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Regulation of Stat Function in Breast Cancer

乳腺癌中 Stat 功能的调节

基本信息

批准号：
7224178
负责人：
Charles V Clevenger
金额：
$ 24.6万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-07-01 至 2009-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7224178
关键词：
Binding Biochemical Biology Breast Breast Cancer Cell Cancer cell line Cell Line Characteristics Complement Complex DNA Binding Data Differentiation and Growth Disease Elements Endocytosis Pathway Family Family member Functional disorder Gene Expression Genetic Transcription Genomics Growth Half-Life Hormones Human In Vitro Laboratories Ligation MYB gene Malignant Neoplasms Mammary gland Maps Mediating Methodology Molecular Mus Mutagenesis Neurosecretory Systems Nuclear Numbers Pathogenesis Peptides Peptidylprolyl Isomerase Phosphorylation Prolactin Protein Overexpression Protein Tyrosine Kinase Proteins Proto-Oncogene Proteins c-myb RNA Receptor Activation Regulation Research Personnel Signal Transduction Site Small Interfering RNA Technology Testing Tissues Transcription factor genes Transgenic Organisms Ubiquitin Xenograft Model Yeasts base chromatin immunoprecipitation cyclophilin B genetic regulatory protein immunosuppressed in vivo inhibitor/antagonist malignant breast neoplasm mutant non-genomic novel novel therapeutics programs promoter protein function protein protein interaction receptor receptor mediated endocytosis transcription factor ubiquitin-protein ligase yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): The Stat family of transcription factors is necessary for the growth and differentiation of mammary tissues. In mammary tissues, a significant means of Stat activation is the binding of the neuroendocrine hormone prolactin (PRL) to its receptor, the PRLr. PRL/PRLr binding results in the activation of receptor-associated signaling networks, such as the Jak/Stat pathway and the endocytosis and nuclear retrotransport of a complex between PRL and the prolyl isomerase cyclophilin B (CypB). The intranuclear PRL/CypB complex regulates Stat5-mediated gene expression and DNA binding. Using multiple approaches including yeast two-hybrid and transcription factor array analysis, we have demonstrated that the PRL/CypB complex regulates the interaction of Stat5 with multiple intranuclear proteins including the peptide inhibitor of activated Stat, PIAS3, the small ubiquitin-related modifier, SUMO2, and the transcription factor and proto-oncogene, c-Myb. It is the central hypothesis of this proposal that the temporal and spatial interaction of Stat5 with these proteins, as modulated by the PRL/CypB complex, regulates Stat5 function. This hypothesis will be tested by three specific aims using breast cancer and PRL-responsive cell lines both in vitro and in vivo. First, the structural basis for Stat5/PIAS3 interaction and the functional significance of this interaction will be mapped using complementary mutagenesis/overexpression and knockdown strategies. Second, the functional consequences of Stat5 sumolyation will be evaluated by cellular and biochemical approaches. Third, the association between Stat5 and co-activators/repressors, such as c-Myb will be assessed by targeted mutagenesis, small-interfering RNA (SiRNA), and chromatin immunoprecipitation (ChlP)-based methodologies. Given that our preliminary evidence indicates that the manipulation of these interactions results in the alteration of Stat5-mediated gene expression and the survival of breast cancer cells, our proposed studies are directly relevant to a better understanding of the pathophysiology of: human breast cancer, and as such, may provide the basis for novel therapeutic strategies aimed at modifying Stat function in this disease.

描述(申请人提供)：Stat转录因子家族是乳腺组织生长和分化所必需的。在乳腺组织中，STAT激活的一个重要途径是神经内分泌激素催乳素(PRL)与其受体PRLr的结合。PRL/PRLr结合导致受体相关信号网络的激活，如Jak/Stat途径，以及PRL和Pro异构酶亲环素B(CypB)之间的复合体的内吞和核反向转运。核内PRL/CypB复合体调节Stat5介导的基因表达和DNA结合。利用酵母双杂交和转录因子阵列分析等多种方法，我们已经证明了PRL/CypB复合体调节Stat5与多种核内蛋白的相互作用，包括激活STAT的多肽抑制物PIAS3，小泛素相关修饰物SUMO2，以及转录因子和原癌基因c-Myb。这一假设的中心假设是，在PRL/CypB复合体的调控下，Stat5与这些蛋白的时空相互作用调节着Stat5的功能。这一假设将通过三个特定的目标在体外和体内通过乳腺癌和PRL反应细胞株进行验证。首先，将使用互补突变/过表达和敲除策略来定位Stat5/PIAS3相互作用的结构基础和这种相互作用的功能意义。其次，将通过细胞和生化方法评估Stat5总和的功能后果。第三，将通过靶向突变、小干扰RNA(SiRNA)和染色质免疫沉淀(ChlP)的方法来评估Stat5和共激活/抑制因子之间的关联，如c-Myb。鉴于我们的初步证据表明，这些相互作用的操纵导致STAT5介导的基因表达的改变和乳腺癌细胞的存活，我们拟议的研究直接与更好地理解人类乳腺癌的病理生理相关，因此，可能为旨在改变这种疾病的STAT功能的新的治疗策略提供基础。