚群在表面标志物、功能途径和体外细胞因子分泌方面发生了变化。最后，我们使用白细胞簇基因签名，通过遗传去卷积策略来计算126个人类斑块中的白细胞频率。这种方法揭示了人颈动脉斑块和显微解剖的小鼠斑块主要由巨噬细胞、T细胞和单核细胞构成。此外，颈动脉斑块中遗传定义的白细胞群体的频率预测患者的心血管事件。通过高维分析对白细胞多样性的定义使主动脉白细胞亚群的细粒度分析成为可能，揭示了新的免疫机制和细胞类型特异性通路，并建立了人类动脉粥样硬化病变白细胞的功能相关性。
Atherosclerosis is a chronic inflammatory disease that is driven by the interplay of pro- and anti-inflammatory leukocytes in the aorta. Yet, the phenotypic and transcriptional diversity of aortic leukocytes is only poorly understood. We characterized leukocytes from healthy and atherosclerotic mouse aortas in-depth by single cell RNA-sequencing (scRNAseq) and mass cytometry (CyTOF) to define an atlas of the immune cell landscape in atherosclerosis. Employing scRNAseq of aortic leukocytes from chow (CD) and western diet (WD) fed Apoe−/− and Ldlr−/− mice, we detected 11 principal leukocyte clusters with distinct phenotypical and spatial characteristics, while the cellular repertoire in healthy aortas was less diverse. Gene set enrichment analysis on a single cell level established that multiple pathways, such as for lipid metabolism, proliferation, and cytokine secretion, were confined to particular leukocyte clusters. Leukocyte populations were differentially regulated in atherosclerotic Apoe−/− and Ldlr−/− mice. We confirmed the phenotypic diversity of these clusters with a novel CyTOF 35-marker panel with metal-labelled antibodies and conventional flow cytometry. Cell populations retrieved by these protein-based approaches were highly correlated to transcriptionally defined clusters. In an integrated screening strategy of scRNAseq, CyTOF, and FACS, we detected three principal B-cell subsets with alterations in surface markers, functional pathways, and in vitro cytokine secretion. Finally, we used leukocyte cluster gene signatures to enumerate leukocyte frequencies in 126 human plaques by a genetic deconvolution strategy. This approach revealed that human carotid plaques and microdissected mouse plaques were mostly populated by macrophages, T-cells, and monocytes. In addition, the frequency of genetically defined leukocyte populations in carotid plaques predicted cardiovascular events in patients. The definition of leukocyte diversity by high-dimensional analyses enables a fine-grained analysis of aortic leukocyte subsets, reveals new immunological mechanisms and cell-type specific pathways, and establishes a functional relevance for lesional leukocytes in human atherosclerosis.