EWS/FLI Regulates Transcriptional Activation in Ewing Sarcoma via Length Dependent GGAA Microsatellites

EWS/FLI 通过长度依赖性 GGAA 微卫星调节尤文肉瘤的转录激活

• 批准号: 9312656
• 负责人: Kirsten Johnson
• 金额: $ 4.9万
• 依托单位: RESEARCH INST NATIONWIDE CHILDREN'S HOSP
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-23 至 2019-08-22
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9312656
• 关键词: Affect; Affinity; Binding; Binding Sites; Biochemical; Biochemistry; Biological Assay; Biological Markers; CRISPR/Cas technology; Cell Line; Cells; Childhood; Childhood Cancer Treatment; Chromosomal translocation; Clinical; Clinical Data; Crime; DNA; DNA Binding; DNA Binding Domain; DNA Sequence Alteration; Development; Disease; Disease susceptibility; EMSA; Enhancers; Ewings sarcoma; Family member; Fluorescence Anisotropy; Fusion Oncogene Proteins; Gel; Gene Expression; Gene Targeting; Genetic; Genetic Transcription; Goals; Growth; Heritability; Hydrogels; In Vitro; Inherited; Junk DNA; Knowledge; Length; Link; Malignant Bone Neoplasm; Malignant Childhood Neoplasm; Malignant Neoplasms; Measures; Mediating; Microsatellite Repeats; Modeling; Molecular; Molecular and Cellular Biology; NR0B1 gene; Neoplasm Metastasis; Oligonucleotides; Oncogenes; Oncogenic; Oncoproteins; Operative Surgical Procedures; Outcome; Pathogenesis; Patient Care; Patients; Polymers; Predisposition; Process; Promoter Regions; Property; Proteins; Recombinant Proteins; Recruitment Activity; Repression; Response Elements; Reverse Transcriptase Polymerase Chain Reaction; Role; Site; Spottings; Survival Rate; System; Testing; Transcription Process; Transcriptional Activation; Transcriptional Activation Domain; Transcriptional Regulation; Translating; Untranslated RNA; Western Blotting; Work; biomarker development; bone; chemotherapy; clinically relevant; effective therapy; experimental study; improved; insight; leukemia; mutant; novel; promoter; stoichiometry; synthetic construct; therapeutic development; therapeutic target; transcription factor; tumor progression; tumorigenesis

Project Summary/Abstract Our long-term goal in this project is to elucidate the molecular and cellular biology for Ewing Sarcoma development and translate this knowledge into advances for pediatric cancer. Ewing sarcoma, the second most common pediatric malignant bone tumor, is characterized by a chromosomal translocation t(11;22). The resulting fusion oncoprotein EWS/FLI is an aberrant transcription factor necessary for oncogenic transformation in Ewing cells. EWS acts as an amino-terminal transcriptional activation domain linked to the carboxy-terminal DNA-binding domain of its ETS family member partner in crime, FLI. Microsatellite repeats characterized by the motif GGAA serve as binding sites for EWS/FLI within the promoters of upregulated target genes in Ewing Sarcoma. Previously thought of as “junk DNA,” these polymorphic microsatellites serve as response elements for EWS/FLI DNA binding with interesting genetic correlations and possible clinical implications. Though EWS/FLI both activates and represses numerous target genes directly and indirectly, how EWS/FLI distinguishes whether to up or down regulate gene expression remains undetermined. Therefore, the experiments proposed herein aim to test the hypothesis that EWS/FLI induces oncogenic transformation and regulation of transcriptional activation via GGAA microsatellites whose length dictates both binding and effector function. Thus our aims are: 1) Determine to what extent GGAA microsatellites function as enhancer response elements and 2) Determine the role of GGAA motif length in transcriptional activation in Ewing Sarcoma. This project is an important approach to pediatric cancer because it provides a model of understanding oncogenic transcriptional regulation necessary for biomarker and therapeutic development to improve patient care and overall survival.

项目概要/摘要 我们在这个项目中的长期目标是阐明尤文肉瘤的分子和细胞生物学 开发并将这些知识转化为儿科癌症的进展。尤因肉瘤，第二个 最常见的儿童恶性骨肿瘤，其特征是染色体易位 t(11;22)。这 由此产生的融合癌蛋白 EWS/FLI 是致癌所必需的异常转录因子 尤文细胞的转化。 EWS 作为氨基末端转录激活结构域，与 其 ETS 家族成员犯罪伙伴 FLI 的羧基末端 DNA 结合域。微卫星重复序列 以 GGAA 基序为特征，作为上调靶标启动子内 EWS/FLI 的结合位点 尤文肉瘤的基因。这些多态性微卫星以前被认为是“垃圾DNA” EWS/FLI DNA 结合的反应元件具有有趣的遗传相关性和可能的​​临床 影响。尽管 EWS/FLI 既直接又间接地激活和抑制众多靶基因，但如何 EWS/FLI 区分是否上调或下调基因表达仍不确定。因此， 本文提出的实验旨在检验 EWS/FLI 诱导致癌转化的假设 通过 GGAA 微卫星调节转录激活，其长度决定结合和效应子 功能。因此，我们的目标是：1) 确定 GGAA 微卫星在多大程度上发挥增强子反应的作用 元件和 2) 确定 GGAA 基序长度在尤文肉瘤转录激活中的作用。这 该项目是治疗儿科癌症的一个重要方法，因为它提供了一个理解致癌因素的模型 生物标志物和治疗开发所必需的转录调控，以改善患者护理和 总体生存率。