Integrated Single-Cell Atlas of Endothelial Cells of the Human Lung.
Integrated Single-Cell Atlas of Endothelial Cells of the Human Lung.
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DOI：
10.1161/circulationaha.120.052318
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发表时间：
2021-07-27
期刊：
影响因子：
37.8
通讯作者：
Kaminski N
中科院分区：
文献类型：
--
作者：
Schupp JC;Adams TS;Cosme C Jr;Raredon MSB;Yuan Y;Omote N;Poli S;Chioccioli M;Rose KA;Manning EP;Sauler M;DeIuliis G;Ahangari F;Neumark N;Habermann AC;Gutierrez AJ;Bui LT;Lafyatis R;Pierce RW;Meyer KB;Nawijn MC;Teichmann SA;Banovich NE;Kropski JA;Niklason LE;Pe'er D;Yan X;Homer RJ;Rosas IO;Kaminski N
补充数字内容可在文本中找到。人类肺内皮细胞的多样性尚未得到系统的表征。我们提供了一个人肺内皮细胞（EC）的参考图谱，以促进更好地了解肺内皮细胞的表型多样性和组成。我们重新处理了来自6个数据集的人类对照单细胞RNA测序（scRNAseq）数据。EC群体的特征在于通过迭代聚类与随后的差异表达分析。标记基因通过荧光显微镜和原位杂交进行验证。进行体外培养的原代肺EC的scRNAseq。研究了不同肺细胞类型之间的信号网络。对于跨物种分析或疾病相关性，我们将相同的方法应用于从小鼠肺或患有肺动脉高压的人肺获得的scRNAseq数据。对6个肺scRNAseq数据集进行重新分析和注释，以鉴定来自73个个体的> 15000个血管EC细胞。EC的差异表达分析揭示了对应于内皮谱系的特征，包括泛内皮、泛血管和亚群特异性标记基因集。除了淋巴管、毛细血管、动脉和静脉EC的广泛细胞类别之外，我们还发现了以前难以区分的亚群;在静脉EC中，我们发现了2个以前难以区分的群体：肺静脉EC（COL15A1阴性）定位于肺实质和体静脉EC（COL15A1pos）定位于气道和脏层胸膜;在毛细血管内皮细胞中，我们证实了它们的亚分类为最近发现的以EDNRB、SOSTDC 1和TBX 2为特征的气细胞和普通毛细血管内皮细胞。我们证实，所有6种内皮细胞类型，包括体静脉EC和气细胞，都存在于小鼠中，并确定了在人类和小鼠中保守的内皮标志物基因。配体-受体连接体分析揭示了EC与其他肺驻留细胞类型的重要稳态串扰。可商购获得的原代肺EC的scRNAseq证明在培养物中其天然肺表型的丧失。scRNAseq揭示了内皮多样性在肺动脉高压中得以维持。我们的文章附带了一个在线数据挖掘工具（www.example.com）。我们的综合分析提供了一个全面和精心制作的正常肺EC参考图谱，并确认和详细描述了大量人类和小鼠中以前未被识别的内皮细胞群。
Supplemental Digital Content is available in the text. Cellular diversity of the lung endothelium has not been systematically characterized in humans. We provide a reference atlas of human lung endothelial cells (ECs) to facilitate a better understanding of the phenotypic diversity and composition of cells comprising the lung endothelium. We reprocessed human control single-cell RNA sequencing (scRNAseq) data from 6 datasets. EC populations were characterized through iterative clustering with subsequent differential expression analysis. Marker genes were validated by fluorescent microscopy and in situ hybridization. scRNAseq of primary lung ECs cultured in vitro was performed. The signaling network between different lung cell types was studied. For cross-species analysis or disease relevance, we applied the same methods to scRNAseq data obtained from mouse lungs or from human lungs with pulmonary hypertension. Six lung scRNAseq datasets were reanalyzed and annotated to identify >15 000 vascular EC cells from 73 individuals. Differential expression analysis of EC revealed signatures corresponding to endothelial lineage, including panendothelial, panvascular, and subpopulation-specific marker gene sets. Beyond the broad cellular categories of lymphatic, capillary, arterial, and venous ECs, we found previously indistinguishable subpopulations; among venous EC, we identified 2 previously indistinguishable populations: pulmonary–venous ECs (COL15A1neg) localized to the lung parenchyma and systemic–venous ECs (COL15A1pos) localized to the airways and the visceral pleura; among capillary ECs, we confirmed their subclassification into recently discovered aerocytes characterized by EDNRB, SOSTDC1, and TBX2 and general capillary EC. We confirmed that all 6 endothelial cell types, including the systemic–venous ECs and aerocytes, are present in mice and identified endothelial marker genes conserved in humans and mice. Ligand-receptor connectome analysis revealed important homeostatic crosstalk of EC with other lung resident cell types. scRNAseq of commercially available primary lung ECs demonstrated a loss of their native lung phenotype in culture. scRNAseq revealed that endothelial diversity is maintained in pulmonary hypertension. Our article is accompanied by an online data mining tool (www.LungEndothelialCellAtlas.com). Our integrated analysis provides a comprehensive and well-crafted reference atlas of ECs in the normal lung and confirms and describes in detail previously unrecognized endothelial populations across a large number of humans and mice.