DataSheet_1_Single-Cell RNA Sequencing of Peripheral Blood Mononuclear Cells From Acute Myocardial Infarction.doc

BackgroundAcute myocardial infarction (AMI) can occur in patients with atherosclerotic disease, with or without plaque rupture. Previous studies have indicated a set of immune responses to plaque rupture. However, the specific circulating immune cell subsets that mediate inflammatory plaque rupture remain elusive.
MethodsTen AMI patients were enrolled in our study (five with and five without plaque rupture; plaque characteristics were identified by optical coherence tomography). By single-cell RNA sequencing, we analyzed the transcriptomic profile of peripheral blood mononuclear cells.
ResultsWe identified 27 cell clusters among 82,550 cells, including monocytes, T cells, NK cells, B cells, megakaryocytes, and CD34+ cells. Classical and non-classical monocytes constitute the major inflammatory cell types, and pro-inflammatory genes such as CCL5, TLR7, and CX3CR1 were significantly upregulated in patients with plaque rupture, while the neutrophil activation and degranulation genes FPR2, MMP9, and CLEC4D were significantly expressed in the intermediate monocytes derived from patients without plaque rupture. We also found that CD4+ effector T cells may contribute to plaque rupture by producing a range of cytokines and inflammatory-related chemokines, while CD8+ effector T cells express more effector molecules in patients without plaque rupture, such as GZMB, GNLY, and PRF1, which may contribute to the progress of plaque erosion. Additionally, NK and B cells played a significant role in activating inflammatory cells and promoting chemokine production in the plaque rupture. Cell–cell communication elaborated characteristics in signaling pathways dominated by inflammatory activation of classical monocytes in patients with plaque rupture.
ConclusionsOur studies demonstrate that the circulating immune cells of patients with plaque rupture exhibit highly pro-inflammatory characteristics, while plaque erosion is mainly associated with intermediate monocyte amplification, neutrophil activation, and degranulation. These findings may provide novel targets for the precise treatment of patients with AMI.

背景 急性心肌梗死（acute myocardial infarction, AMI）可发生于伴或不伴斑块破裂的动脉粥样硬化性疾病患者中。既往研究已揭示斑块破裂存在一系列免疫应答过程，但介导炎症性斑块破裂的特异性循环免疫细胞亚群仍尚不明确。
方法 本研究纳入10例AMI患者（其中伴斑块破裂与不伴斑块破裂者各5例；斑块特征通过光学相干断层扫描（optical coherence tomography）明确）。采用单细胞RNA测序技术分析外周血单个核细胞的转录组特征。
结果 本研究在82550个细胞中鉴定出27个细胞群，涵盖单核细胞、T细胞、自然杀伤（natural killer, NK）细胞、B细胞、巨核细胞及CD34+细胞。经典型与非经典型单核细胞为主要的炎症细胞类型；伴斑块破裂患者中，CCL5、TLR7及CX3CR1等促炎基因显著上调，而不伴斑块破裂患者来源的中间型单核细胞中，中性粒细胞活化与脱颗粒相关基因FPR2、MMP9及CLEC4D呈显著高表达。本研究同时发现，CD4+效应T细胞可通过分泌多种细胞因子及炎症相关趋化因子参与斑块破裂过程；而不伴斑块破裂患者中，CD8+效应T细胞表达更多效应分子，如GZMB、GNLY及PRF1，此类分子可能参与斑块糜烂的进展。此外，在斑块破裂过程中，NK细胞与B细胞在活化炎症细胞及促进趋化因子产生方面发挥重要作用。细胞间互作分析显示，伴斑块破裂患者的信号通路以经典型单核细胞的炎症活化为主导，呈现特征性改变。
结论 本研究证实，伴斑块破裂患者的循环免疫细胞呈现显著的促炎特征，而斑块糜烂主要与中间型单核细胞扩增、中性粒细胞活化及脱颗粒相关。本研究结果可为AMI患者的精准治疗提供全新靶点。