MEKK2 KO DEFINES HEMATOPOIETIC CELL SIGNAL FUNCTION

本项目的目标是使用靶向的MEK激酶2（MEKK 2）的基因破坏，以确定其在造血细胞中的信号通路和基因表达的调节中的作用。初步数据显示，ES细胞衍生的肥大细胞中MEKK 2的损失导致响应于c-Kit和Fc ε RI连接的JNK活化的损失，并且在刺激的细胞中所选细胞因子的产生显著降低。在通过T细胞/抗原呈递细胞缀合物形成而活化的T细胞中，MEKK 2迅速易位至缀合物界面。转染的激酶失活MEKK 2阻止缀合物形成并轻微抑制IL-4产生。该项目的具体目标是产生MEKK 2敲除小鼠，以确定敲除表型，并从小鼠中收集肥大细胞和T细胞，以研究MEKK 2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MEKK 2在信号通路中的作用以及其在控制细胞因子产生和免疫功能中的作用产生影响。它们也可能在癌症研究领域产生影响，因为T细胞中转染的MEKK 2可以影响细胞周期的速率。 最终，这些研究可能会导致开发新的治疗方法，以影响MEKK 2活性来治疗癌症或调节免疫功能。作为一名儿科血液学家，肿瘤学家，这个项目将是非常宝贵的，在教我在细胞信号传导，细胞生长控制和免疫学领域的研究方法，所有领域密切涉及我的领域。我的长期目标是将这种增长和免疫学应用于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，以便设计有效的靶向治疗。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.