Cytokines and lineage choice in hematopoietic precursors

描述（由申请人提供）：人CD34+造血前体细胞可以很容易地从成人外周血中获得。用各种细胞因子混合处理这些细胞可以诱导红细胞、巨核细胞或髓系分化的细胞高效产生。我们发现不同的细胞因子鸡尾酒诱导快速和广泛的基因表达变化模式，不同的细胞因子鸡尾酒不同。这些显著的变化表明，它们发生在CD34+细胞的很大一部分，并可能调节单个细胞的分化模式。最近发展的方法可以全面测量单细胞中的mRNA表达和DNA甲基化，这使得在特异性水平上研究细胞因子在谱系促进中的作用成为可能。在目前的应用中，我们建议使用单细胞基因组规模分析和细胞标记和标记方法的组合来确定相同的细胞是否可以对不同的细胞因子处理产生不同的反应，并研究这些不同的反应是否会导致不同谱系的分化。最终目标是确定谱系特异性细胞因子的早期转录反应的性质，以及这些反应是否具有指导性和允许谱系规范。

DESCRIPTION (provided by applicant): Human CD34+ hematopoietic precursor cells can readily be obtained from the peripheral blood of adults. Treatment of these cells with various cytokine cocktails can induce efficient production of cells differentiated along erythroid, megakaryocytic or myeloid lines. We find that different cytokine cocktails induce rapid and extensive patterns of change in gene expression that are different for different cytokine cocktails. The prominence of these changes suggests that they occur in a large fraction of the CD34+ cells and may modulate the patterns of differentiation of individual cells. The recent development of methods that permit global measurements of mRNA expression, and DNA methylation in single cells make it possible to investigate the role of cytokines in lineage promotion at a level of specificity hitherto unavailable. In the present application we propose to use a combination of single cell genomic scale analyses, and cell tagging and labeling approaches to establish if the same cell can respond differently to different cytokine treatments, and to investigate whether these different responses result in differentiation along alternate lineages. The ultimate goal is to determine the nature of early transcriptional responses to lineage specific cytokines and whether these are instructional as well as permissive for lineage specification.