The glucocorticoid receptor as signal integrator: studying ALL drug resistance

作为信号整合器的糖皮质激素受体：研究 ALL 耐药性

• 批准号: 7868075
• 负责人: MILES A PUFALL
• 金额: $ 11.54万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-21 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7868075
• 关键词: Acute Lymphocytic Leukemia; Adverse effects; Allosteric Regulation; Apoptosis; Apoptotic; Asthma; Autoimmune Diseases; Award; B-Cell Acute Lymphoblastic Leukemia; Binding Sites; Biochemical; Biology; Calorimetry; Cell Line; Cells; Characteristics; Childhood; Chromatin; Communication; Complex; Computing Methodologies; Cues; DNA; DNA Binding; DNA Binding Domain; DNA Sequence; Data Set; Development; Diabetes Mellitus; Disease; Drug resistance; Drug usage; Face; Future; Gene Expression; Genes; Genetic Transcription; Genome; Genomics; Glucocorticoid Receptor; Glucocorticoid-induced apoptosis; Glucocorticoids; Goals; Health; Hematopoietic; Histone H3; Hormones; Hypertension; In Vitro; Intervention; Learning; Lysine; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Measures; Messenger RNA; Methods; Modeling; Modification; Nuclear Extract; Nuclear Magnetic Resonance; Organ Transplantation; Osteoporosis; Outcome; Output; Pharmaceutical Preparations; Pre-B Acute Lymphoblastic Leukemia; Prednisone; Property; Proteins; Reading; Resistance; Resources; Response Elements; Screening procedure; Signal Pathway; Signal Transduction; Specific qualifier value; Structural Models; System; Testing; Thermodynamics; Time; Tissues; Training; Translating; Work; base; career development; cell type; effective therapy; experience; genome-wide; glucocorticoid-induced orphan receptor; hormone resistance; human tissue; outcome forecast; programs; public health relevance; receptor; receptor binding; receptor function; response; transcription factor

DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE: Glucocorticoid hormones act through their receptor, GR, to regulate gene expression. Drugs that activate GR are used for a wide range of conditions, from asthma to organ transplants to cancer, such as acute lymphoblastic leukemia. However, these drugs are not always effective, and they also have serious side effects, including diabetes, osteoporosis, and hypertension. Thus, understanding how GR integrates and responds to cellular cues has important implications for health, and for detecting, treating and curing disease.

描述（由申请人提供）：蛋白质位于信号传导途径的节点，它们的任务是整合这些信号以指导特定的输出-一种适合细胞当时需要的输出。Miles Pufall博士使用糖皮质激素受体（GR）研究了信号整合，特别是DNA序列变构调节的机制基础。这个K99/R 00奖将为他提供资源，时间和培训，以开发系统，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）治疗抗性的信号畸变的理解。这项工作将使选择性干预的发展，直接在蛋白质的变构网络。该奖项将使Pufall博士能够实现以下职业发展目标：1）在造血发育和疾病方面奠定基础; 2）学习使用原代组织; 3）获得基因组数据集的经验，以开发未来的研究; 4）开发体外系统以测试信号整合原理。 糖皮质激素受体通过在整个基因组的特定DNA反应元件处成核调节复合物的组装来协调响应细胞信号的基因表达程序。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，与其他药物联合使用，可诱导凋亡程序。然而，那些对糖皮质激素没有反应的人面临着严峻的预后。Pufall博士推测，变构调节GR的信号通路已被破坏，改变了受体的正常功能，并阻断了细胞凋亡。他将在三个目标中检验这一假设：1）确定糖皮质激素诱导的基因表达程序、GR结合以及糖皮质激素敏感和耐药急性淋巴细胞白血病中活性染色质的标记的变化; 2）从选择的反应元件中纯化GR调节复合物并鉴定组分; 3）鉴定从GR的DNA结合域发出的变构丝，并确定信号如何冲击这些丝。 公共卫生相关性：糖皮质激素通过其受体GR调节基因表达。激活GR的药物用于广泛的疾病，从哮喘到器官移植到癌症，如急性淋巴细胞白血病。然而，这些药物并不总是有效的，它们也有严重的副作用，包括糖尿病，骨质疏松症和高血压。因此，了解GR如何整合和响应细胞信号对健康以及检测，治疗和治愈疾病具有重要意义。