DIFFERENTIATION OF GRANULOCYTES AND MACROPHAGES

这个项目的长期目标是了解介导细胞增殖和分化的细胞因子信号通路的生理控制。 这些途径被细胞因子不适当地激活，或者在包括癌症、自身免疫以及炎性和感染性疾病在内的广泛疾病中被破坏。 在此资助下，以前的工作使用功能性遗传筛选确定了一个细胞因子信号传导的关键负调节因子家族，称为细胞因子信号传导抑制因子（SOCS）。 基因缺失实验表明，SOCS-1是一个必要的分子，需要保持在检查内源性干扰素-γ和生化实验的潜在致死作用揭示，SOCS-1与激活的JAK激酶相互作用，并抑制其活性。 本项目的目的是：（1）在基因缺失小鼠中分析三种不同的SOCS蛋白（SOCS-1至-3）的独特和冗余的生理作用。 在致死表型的情况下（如SOCS-1 -/-和SOCS-3-/-小鼠），将进行组织特异性、条件性基因缺失或与精氨酸缺失小鼠杂交，以鉴定缺陷的作用位点。(2)通过对具有不同SOCS或细胞因子缺失的小鼠进行遗传杂交来确定不同SOCS蛋白在抑制不同细胞因子的作用中的特异性;以及（3）鉴定与SOCS蛋白相互作用以介导其抑制作用的生理分子靶标，并确定每种相互作用的生物化学和生物学后果。 将通过共缔合、直接结合和肽特异性识别的定义来鉴定相互作用蛋白，并确定每种相互作用的定量结合亲和力。所有基于体外生物学效应或生化相互作用的分析将通过产生适当的基因缺失或修饰小鼠（例如，产生仅缺失一个相互作用结构域的SOCS蛋白）和通过适当的遗传杂交（例如，与缺失拟定相互作用伴侣的小鼠）在整个动物中进行验证。 预期这些研究将定义生理学验证的治疗靶标，用于设计在治疗与细胞因子信号传导途径的外源性或内源性活化相关的疾病（特别是炎症、白血病和其他癌症）中应具有治疗价值的分子。

The long term aim of this project is to understand the physiological control of cytokine signaling pathways that mediate cellular proliferation and differentiation. These pathways are activated inappropriately by cytokines or are otherwise subverted in a wide range of diseases including cancer, autoimmunity and inflammatory and infectious diseases. Previous work funded under this grant using functional genetic screens identified a family of critical negative regulators of cytokine signaling called the supressors of cytokine signaling (SOCS). Gene deletion experiments revealed that SOCS-1 is an essential molecule required to keep in check the potentially lethal effects of endogenous interferon-gamma and biochemical experiments revealed that SOCS-1 interacts with activated JAK kinases and inhibits their activity. The aims of the present project are: (1) to analyse in gene-deleted mice the unique and redundant physiological roles of three different SOCS proteins (SOCS-1 to - 3). In the case of a lethal phenotype (as for SOCS-1 -/- and SOCS-3-/- mice) tissue-specific, conditional gene deletions or crosses to cytokine-deleted mice will be performed to identify the site of action of the defect. (2) to determine the specificity of different SOCS proteins in inhibiting the actions of different cytokines by performing genetic crosses of mice with different SOCS or cytokine deletions; and (3) to identify the physiological molecular targets that interact with SOCS proteins to mediate their inhibitory effects and determine the biochemical and biological consequences of each interaction. Interacting proteins will be identified by co-association, direct binding and definition of peptide specificity recognition and the quantitative binding affinity of each interaction determined. All analyses based on in vitro biological effects or biochemical interactions will be verified in the whole animal by generating appropriate gene-deleted or -modified mice (e.g. producing a SOCS protein with only one interaction domain deleted) and by appropriate genetic crosses (e.g with mice in which the proposed interaction partner has been deleted). These studies are expected to define physiologically-validated therapeutic targets for the design of molecules that should have therapeutic value in treating diseases associated with the exogenous or endogenous activation of cytokine signaling pathways, especially inflammation, leukemias and other cancers.