Integrated Analysis of Hematopoietic Precursor Self-renewal and Differentiation

简介(申请人提供)：我是耶鲁大学迈克尔·斯奈德博士实验室的Ruth L.Kirschstein国家研究服务奖博士后研究员。我之前的培训重点是分子遗传学和基因组学，特别是全基因组转录图谱和新的基因特征。在拟议的职业发展期间，我将在实验和计算层面上接受造血、功能基因组学、蛋白质组学、大规模数据集成和网络建设方面的深入培训。耶鲁大学基因组和蛋白质组学中心良好的培训环境，再加上我的共同导师Sherman Weissman博士和咨询委员会在造血干细胞生物学方面的专业知识，非常适合我在细胞自我更新和分化方面开始独立的学术研究生涯。我的五年研究计划的目标是通过使用整合的基因组和蛋白质组方法，以小鼠EML(红系、髓系和淋巴系)多潜能造血祖细胞作为主要模型，产生一个新的和全面的细胞自我更新和分化的观点。EML细胞是大规模研究早期造血分化的分子调控的理想细胞。EML细胞产生自我更新的CD34+前体细胞和部分分化的非更新的CD34-细胞。大量的EML细胞可以在没有解剖学定位的情况下在体外培养和分化。基于我对差异基因表达谱和利用芯片结合转录因子的初步研究，我假设在转录调控网络中存在关键的调控因子(如：Tcf7、RUNX1和GATA2)，它们决定了EML细胞自我更新和分化之间的选择。具体地说，我建议通过使用基因表达和蛋白质组数据来指导转录调控工作，从而在全球范围内识别控制这些过程的关键转录调控因子。结合靶标和转录因子将被组装成调控网络，我将确定靶标枢纽并测试主要调节者。随后，我将把我们的基因组、蛋白质组、磷酸化数据和文献整合到转录因子结合网络中，并进一步发展一个全球相互作用网络。最后，我将确认在人类造血祖细胞中的关键发现。相关性(参见说明)：这些对EML细胞的研究将展示干细胞可利用的自我更新和分化机制的基本特性，这些机制在修复或再生受损的组织和器官方面具有巨大的前景。通过这项研究获得的分子理解有望提高我们指导造血干细胞命运的能力，例如，通过在体外复制和分化HSCs。因此，本研究对白血病的公共卫生和治疗具有重要的参考价值。

DESCRIPTION (provided by applicant): I am a Ruth L. Kirschstein National Research Service Award postdoctoral fellow in Dr. Michael Snyder's lab at Yale University. My previous training focused on molecular genetics and genomics, specifically genome-wide transcriptional mapping and novel gene characterization. During the proposed career development I will obtain in-depth training in hematopoiesis, functional genomics, proteomics, large-scale data integration and network construction on both experimental and computational levels. The excellent training environment in the Yale Center for Genomics and Proteomics in combination with my co-mentor Dr. Sherman Weissman and advisory committee's expertise in hematopoietic stem cell biology is uniquely suited for me to launch an independent academic research career in cell self-renewal and differentiation. The goal of my five-year research plan is to generate a novel and comprehensive view of cell self-renewal and differentiation by using integrated genomic and proteomic approaches and murine EML (Erythroid, Myeloid, and Lymphocytic) multipotential hematopoietic precursor cells as a primary model. EML cells are ideal for studying the molecular control of early hematopoietic differentiation at a large scale. EML cells give rise to the self-renewing CD34+ precursor cells and partially differentiated 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 preliminary study of differential gene expression profiling, and of transcription factor binding using Chip-chip, I hypothesize that there are key regulators (such as: TCF7, RUNX1 and GATA2) in transcriptional regulatory networks that determine the choice between EML cell self-renewal and differentiation. Specifically, I propose to globally identify the key transcriptional regulators controlling these processes by using gene expression and proteomic data to guide the transcriptional regulation work. The binding targets and transcription factors will be assembled into regulatory networks, and I will identify target hubs and test for master regulators. Subsequently I will integrate our genomic, proteomic, phosphorylation data and literature into the transcription factor binding networks and further develop a global interaction network. Finally I will confirm key findings in human hematopoietic precursor cells. RELEVANCE (See instructions): These studies in EML cells will demonstrate fundamental properties of self-renewal and differentiation mechanisms available to stem cells which hold great promise in repairing or regenerating damaged tissues and organs. Molecular understanding gained through this study will hopefully improve our ability to direct hematopoietic stem cell fate by, for example, replicating and differentiating HSCs in vitro. Therefore, this study is highly valuable for public health and therapeutic purposes for leukemia.