The Glucocorticoid Receptor as Signal Integrator: Studying All Drug Resistance

项目摘要/摘要 蛋白质位于信号通路的节点，它们的任务是整合这些信号来指导特定的 输出--根据细胞当时的需要量身定做的输出。迈尔斯·普法尔博士研究过信号集成 使用糖皮质激素受体，特别是DNA调节变构的机制基础 序列。K99/R00奖将为他提供开发系统所需的资源、时间和培训 将GR变构整合的结构和生物物理基础转化为对 导致儿童前B急性淋巴细胞白血病(B-ALL)治疗的信号异常 抵抗。这项工作将使选择性干预措施的发展成为可能，以指导变构网络在 蛋白。这一奖项将使普费尔博士能够实现以下职业发展目标：1)制定 立足于造血发育和疾病；2)学习使用初级组织；3)获得 对基因组数据集的经验，以发展未来的研究；以及4)开发一个体外系统来测试信号 整合原则。 糖皮质激素受体通过以下方式协调基因表达程序以响应细胞信号 使整个基因组中特定DNA反应元件上的调控复合体的组装成核。 在B-ALL中，针对GR的合成糖皮质激素是一种有效的治疗方法，联合应用 其他药物则能诱导细胞凋亡。然而，那些对糖皮质激素没有反应的人 面临着严峻的预测。普法尔博士假设，通过变构调节GR的信号通路 被干扰，改变受体的正常功能，阻断细胞凋亡。他将测试这一点 假设有三个目的：1)确定糖皮质激素诱导的基因表达程序、GR结合、 糖皮质激素敏感和耐药急性淋巴细胞白血病的活性染色质标记；2) 从选定的反应元件中提纯GR调节复合体并鉴定成分；以及3)鉴定 从GR的DNA结合域发出的变构导线，并确定信号如何影响这些 电线。

Project Summary/Abstract Proteins lie at the nodes of signaling pathways, and it is their task to integrate these signals to direct a specific output - one that is tailored to the needs of the cell at the time. Dr. Miles Pufall has studied signal integration using the glucocorticoid receptor (GR), in particular the mechanistic basis of allosteric regulation by DNA sequence. This K99/R00 award will provide him with the resources, time, and training to develop systems that translate the structural and biophysical basis of GR allosteric integration into an understanding of the signaling aberrations that result in childhood pre-B acute lymphoblastic leukemia (B-ALL) treatment resistance. This work will enable development of selective interventions that direct allosteric networks in the protein. This award will allow Dr. Pufall to achieve the following career development goals: 1) Develop a grounding in hematopoietic development and disease; 2) Learn to work with primary tissue; 3) Gain experience with genomic data sets to develop future studies; and 4) Develop an in vitro system to test signal integration principles. The glucocorticoid receptor orchestrates a program of gene expression in response to cellular signals by nucleating the assembly of regulatory complexes at specific DNA response elements throughout the genome. In B-ALL, synthetic glucocorticoids directed against GR are an effective treatment, which in combination with other drugs, work to induce an apoptotic program. However those who do not respond to glucocorticoids face a grim prognosis. Dr. Pufall hypothesizes that signaling pathways that allosterically regulate GR have been disrupted, changing the normal function of the receptor, and blocking apoptosis. He will test this hypothesis in three aims: 1) Identify changes in glucocorticoid induced gene expression programs, GR binding, and marks for active chromatin in glucocorticoid sensitive and resistant acute lymphoblastic leukemia; 2) Purify GR regulatory complexes from select response elements and identify components; and 3) Identify allosteric wires emanating from the DNA binding domain of GR, and determine how signals impinge on these wires.