The Hematopoietic Kinome

描述(申请人提供)：在过去的两年里，阵列和质谱学技术已经能够分析造血干细胞(HSC)及其下游祖细胞的转录组和蛋白质组。这些信息将对阐明控制干细胞生理学和血细胞分化的分子机制具有重要价值。然而，一个同样重要的目标，如果不是更重要的话，是定义那些参与在造血干细胞及其后代中活跃的信号通路的蛋白质。其他蛋白质上酪氨酸、丝氨酸和苏氨酸残基的磷酸化酶在决定细胞周期进入、存活和哺乳动物体内细胞分化命运的信号级联中发挥着重要作用，包括造血系统。特别是，了解在HSC及其分化后代中活跃的信号通路将为理解干细胞生物学和细胞分化提供关键信息。传统的遗传和生化方法当然可以提供其中的一些答案；然而，出于技术和实践的原因，这些方法通常一次只能追求一个基因或途径。因此，需要一种更全面的方法来揭示活跃在HSC及其分化后代中的信号通路。为此，我们开发了基因组分析技术，并开始将它们应用于HSC和分化的祖细胞类型。在这项研究中，我们建议提供一个全面的分析，因为这将进一步定义我们对造血发育不同阶段所使用的信号通路的理解。这项建议将在以下具体目标中进行：目标1：比较和对比稳态造血过程中LT-HSC和ST-HSC群体的基因组。目的2：测定造血室扩增过程中的LT-HSC动态组。目的3：确定关键的多能祖细胞和谱系受限的祖细胞类型的基因组。

DESCRIPTION (provided by applicant): Over the last 2 years array and mass spectrometry technologies have enabled analysis of the transcriptome and proteome of hematopoietic stem cells (HSC) and their downstream progenitors. This information will be of significant value to the elucidation of molecular mechanisms that govern stem cell physiology and blood cell differentiation. However, an equally, if not more important goal, is to define those proteins that participate in signaling pathways that are active in hematopoietic stem cells and their progeny. Enzymes that phosphorylate tyrosine, serine and threonine residues on other proteins play a major role in signaling cascades that determine cell cycle entry, survival and the differentiation fate of cells in the mammalian body, including the hematopoietic system. In particular, knowing the signaling pathways that are active in HSC and their differentiated progeny will provide critical information for understanding stem cell biology and cell differentiation. Traditional genetic and biochemical approaches can certainly provide some of these answers; however, for technical and practical reasons, these are typically pursued one gene or pathway at a time. Thus, a more comprehensive approach is needed in order to reveal signaling pathways active in HSC and their differentiated progeny. Towards this end, we have developed kinome analysis techniques and begun to apply them to HSC and differentiated progenitor cell types. In this study, we propose to provide a comprehensive analysis of the HSC kinome and their more differentiated progeny as this will further define our understanding of the signaling pathways utilized during different stages of hematopoietic development. This proposal will be pursued in the following specific aims: Aim 1: Compare and contrast the kinome of LT-HSC and ST-HSC populations during steady-state hematopoiesis. Aim 2: Determine the LT-HSC kinome during expansion of the hematopoietic compartment. Aim 3: Determine the kinome of key multi-potent progenitors and lineage-restricted progenitor cell types.