ROLE OF FPS/FES PROTO-ONCOGENE IN MYELOID CELL FUNCTION

FPS/FES 原癌基因在骨髓细胞功能中的作用

• 批准号: 6375911
• 负责人: RICARDO A FELDMAN
• 金额: $ 25.91万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-07-01 至 2003-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6375911
• 关键词: active sites; antireceptor antibody; biological signal transduction; cell adhesion; cell growth regulation; cell line; crosslink; cytokine; enzyme activity; enzyme mechanism; genetically modified animals; immunofluorescence technique; integrins; laboratory mouse; leukocyte activation /transformation; macrophage; molecular cloning; myeloid stem cell; phosphorylation; protein tyrosine kinase; protooncogene; transcription factor; transfection

The long-term objective of this study is to understand the biological role and mechanism of action of the c-fps/Fes proto-oncogene. Fes encodes a cytoplasmic tyrosine kinase that is specifically expressed in myeloid cells. We have obtained evidence that Fes is involved in signaling through a specialized class of surface receptors during cell- cell and cell-matrix interactions, and suggests a possible role of Fes in mediating the release of inflammatory cytokines during macrophage activation. We found that VLA-4 integrin engagement in macrophages, a process that induces the release of inflammatory mediators, results in the activation of Fes kinase. We have also identified a number of macrophage specific substrates of Fes, which are involved in cell adhesion and regulation of inflammatory cytokines. One of the substrates phosphorylated by Fes is the crk-associated substrates (Cas) and another is a new 130 kDa protein (P130). Cas is an adapter protein involved in integrin signalling. P130 is a protein expressed only in hematopoietic cells and has been implicated in release of inflammatory cytokines. We hypothesize that Fes is involved in a novel integrin- dependent pathway that regulates the release of inflammatory mediators. In this proposal we will study the mechanism by which integrin engagement results in activation of Fes kinase, and how phosphorylation of its specific substrates regulates the inflammatory response of macrophages. The macrophage adhesion receptor VLA-4 will be engaged by cross-linking with specific antibodies and by adhesions to fibronectin, and the functional interactions between Fes and the integrin cascade will be examined. Using a combination of genetic and biochemical approaches we will elucidate the mechanisms involved in Fes kinase activation, characterize the Fes substrates, identify its downstream targets, and analyze the role of Fes in the adhesion-dependent macrophage inflammatory response. Finally, we will identify the transcriptional regulators activated by Fes and map the Fes-responsive elements in the promoters of genes under Fes control. Macrophages are a central player in the immune response to microorganisms, in tumoricidal activity, and in pathological conditions such as atherosclerosis. The proposed investigation will bring to light a previously unknown regulatory cascade involved in inflammatory response of macrophage. This will have an impact on the rational design of therapies aimed at preventing the inappropriate activation of macrophages, a biomedical problem of paramount importance.

这项研究的长期目标是了解 C-fps/Fes原癌基因的作用和作用机制。FeS 编码一种胞质酪氨酸激酶，它在 髓系细胞。我们已经得到证据表明FES参与了 通过一类特殊的表面受体在细胞- 细胞和细胞-基质的相互作用，并提示FES可能的作用 在巨噬细胞中介导炎性细胞因子的释放 激活。我们发现VLA-4整合素参与巨噬细胞，a 诱导炎症介质释放的过程，导致 FeS激酶的激活。我们还确定了一些 Fes的巨噬细胞特异性底物，参与细胞 炎性细胞因子的黏附和调节。其中一个 被FeS磷酸化的底物是Crk相关底物(CAS) 另一种是新的130 kDa蛋白(P130)。CAS是一种适配蛋白 参与整合素信号传递。P130是一种仅在 造血细胞与炎症的释放有关 细胞因子。我们假设Fes参与了一种新的整合素- 调节炎症介质释放的依赖途径。 在这个提案中，我们将研究整合素的机制 参与导致FeS激酶的激活，以及如何磷酸化 其特定的底物调节炎症反应 巨噬细胞。巨噬细胞黏附受体VLA-4将通过 通过与特定抗体和与纤维连接蛋白的粘连进行交联， 以及Fes与整合素级联的功能相互作用 将会被检查。使用遗传和生物化学的组合 方法：我们将阐明Fes激酶的作用机制 活化，表征FeS底物，确定其下游 靶点，并分析Fes在黏附依赖中的作用 巨噬细胞炎症反应。最后，我们将确定 Fes激活的转录调控因子和Fes反应图谱 FES控制的基因启动子中的元件。巨噬细胞 在对微生物的免疫反应中发挥核心作用 杀瘤活性，在病理条件下，如 动脉硬化。拟议中的调查将揭露一个 此前未知的参与炎症反应的调节级联反应 巨噬细胞。这将对合理的设计产生影响。 旨在防止不适当激活的治疗方法 巨噬细胞，一个至关重要的生物医学问题。