Phase Separation-involved Roles of Mediator Complex in Myc-amplified Cancer

介导复合物在 Myc 扩增癌症中的相分离作用

• 批准号: 10590572
• 负责人: CHUNYU JIN
• 金额: $ 18.58万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-11 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10590572
• 关键词: Affinity; Automobile Driving; Binding; Biological; Cancer Biology; Cancer Patient; Cell Survival; Cells; ChIP-seq; Competitive Binding; Complement; Complex; DNA Damage; Data; Dissociation; Drug Targeting; Enhancers; Event; Exhibits; Frequencies; Gene Activation; Gene Amplification; Gene Expression; Generations; Genes; Genetic; Genetic Transcription; Genomic Instability; Glioblastoma; Goals; Growth; Head; In Vitro; Individual; Interruption; Knock-out; Link; MED15; MYC Gene Amplification; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediator; Molecular; Oncogenic; Pharmaceutical Preparations; Phase; Phenotype; Phosphotransferases; Physical condensation; Physiological; Play; Proliferating; Property; Protein Complex Subunit; Proteins; Proto-Oncogene Proteins c-myc; RNA Polymerase II; Reagent; Regulation; Role; Signal Transduction; Tail; Testing; Therapeutic; Transcription Alteration; Transcriptional Activation; Transcriptional Regulation; Work; cancer cell; cancer type; carcinogenesis; drug development; extracellular; histone modification; in vivo; insight; mouse model; overexpression; programs; promoter; protein complex; protein degradation; rapid growth; replication stress; response; screening; small molecule; therapeutic target; transcription factor; tumor progression

ABSTRACT High frequency of Myc gene amplification is found in many types of cancer, driving the oncogenic activity in cancer progression. Here I have noticed that the gene encoding one component of the transcription machinery mediator complex(MED) protein, MED30, is significantly co-amplified with Myc, and thus might complement the effects of enhanced Myc transcription activity. This correlation suggests a potentially useful adjunct therapeutic target in Myc-amplified cancer. MED contains thirty components and regulates gene expression by promoting assembly of the transcription preinitiation complex (PIC) and elongation. My preliminary data has revealed the unequal structural and functional importance of individual component of MED, by which MED30, located in Tail module, is required for assemble of at least the Tail and Middle MED modules, while, in contrast, another tail protein MED15 is not required. Mechanistically, 1,6-hexanediol-disrupted phase separation play a critical role in it. MED30 depletion leads to protein degradation of many other MED components, while MED15 does not. ChIP- seq reveals MED is widely enriched in enhancer and active transcribed gene promoters, and regulates target genes that represents a cancer-associated signature. MED works as a rapid transcription switch in response to extracellular signals. Therefore, I propose to rigorously test the molecular basis for MED30 function as one of the core components of MED, linked to cancer exhibiting Myc amplification, which will be the basis for competitive binding-based drug targeting, and screening for small molecules that can interrupt mediator complex by high affinity to MED30. I will further explore the molecular mechanism of MED involved transcription regulation in synergistic with Myc, and investigate the in vivo effect of MED30 inhibition in many cancers, including pancreatic cancer and a glioblastoma mouse model. This study will simultaneously extend our understanding of the potential cooperative component of Myc-induced cancer, investigate the roles of phase separation in these biological events, particularly on the aspect of protein stability regulation important for Mediator function, as well as how the cells tune their transcriptional program through enhanced MED30 and Myc TF transcription activity. I will focus on this overlooked and apparently common event in cancer biology, with the goal of providing new insights into transcription inhibitory drug development and protein degradation-oriented drug targeting strategies.

摘要 在许多类型的癌症中发现了高频率的Myc基因扩增，从而驱动了肿瘤细胞中的致癌活性。 癌症进展在这里我注意到，编码转录机制的一个组成部分的基因 介体复合物（MED）蛋白MED 30与Myc显著共扩增，因此可能与Myc互补。 增强Myc转录活性的影响。这种相关性表明了一种潜在的有用的辅助治疗方法 靶向Myc扩增的癌症。MED含有30种成分，通过促进基因表达来调节基因表达。 转录前起始复合体（PIC）的组装和延伸。我的初步数据显示 MED各组分的结构和功能重要性不相等，因此MED 30位于尾 至少尾部和中部MED模块的组装需要另一个尾部模块 不需要MED 15蛋白。从机理上讲，1，6-己二醇破坏的相分离在 了MED 30消耗导致许多其他MED组分的蛋白质降解，而MED 15不会。ChIP- seq揭示MED广泛富集增强子和活性转录基因启动子，并调节靶点， 代表癌症相关特征的基因MED作为一种快速转录开关， 细胞外信号因此，我建议严格测试MED 30功能的分子基础， MED的核心成分，与表现出Myc扩增的癌症有关，这将是 基于竞争性结合的药物靶向，以及筛选可以中断介体复合物的小分子 对MED 30的高亲和力。本文将进一步探讨MED参与转录调控的分子机制 协同Myc，并研究MED 30抑制在许多癌症中的体内作用，包括 胰腺癌和胶质母细胞瘤小鼠模型。这项研究将同时扩大我们对 Myc诱导的癌症的潜在合作成分，研究相分离在这些过程中的作用。 生物学事件，特别是对介体功能重要的蛋白质稳定性调节方面，以及 细胞如何通过增强MED 30和Myc TF转录活性来调节其转录程序。 我将集中讨论这一被忽视的，显然是癌症生物学中常见的事件，目的是提供新的 深入了解转录抑制药物开发和蛋白质降解导向的药物靶向策略。