MANIPULATING GLUCOCORTICOID RECEPTOR IN LIVE TUMOR CELLS

我的长期目标是了解 受体介导的糖皮质激素作用。 糖皮质激素是 作为免疫抑制剂和抗炎剂都很重要 药物和作为治疗工具治疗白血病， 淋巴瘤 目前，知识还远远没有完成， 类固醇敏感性肿瘤细胞的病因学和/或控制， 激素诱导肿瘤反应的分子机制 正常细胞，或者一些细胞逃避激素的途径 控制 这部分是由于传统方法的性质 类固醇受体的研究，因为他们依赖于间接的 检测策略需要形成和维持 类固醇受体复合物，通常在细胞中进行， 自由系统 该项目的目的是开发和 评估一种新的实验系统，其中分子和 将直接探讨类固醇激素作用的细胞生物学 在活细胞中。 生物探针将被引入 系统是：1）单克隆抗体，其结合到 糖皮质激素受体上的表位，并改变糖皮质激素受体的功能， 和2）免疫亲和纯化的完整受体及其 含有特定功能域的离散片段。 一 基于红细胞幻影介导方法的改进策略 将被用来“注射”大量的这些生物 活性大分子转化为糖皮质激素的大量培养物， 反应性和抗性肿瘤细胞。 如图所示， 实验系统具有显著的通用性， 使新的体内受体中和，竞争， 和替代研究，在任何可用的糖皮质激素- 应答和变异抗性细胞系。 抗体库I 希望开发也应该提供强大的新的基础 分析程序，并允许新的洞察力的结构和 特别是糖皮质激素受体的功能， 受体一般。

My long-term objective is to understand the mechanism of receptor-mediated glucocorticoid action. Glucocorticoids are important both as immunosuppressive and anti-inflammatory agents and as therapeutic tools in the treatment of leukemias and lymphomas. Currently, knowledge is far from complete regarding the etiology and/or control of steroid-sensitive tumor cells, the molecular mechanism of steroid-induced responses in neoplastic on normal cells, or the route by which some cells escape hormonal control. This is due in part to the nature of traditional methods of steriod receptor study in that they have relied on indirect detection strategies requiring the formation and maintenance of steroid-receptor complexes, and are usually performed in cell- free systems. The purpose of this project is to develop and evaluate a novel experimental system in which the molecular and cellular biology of steriod hormone action will be directly probed in living cells. The biological probes to be introduced into the system are 1) monoclonal antibodies that bind to an array of epitopes on, and modify function of the glucocorticoid receptor, and 2) the immunoaffinity-purified intact receptor and its discrete fragments containing specific functional domains. A refined strategy based on the erythrocyte ghost-mediated method will then be used to "inject" large numbers of these biologically active macromolecules into bulk cultures of glucocorticoid- responsive and resistant tumor cells. As formulated, this experimental system possesses marked versatility, and thus will make feasible novel in vivo receptor neutralization, competition, and replacement studies in any of the available glucocorticoid- responsive and variant resistant cell lines. The antibody bank I hope to develop should also provide the basis of powerful new analytical procedures and allow new insight into the structure and function of the glucocorticoid receptor in particular, and steroid receptors in general.