Single Cell RNA Sequencing Identifies Subsets of Hepatic Stellate Cells and Myofibroblasts in Liver Fibrosis
Single Cell RNA Sequencing Identifies Subsets of Hepatic Stellate Cells and Myofibroblasts in Liver Fibrosis
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DOI：
10.3390/cells8050503
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发表时间：
2019-05-01
期刊：
影响因子：
6
通讯作者：
Tacke, Frank
中科院分区：
文献类型：
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作者：
Krenkel, Oliver;Hundertmark, Jana;Tacke, Frank
肝星状细胞（HSC）的活化及其向胶原分泌肌成纤维细胞（MFB）的转分化促进了肝纤维化的进展。在慢性肝病期间，静息HSC被炎症和损伤信号激活。然而，HSC/MFB不仅产生胶原，还分泌细胞因子，参与代谢，并具有生物力学特性。我们在此的目的是通过单细胞RNA测序来表征这些肝脏间充质细胞的异质性。从C57 BL 6/J小鼠中分离体内静息的HSC或活化的MFB，所述C57 BL 6/J小鼠经四氯化碳（CCl 4）腹腔内激发3周以诱导肝纤维化，并与体外培养的MFB进行比较。虽然静息HSC形成了以高血小板衍生生长因子受体（PDGFR）表达为特征的同质群体，但体内和体外活化的MFB分裂成异质群体，其特征为-平滑肌肌动蛋白（-SMA）、胶原蛋白或免疫标记物。S100钙结合蛋白A6（S100 A6）是MFB激活的基因和蛋白表达水平上的通用标志物。与体内MFB的异质性相比，体外MFB在培养活化过程中顺序地且仅瞬时地表达标记基因，如趋化因子。综上所述，我们的数据表明了HSC和MFB的异质性，表明在肝纤维化中存在功能相关的亚群。
Activation of hepatic stellate cells (HSCs) and their trans-differentiation towards collagen-secreting myofibroblasts (MFB) promote liver fibrosis progression. During chronic liver disease, resting HSCs become activated by inflammatory and injury signals. However, HSCs/MFB not only produce collagen, but also secrete cytokines, participate in metabolism, and have biomechanical properties. We herein aimed to characterize the heterogeneity of these liver mesenchymal cells by single cell RNA sequencing. In vivo resting HSCs or activated MFB were isolated from C57BL6/J mice challenged by carbon tetrachloride (CCl4) intraperitoneally for 3 weeks to induce liver fibrosis and compared to in vitro cultivated MFB. While resting HSCs formed a homogenous population characterized by high platelet derived growth factor receptor (PDGFR) expression, in vivo and in vitro activated MFB split into heterogeneous populations, characterized by -smooth muscle actin (-SMA), collagens, or immunological markers. S100 calcium binding protein A6 (S100A6) was a universal marker of activated MFB on both the gene and protein expression level. Compared to the heterogeneity of in vivo MFB, MFB in vitro sequentially and only transiently expressed marker genes, such as chemokines, during culture activation. Taken together, our data demonstrate the heterogeneity of HSCs and MFB, indicating the existence of functionally relevant subsets in hepatic fibrosis.