Integrated Analysis of Hematopoietic Precursor Self-renewal and Differentiation

该提案的目标是激活独立阶段（ROO），与 原裁决书，即：to generate生成a novel新and Integrated综合view视图of mechanisms机制of 使用EML（红细胞、髓细胞和淋巴细胞）造血前体自我更新和分化 多能细胞作为模型系统。EML细胞是研究早期免疫缺陷的分子控制的理想细胞。 大规模造血分化。EML细胞产生自我更新的CD 34+前体细胞 和部分分化的非更新CD 34-细胞。可以生长大量的EML细胞， 在缺乏解剖学生态位的情况下在体外分化。基于我的K99阶段差异化研究 基因表达和转录因子结合使用芯片测序，我假设有关键 转录调节网络中的调节因子决定EML细胞自我更新和 分化，如TCF 7和RUNX 1。我已经构建了初步的转录调控 电路由TCF 7和RUNXI调节。对于我的ROO研究阶段，我计划在全球范围内确定关键 利用基因表达调控EML细胞自我更新和分化的转录调节因子， 蛋白质组学数据来指导转录调控工作。结合靶和转录因子将 我将确定目标枢纽，并测试主要监管机构。 随后，我将把我们的基因组学、蛋白质组学、磷酸化数据和文献整合到 转录因子结合网络，并进一步发展全球相互作用网络。最后我会确认 人类原代细胞的关键发现。拟议的项目可以导致分子和生物化学研究， 我自己的实验室

The goal of this proposal for activafing the independent phase (ROO) stays the same as that of the original proposal for the award, i.e., to generate a novel and Integrated view ofthe mechanisms of hematopoietic precursor self-renewal and differentiation using EML (erythroid, myeloid, and lymphocytic) multipotenfial cells as a model system. EML cells are ideal for studying the molecular control of eariy hematopoietic differentiation at a large scale. EML cells give rise to the self-renewing CD34+ precursor cells and partially differenfiated non-renewing CD34- cells. Large quantities of EML cells can be grown and differentiated in vitro in the absence of an anatomical niche. Based on my K99 phase of study in differenfial gene expression, and of transcription factor binding using Chip-Sequencing, I hypothesize that there are key regulators in transcriptional regulatory networks determining the choice between EML cell self-renewal and differentiation, such as TCF7 and RUNX1. I have already constmcted preliminary transcripfional regulatory circuits regulated by TCF7 and RUNXI. For my ROO phase of research, 1 plan to globally identify the key transcriptional regulators controlling EML cell self-renewal and differenfiafion by using gene expression and proteomic data to guide the transcripfional regulafion work. The binding targets and transcription factors will be assembled into regulatory networks and I will identify target hubs and test for master regulators. Subsequenfiy I will integrate our genomic, proteomics, phosphorylafion data and literature into the transcripfion factor binding networks and further develop a global interaction network. Finally I will confirm key findings in human primary cells. The proposed project can lead to molecular and biochemical studies in my own lab for many years to come.