Cytokines and lineage choice in hematopoietic precursors

摘要：人CD34造血祖细胞可以很容易地从人外周血中获得。 成年人。用不同的细胞因子鸡尾酒处理这些细胞可以诱导高效地产生细胞 沿着红系、巨核系或髓系分化。我们发现不同的细胞因子鸡尾酒诱导 基因表达的快速和广泛的变化模式对于不同的细胞因子鸡尾酒是不同的。 这些变化的突出表明，它们发生在CD34细胞的很大一部分中，并可能 调节单个细胞的分化模式。方法的最新发展允许 对单个细胞中mRNA表达和DNA甲基化的全球测量使研究成为可能 细胞因子在谱系促进中的作用达到了迄今尚无的特异性水平。在本申请中 我们建议结合使用单细胞基因组规模分析、细胞标记和标记 确定同一细胞是否可以对不同的细胞因子治疗有不同反应的方法，以及 研究这些不同的反应是否会导致不同血统的分化。终极的 目的是确定对谱系特异性细胞因子的早期转录反应的性质，以及 这些是指导性的，也是世系规范的许可。

Abstract: 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.