Single-cell transcriptional analysis of normal, aberrant, and malignant hematopoiesis in zebrafish.
Single-cell transcriptional analysis of normal, aberrant, and malignant hematopoiesis in zebrafish.
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DOI：
10.1084/jem.20152013
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发表时间：
2016-05-30
期刊：
影响因子：
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通讯作者：
Langenau DM
中科院分区：
文献类型：
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作者：
Moore FE;Garcia EG;Lobbardi R;Jain E;Tang Q;Moore JC;Cortes M;Molodtsov A;Kasheta M;Luo CC;Garcia AJ;Mylvaganam R;Yoder JA;Blackburn JS;Sadreyev RI;Ceol CJ;North TE;Langenau DM
Moore等人报道了斑马鱼血液中首次单细胞基因表达分析，以区分主要血统，鉴定新的细胞类型，并描述T细胞白血病的异质性。造血在产生功能不同的血细胞类型时达到高潮。在斑马鱼中，缺乏细胞表面抗体迫使研究人员使用荧光转基因报告系来标记特定的血细胞部分。然而，这些方法受到可区分的转基因系和荧光蛋白组合的可用性的限制。在这里，我们对来自斑马鱼的单个造血细胞进行转录分析，以确定红系、髓系、B细胞和T细胞谱系。我们还使用我们的方法鉴定了造血干细胞和祖细胞以及一种新的NK-裂解素4+细胞类型，代表了假定的细胞毒性T/NK细胞。我们的平台还量化了rag2E450fs突变鱼的造血缺陷，并显示这些鱼的T细胞减少，随后NK-lysin 4+细胞和髓细胞增加。这些数据提示了rag2E450fs突变斑马鱼先天免疫系统的代偿调节。最后，对myc诱导的T细胞急性淋巴母细胞白血病的分析表明，细胞停留在CD4+/CD8+皮质胸腺细胞阶段，并且一部分白血病细胞不恰当地重新表达干细胞基因，包括bmi1和cmyb。总的来说，我们的实验为斑马鱼血液和白血病中的单细胞异质性提供了新的工具和生物学见解。
Moore et al. reports the first single-cell gene expression analysis in zebrafish blood to distinguish major blood lineages, identify new cell types, and delineate heterogeneity in T cell leukemia. Hematopoiesis culminates in the production of functionally heterogeneous blood cell types. In zebrafish, the lack of cell surface antibodies has compelled researchers to use fluorescent transgenic reporter lines to label specific blood cell fractions. However, these approaches are limited by the availability of transgenic lines and fluorescent protein combinations that can be distinguished. Here, we have transcriptionally profiled single hematopoietic cells from zebrafish to define erythroid, myeloid, B, and T cell lineages. We also used our approach to identify hematopoietic stem and progenitor cells and a novel NK-lysin 4+ cell type, representing a putative cytotoxic T/NK cell. Our platform also quantified hematopoietic defects in rag2E450fs mutant fish and showed that these fish have reduced T cells with a subsequent expansion of NK-lysin 4+ cells and myeloid cells. These data suggest compensatory regulation of the innate immune system in rag2E450fs mutant zebrafish. Finally, analysis of Myc-induced T cell acute lymphoblastic leukemia showed that cells are arrested at the CD4+/CD8+ cortical thymocyte stage and that a subset of leukemia cells inappropriately reexpress stem cell genes, including bmi1 and cmyb. In total, our experiments provide new tools and biological insights into single-cell heterogeneity found in zebrafish blood and leukemia.