Developmental control of chromatin states in cancer

癌症中染色质状态的发育控制

• 批准号: 10567242
• 负责人: Ian J Davis
• 金额: $ 41.15万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2027-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10567242
• 关键词: Acceleration; Adolescent; Adolescent and Young Adult; Binding; Biochemical; Biological Models; Bone Tissue; CRISPR interference; Cell Differentiation process; Cell Proliferation; Cell physiology; Cells; Characteristics; Child; Chimeric Proteins; Chromatin; Chromosomal Rearrangement; Chromosomal translocation; Common Neoplasm; Dependence; Development; ETS Domain; EWS-FLI1 fusion protein; EWSR1 gene; Enabling Factors; Enhancers; Environment; Epigenetic Process; Ewings sarcoma; FLI1 gene; Family member; Fostering; Fusion Oncogene Proteins; Generations; Genes; Genetic; Genetic Transcription; Genomics; Growth; Heterogeneity; Histologic; Histones; Human; Impact evaluation; In Vitro; Label; Link; Malignant Bone Neoplasm; Malignant Childhood Neoplasm; Malignant Neoplasms; Mass Spectrum Analysis; Mediator; Mesenchymal Cell Neoplasm; Mesenchymal Differentiation; Mesenchymal Stem Cells; Microsatellite Repeats; Oncogenic; Oncoproteins; Operative Surgical Procedures; PAX7 gene; Patients; Pluripotent Stem Cells; Post-Translational Protein Processing; Process; Proliferating; Proteins; Proteomics; RNA; Radiation; Regulation; Regulatory Pathway; Research Personnel; Site; Testing; Untranslated RNA; Variant; Work; cancer cell; chemotherapy; cofactor; embryonic stem cell; epigenomics; histone modification; hybrid gene; induced pluripotent stem cell; insight; neoplastic cell; new therapeutic target; novel; novel strategies; permissiveness; protein function; recruit; sarcoma; small hairpin RNA; soft tissue; stem cell differentiation; stem cell model; stem cells; transcription factor; transcriptome; transcriptomics; tumor; young adult

SUMMARY Ewing sarcoma, a cancer of the bone and soft tissue of children and young adults, remains a highly lethal cancer despite the use of aggressive chemotherapy, radiation, and surgery. The tumor is dependent on the development of a hybrid gene that brings together parts of two different genes, EWSR1 and FLI1, through chromosomal translocation or chromoplexy. The resulting EWS-FLI1 fusion oncoprotein acts as a transcriptional and chromatin regulator. Building from the work of our lab and others demonstrating that EWS-FLI1 gains neomorphic activity to regulates chromatin state at microsatellite coopted to become enhancers in tumor cells. However, we have shown that the activity and genomic targeting of EWS-FLI1 is influenced by the underlying epigenomic state of the cell. We demonstrated that primary and in vitro differentiated mesenchymal stem cells offer a chromatin state similar to that of Ewing sarcoma. We hypothesize that during stem cell differentiation a unique permissive chromatin state develops that enables EWS-FLI1. Further, we hypothesize that this permissive state is made up of chromatin regulators, characteristically modified histones and specific RNAs. In this project we will employ cancer cell (Aim 1) and stem cell developmental approaches (Aim 2) to identify the protein and RNA interactors and the posttranslational modifications of histones that create a favorable environment. We predict that critical features will be shared between both model systems. We will test the impact of these factors by evaluating the activity of EWS-FLI1 on chromatin states and transcription when these factors are modulated. The generation of Ewing sarcoma patient derived stem cells will enable us to evaluate the impact of EWS-FLI1 across the process of cellular differentiation. Integrated single cell analytics of chromatin accessibility and the transcriptome will enable direct evaluation of the impact of EWS-FLI1 on chromatin and, reciprocally, chromatin on EWS-FLI1. We will specifically study the biochemical interaction and functional relationship between EWS-FLI1 and the transcriptional regulator PAX7, one of the interactors identified during pilot experimentation.

概括 尤因肉瘤是儿童和年轻人的骨骼和软组织的癌症，仍然是一个高度致命的癌症 尽管使用了积极的化学疗法，放射线和手术。肿瘤取决于发育 通过染色体将两个不同基因EWSR1和FLI1的一部分的杂种基因汇集在一起 易位或铬流体。所得的EWS-FLI1融合癌蛋白充当转录和染色质 监管机构。从我们的实验室和其他人的工作中构建，表明EWS-FLI1获得了新态活动 调节在微卫星处的染色质状态，以成为肿瘤细胞的增强子。但是，我们有 表明EWS-FLI1的活性和基因组靶向受到潜在的表观基因组状态的影响 细胞。我们证明了原发性和体外分化的间充质干细胞提供染色质状态 类似于尤因肉瘤。我们假设在干细胞分化期间独特的允许性 染色质状态发展起来可以启用EWS-FLI1。此外，我们假设这种允许的状态已成为 在染色质调节剂中，特征性修饰的组蛋白和特定的RNA。在这个项目中，我们将 采用癌细胞（AIM 1）和干细胞发育方法（AIM 2）来鉴定蛋白质和RNA 相互作用的人和创造有利环境的组蛋白的翻译后修饰。我们预测 两个模型系统之间将共享关键功能。我们将通过 在调节这些因素时，评估EWS-FLI1对染色质状态和转录的活性。 Ewing肉瘤患者得出的干细胞的产生将使我们能够评估EWS-FLI1的影响 在整个细胞分化过程中。染色质可及性的综合单细胞分析和 转录组将直接评估EWS-FLI1对染色质的影响，并相互评估染色质 在EWS-FLI1上。我们将专门研究生化相互作用和功能关系 EWS-FLI1和转录调节器PAX7，这是试点期间确定的交互者之一 实验。