SiglecF(HI) Marks Late‐Stage Neutrophils of the Infarcted Heart: A Single‐Cell Transcriptomic Analysis of Neutrophil Diversification

Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Aaron D. Aguirre¹  Claire Zhang²  Avinash Toomu²  David M. Calcagno²  Kevin R. King²  Shigeki Miyamoto³  Joan Heller Brown³  Van K. Ninh³  Kenneth Huang⁴  Zhenxing Fu⁴ 

[1] Cardiology Division Center for Systems Biology Wellman Center for Photomedicine Massachusetts General Hospital Boston MA;Department of Bioengineering Jacobs School of Engineering University of California San Diego La Jolla CA;Department of Pharmacology University of California San Diego La Jolla CA;Division of Cardiology and Cardiovascular Institute Department of Medicine University of California San Diego La Jolla CA;

关键词: granulopoiesis;    heart;    myocardial infarction;    neutrophil maturity;    neutrophils;    siglecF;   

DOI  :  10.1161/JAHA.120.019019

来源: DOAJ

【 摘 要 】

Background 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. Methods and Results 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. Conclusions 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.

【 授权许可】

Unknown