MEKK2 KO DEFINES HEMATOPOIETIC CELL SIGNAL FUNCTION

该项目的目标是利用MEK激酶2的靶向基因中断(MEKK2)来确定其在调节造血细胞的信号通路和基因表达中的作用。初步数据显示，在ES细胞来源的肥大细胞中，MEKK2的丢失导致c-Kit和FcepsilonRI结扎反应中JNK活性的丧失，并且刺激细胞中所选择的细胞因子的产生显著减少。在被T细胞/抗原提呈细胞结合形成激活的T细胞中，MEKK2迅速移位到结合界面。转染酶失活的MEKK2可阻止结合物的形成，并轻微抑制IL-4的产生。本项目的具体目的是建立MEKK2基因敲除小鼠，确定基因敲除表型，并从小鼠中收集肥大细胞和T细胞，以研究MEKK2基因缺失对信号通路、细胞增殖和细胞因子产生的影响。将使用新技术，如ES细胞的体外分化和微芯片DNA阵列技术。这些研究的结果将对理解MEKK2的S在信号通路中的作用以及它在细胞因子产生和免疫功能中的调控作用产生影响。他们还可能在癌症研究领域有意义，因为在T细胞中转染MEKK2可以影响细胞周期的速度。最终，这些研究可能导致以影响MEKK2活性为目标的新疗法的开发，以治疗癌症或调节免疫功能。作为一名儿科血液学家和肿瘤学家，这个项目将在教我细胞信号、细胞生长控制和免疫学等领域的研究方法方面发挥无价的作用，所有这些领域都与我的领域密切相关。我的长期目标是将这种增长和免疫学应用到与我的领域密切相关的所有领域。我的长期目标是应用这些知识来更好地了解导致信号通路失调并导致儿童癌症的细胞事件，从而设计出有效的、有针对性的治疗方法。加里·约翰逊的实验室是学习这些技能的理想环境。

The goal of this project is to use targeted gene disruption of MEK kinase 2 (MEKK2) to determine its role in regulation of signaling pathways and of gene expression in hematopoietic cells. Preliminary date has shown that loss of MEKK2 in ES cell derived mast cells results in loss JNK activation in response to c-Kit and FcepsilonRI ligation and there is significantly decreased production of selected cytokines in the stimulated cells. In T cells with activation by T cell/antigen presenting cell conjugate formation, MEKK2 rapidly translocates to the conjugate interface. Transfected kinase inactive MEKK2 prevents conjugate formation and slightly inhibits IL-4 production. Specific aims of this project are to generate MEKK2 knockout mice, to determine the knockout phenotype, and to collect mast cells and T cells from the mice to study effects of loss of MEKK2 on signaling pathways, cell proliferation, and cytokine production. Novel techniques, such as in vitro differentiation of ES cells and microchip DNA array technology, will be used. Results of these studies will have an impact on the understanding of MEKK2's role in signaling pathways as well as its role in control of cytokine production and immune function. They could also have implications in the field of cancer research, since transfected MEKK2 in T cells can affect rate of cell cycling. Ultimately, these studies may lead to the development of novel therapies targeted at influencing MEKK2 activity to treat cancers or to modulate immune function. As a pediatric hematologist, oncologist, this project will be invaluable in teaching me research methods in the areas of cell signaling, control of cell growth, and immunology, all areas intimately involved with my field. My long term goal is to apply this growth, and immunology, all areas intimately involved with my field. My long term goal is to apply this knowledge to gain a better understanding of cellular events leading to dysregulation of signaling pathways and resulting in cancer in children, so that effective, targeted therapies can be devised. Gary Johnson's lab is an ideal environment in which to learn these skills.