SiglecF(HI) Marks Late-Stage Neutrophils of the Infarcted Heart: A Single-Cell Transcriptomic Analysis of Neutrophil Diversification.
SiglecF(HI) Marks Late-Stage Neutrophils of the Infarcted Heart: A Single-Cell Transcriptomic Analysis of Neutrophil Diversification.
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SiglecF（HI）标记心脏受损的晚期中性粒细胞：中性粒细胞多样化的单细胞转录组学分析。
DOI：
10.1161/jaha.120.019019
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发表时间：
2021-02-16
影响因子：
5.4
通讯作者：
King KR
中科院分区：
文献类型：
--
作者：
Calcagno DM;Zhang C;Toomu A;Huang K;Ninh VK;Miyamoto S;Aguirre AD;Fu Z;Heller Brown J;King KR
中性粒细胞被认为是心肌梗死（MI）等组织损伤的短暂第一反应者，但对其多样性或动力学知之甚少。我们永久性结扎小鼠的左冠状动脉前降支，并在MI后第1至4天或稳态时对从小鼠心脏、外周血和骨髓中分离的> 28000个中性粒细胞转录组进行单细胞RNA测序和分析。心脏中性粒细胞的无监督聚类揭示了5个主要亚群，其中3个起源于骨髓，包括表达SiglecF的迟发性粒细胞，SiglecF是一种经典用于定义嗜酸性粒细胞的标志物。SiglecFHI中性粒细胞在第1天占中性粒细胞的约25%，到MI后第4天增长至占中性粒细胞的>50%。使用荧光激活细胞分选仪分选的Ly6G+SiglecFHI和Ly6G+SiglecFLO中性粒细胞的定量聚合酶链反应进行的验证研究证实了这些群体的不同性质。为了证实细胞是嗜中性粒细胞而不是嗜酸性粒细胞，我们使加塔缺陷型小鼠（GATA加塔）梗死，并观察到相似数量的浸润性Ly6 G +SiglecFHI细胞，尽管常规嗜酸性粒细胞显著减少。与其他中性粒细胞亚群相比，Ly6G+SiglecFHI中性粒细胞表达高水平的Myc调控基因，这与寿命相关，并与MI后第4天该人群的持续性一致。总体而言，我们的数据提供了一个空间和时间图谱的中性粒细胞专业化的MI，并揭示了一个动态的促炎性心脏Ly6G+SigF+（Myc+NFκB B+）的中性粒细胞，已被忽视，因为负选择。
Neutrophils are thought to be short‐lived first responders to tissue injuries such as myocardial infarction (MI), but little is known about their diversification or dynamics. We permanently ligated the left anterior descending coronary arteries of mice and performed single‐cell RNA sequencing and analysis of >28 000 neutrophil transcriptomes isolated from the heart, peripheral blood, and bone marrow of mice on days 1 to 4 after MI or at steady‐state. Unsupervised clustering of cardiac neutrophils revealed 5 major subsets, 3 of which originated in the bone marrow, including a late‐emerging granulocyte expressing SiglecF, a marker classically used to define eosinophils. SiglecFHI neutrophils represented ≈25% of neutrophils on day 1 and grew to account for >50% of neutrophils by day 4 post‐MI. Validation studies using quantitative polymerase chain reaction of fluorescent‐activated cell sorter sorted Ly6G+SiglecFHI and Ly6G+SiglecFLO neutrophils confirmed the distinct nature of these populations. To confirm that the cells were neutrophils rather than eosinophils, we infarcted GATA‐deficient mice (∆dblGATA) and observed similar quantities of infiltrating Ly6G+SiglecFHI cells despite marked reductions of conventional eosinophils. In contrast to other neutrophil subsets, Ly6G+SiglecFHI neutrophils expressed high levels of Myc‐regulated genes, which are associated with longevity and are consistent with the persistence of this population on day 4 after MI. Overall, our data provide a spatial and temporal atlas of neutrophil specialization in response to MI and reveal a dynamic proinflammatory cardiac Ly6G+SigF+(Myc+NFϰB+) neutrophil that has been overlooked because of negative selection.