综合单细胞和空间转录组分析揭示了心肌梗死后trem2高巨噬细胞的功能和梗死边界动力学。

IF 3.7 2区 生物学 Q3 CELL BIOLOGY
Tao Xiong, Yan Chen, Chang Liu, Yaxiong Li, Yayong Zhang, Qing Chang
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引用次数: 0

摘要

本研究采用单细胞RNA测序(scRNA-seq)、空间转录组学(ST)和大量RNA测序相结合的综合方法来研究心肌梗死(MI)后复杂的细胞和分子动力学。对scRNA和ST数据进行质量控制、批量校正、降维、聚类和注释。使用Milo工具分析差异细胞丰度。使用Monocle2算法进行发育轨迹推断,并使用CellPhoneDB和NicheNet探索细胞间的通信。SCENIC分析确定了巨噬细胞亚型中的活性转录因子(TFs)。此外,反褶积用于评估细胞类型的空间分布。通过细胞通讯模式探讨不同心肌区域的功能作用。结扎左前降支(LAD)建立小鼠心肌梗死(MI)和缺血再灌注(I/R)模型。采用RT-qPCR、Western blot、免疫组织化学和免疫荧光分析分子变化。体外对AC16心肌细胞(CMs)和thp -1来源的M2巨噬细胞进行氧糖剥夺/再氧合(OGD/R)和共培养实验,研究trem2介导的作用。采用CCK-8和流式细胞术分别检测细胞活力和凋亡。该研究确定了心肌梗死后不同时间点免疫细胞类型比例的动态变化。ST在梗死区、边界区和远端区显示不同的免疫细胞浸润模式,巨噬细胞随着时间的推移逐渐浸润梗死区。功能富集分析强调了涉及不同心脏区域炎症、细胞增殖和细胞外基质重塑的关键途径。该研究还发现trem2高巨噬细胞在组织修复中起关键作用。SCENIC分析揭示了调节巨噬细胞亚型的tf,强调了它们在免疫调节和组织重建中的作用。最后,细胞间通讯分析揭示了影响免疫反应和组织修复的复杂信号网络。我们的研究结果表明,在MI和I/R模型中,IZ组Trem2的表达显著增加,并且与过表达Trem2的M2巨噬细胞共培养可显著增强OGD/R条件下AC16 CMs的增殖能力,减少凋亡,表明Trem2在I/R反应和CMs存活中起关键作用。这项全面的分析提供了mi后细胞和分子景观的详细地图,突出了免疫细胞及其调控网络的时空动态。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Integrative single-cell and spatial transcriptome analysis reveals the functions of TREM2high macrophages and infarct border dynamics post-myocardial infarction.

This study employs an integrative approach combining single-cell RNA sequencing (scRNA-seq), spatial transcriptomics (ST), and bulk RNA sequencing to investigate the complex cellular and molecular dynamics following myocardial infarction (MI). Quality control, batch correction, dimensionality reduction, clustering, and annotation were performed on scRNA and ST data. The Milo tool was used to analyze differential cell abundance. Developmental trajectory inference was conducted using the Monocle2 algorithm, and cell-cell communication was explored using CellPhoneDB and NicheNet. SCENIC analysis identified active transcription factors (TFs) in macrophage subtypes. Additionally, deconvolution was used to assess the spatial distribution of cell types. The functional roles of different myocardial regions were explored through cell communication patterns. Mouse MI and ischemia-reperfusion (I/R) models were established by ligating the left anterior descending (LAD) coronary artery. Molecular changes were analyzed using RT-qPCR, Western blot, immunohistochemistry and immunofluorescence. In vitro, AC16 cardiomyocytes (CMs) and THP-1-derived M2 macrophages were subjected to oxygen-glucose deprivation/reoxygenation (OGD/R) and co-culture experiments to study TREM2-mediated effects. Cell viability and apoptosis were assessed using CCK-8 and flow cytometry, respectively. The study identified dynamic changes in the proportions of immune cell types at different time points post-MI. ST revealed distinct immune cell infiltration patterns in the infarct, border, and remote zones, with macrophages progressively infiltrating the infarct region over time. Functional enrichment analysis highlighted key pathways involved in inflammation, cell proliferation, and extracellular matrix remodeling across different cardiac regions. The study also identified Trem2high macrophages as key players in tissue repair. SCENIC analysis uncovered TFs regulating macrophage subtypes, emphasizing their roles in immune regulation and tissue reconstruction. Finally, cell-cell communication analysis revealed complex signaling networks influencing immune responses and tissue repair. Our results demonstrated that the expressions of Trem2 were significantly increased in the IZ groups in the MI and I/R model, and co-culture with TREM2-overexpressing M2 macrophages significantly enhanced the proliferative capacity and reduced apoptosis in AC16 CMs under OGD/R conditions, indicating a critical role of Trem2 in the I/R response and CMs survival. This comprehensive analysis provides a detailed map of the cellular and molecular landscape post-MI, highlighting the temporal and spatial dynamics of immune cells and their regulatory networks.

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来源期刊
Molecular and Cellular Biochemistry
Molecular and Cellular Biochemistry 生物-细胞生物学
CiteScore
8.30
自引率
2.30%
发文量
293
审稿时长
1.7 months
期刊介绍: Molecular and Cellular Biochemistry: An International Journal for Chemical Biology in Health and Disease publishes original research papers and short communications in all areas of the biochemical sciences, emphasizing novel findings relevant to the biochemical basis of cellular function and disease processes, as well as the mechanics of action of hormones and chemical agents. Coverage includes membrane transport, receptor mechanism, immune response, secretory processes, and cytoskeletal function, as well as biochemical structure-function relationships in the cell. In addition to the reports of original research, the journal publishes state of the art reviews. Specific subjects covered by Molecular and Cellular Biochemistry include cellular metabolism, cellular pathophysiology, enzymology, ion transport, lipid biochemistry, membrane biochemistry, molecular biology, nuclear structure and function, and protein chemistry.
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