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

基本信息

批准号：
8075410
负责人：
MILES A PUFALL
金额：
$ 11.79万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-05-21 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8075410
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 RNA 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

项目摘要

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）：蛋白质位于信号通路的节点，它们的任务是整合这些信号以指导特定的输出——根据细胞当时的需要定制的输出。 Miles Pufall 博士研究了糖皮质激素受体 (GR) 的信号整合，特别是 DNA 序列变构调节的机制基础。该 K99/R00 奖项将为他提供资源、时间和培训来开发系统，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 期急性淋巴细胞白血病 (B-全部）治疗抵抗。这项工作将有助于开发指导蛋白质变构网络的选择性干预措施。该奖项将使 Pufall 博士能够实现以下职业发展目标： 1) 奠定造血发育和疾病方面的基础； 2）学习使用原代组织； 3）获得基因组数据集的经验以开展未来的研究； 4) 开发体外系统来测试信号集成原理。糖皮质激素受体通过在整个基因组的特定 DNA 响应元件处使调节复合物的组装成核，来协调响应细胞信号的基因表达程序。对于 B-ALL，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细胞凋亡程序。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，从而改变了受体的正常功能，并阻止了细胞凋亡。他将在三个目标上检验这一假设：1）确定糖皮质激素诱导的基因表达程序、GR 结合的变化，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标记的变化； 2) 从选定的响应元素中纯化GR调控复合物并识别成分； 3) 识别从 GR 的 DNA 结合域发出的变构线，并确定信号如何撞击这些线。公共卫生相关性：糖皮质激素通过其受体 GR 发挥作用，调节基因表达。激活 GR 的药物用于治疗多种疾病，从哮喘到器官移植，再到癌症（如急性淋巴细胞白血病）。然而，这些药物并不总是有效，而且还具有严重的副作用，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于健康以及检测、治疗和治愈疾病具有重要意义。