Determining the significance of the N-MYC-WDR5 interaction in neuroblastoma.

确定 N-MYC-WDR5 相互作用在神经母细胞瘤中的重要性。

• 批准号: 10290439
• 负责人: April M. Weissmiller
• 金额: $ 37.98万
• 依托单位: MIDDLE TENNESSEE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10290439
• 关键词: Acute; Age; Agreement; Anchorage-Independent Growth; Area; Automobile Driving; Binding; Biologic Characteristic; Biological Assay; Biology; Biomass; Cell Line; Cell Proliferation; Chemicals; Child; Chromatin; Clinic; Collaborations; Combined Modality Therapy; DNA Sequence; Data; Diagnosis; Disease; Distant Metastasis; Event; Family; Family member; Foundations; Gap Junctions; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Genome; Genomic approach; Goals; Histologic; Human; Impairment; Infant; Intervention; Link; Location; Lymphoma; MYC Family Protein; MYC gene; Maintenance; Malignant Childhood Neoplasm; Malignant Neoplasms; Measures; Modeling; Mus; N-myc Gene; Names; Neuroblastoma; Oncogenic; Oncoproteins; Primary Neoplasm; Protein Biosynthesis; Publishing; Research Proposals; Resolution; Route; Site; Solid; Solid Neoplasm; Surface; Survival Rate; Testing; Therapeutic; Traction; Transcription Process; Translations; Work; angiogenesis; base; c-myc Genes; cancer therapy; chromatin immunoprecipitation; combat; design; disorder risk; gene synthesis; genetic approach; genome-wide; high risk; in vivo; member; migration; mouse model; mutant; neuroblastoma cell; new therapeutic target; next generation sequencing; programs; recruit; risk stratification; small molecule; small molecule inhibitor; transcription factor; transcriptomics; tumor; tumorigenesis

PROJECT SUMMARY/ABSTRACT Neuroblastoma (NB) is the most common extracranial solid tumor found in infants, with almost all cases being diagnosed by the age of five. The most severe cases of neuroblastoma are linked to amplification of the N-MYC oncogene, which occurs in ~20% of all NB. N-MYC amplification is associated with ~50% overall survival despite aggressive multimodal therapies, highlighting the need for new targeted therapies to combat this pediatric cancer. N-MYC, like any MYC protein, must be able to recognize and bind chromatin in order to activate its oncogenic potential. This project is built on the premise that an essential co-factor for MYC proteins called WDR5 acts to recruit N-MYC to chromatin at genes that work to maintain neuroblastoma function, and that targeting the N-MYC-WDR5 interaction can serve as a focal point for anti- N-MYC based therapies. Support for this hypothesis comes from published data showing that WDR5 is a conserved regulator of protein synthesis genes across multiple cellular contexts where WDR5 binds and regulates these loci through its so-called “WIN”-site. In addition, the MYC family member, c-MYC, requires WDR5 to bind genes associated with biomass accumulation and translation, and the c-MYC-WDR5 interaction is essential for tumor maintenance in lymphoma mouse models. In an N-MYC amplified NB cell line, preliminary data reveal that targeting the WIN-site of WDR5 using small molecule inhibitors results in a genome-wide decrease in WDR5 binding, with a subsequent loss of N-MYC binding at specific N-MYC- WDR5 co-bound genes. And, importantly, disrupting the N-MYC-WDR5 interaction using genetic mutants impairs the ability of N-MYC to drive anchorage-independent growth, providing evidence that there are essential tumor functions tied to N-MYC that require interaction with WDR5. Together these data provide a solid foundation for the notion that N-MYC requires WDR5 to bind and regulate N-MYC-WDR5 co-bound genes that are important for driving N-MYC specific activities. The goal of this project is to interrogate the N- MYC-WDR5 interaction in N-MYC amplified neuroblastoma cell lines and determine the influence that WDR5 has on N-MYC driven transcriptional processes and neuroblastoma function using genetic and chemical perturbations. Specific Aim 1 will employ genetic and genomic approaches to identify the genes that are bound by N-MYC and WDR5, and determine at which genes the recruitment of N-MYC is dependent on the N-MYC-WDR5 interaction. Specific Aim 2 will combine high-resolution transcriptomic analyses with cellular and in vivo functional assays to challenge the significance of the N-MYC-WDR5 interaction. At the completion of these studies we will have identified the precise genes in which N-MYC binding and transcription is regulated by the N-MYC-WDR5 interaction, and have directly challenged the significance of the N-MYC-WDR5 interactions on multiple facets of NB tumorigenesis.

项目概要/摘要 神经母细胞瘤（NB）是婴儿中最常见的颅外实体瘤，几乎所有病例 五岁时被诊断。最严重的神经母细胞瘤病例与神经母细胞瘤的扩增有关 N-MYC 癌基因，约 20% 的 NB 中都有该基因。 N-MYC 扩增与约 50% 相关 尽管采取积极的多模式治疗，但总体生存率仍然较高，这凸显了需要新的靶向治疗 对抗这种小儿癌症。 N-MYC 与任何 MYC 蛋白一样，必须能够识别并结合染色质 以激活其致癌潜力。该项目建立的前提是一个重要的辅助因素 称为 WDR5 的 MYC 蛋白可将 N-MYC 募集到基因处的染色质，从而维持 神经母细胞瘤功能，并且靶向 N-MYC-WDR5 相互作用可以作为抗神经母细胞瘤的焦点 基于 N-MYC 的疗法。已发表的数据支持这一假设，表明 WDR5 是一种 WDR5 结合的多种细胞环境中蛋白质合成基因的保守调节因子 通过其所谓的“WIN”位点调节这些基因座。此外，MYC 家族成员 c-MYC 需要 WDR5 结合与生物量积累和翻译相关的基因，以及 c-MYC-WDR5 相互作用对于淋巴瘤小鼠模型中的肿瘤维持至关重要。在 N-MYC 扩增的 NB 细胞中 线，初步数据显示，使用小分子抑制剂靶向 WDR5 的 WIN 位点会导致 全基因组范围内 WDR5 结合减少，随后特定 N-MYC- 处 N-MYC 结合丧失 WDR5 共结合基因。重要的是，使用基因突变体破坏 N-MYC-WDR5 相互作用 损害 N-MYC 驱动锚定独立生长的能力，提供证据表明 与 N-MYC 相关的重要肿瘤功能需要与 WDR5 相互作用。这些数据共同提供了 N-MYC 需要 WDR5 来结合和调节 N-MYC-WDR5 共结合这一观点奠定了坚实的基础 对于驱动 N-MYC 特定活动很重要的基因。这个项目的目标是询问 N- N-MYC 扩增神经母细胞瘤细胞系中 MYC-WDR5 相互作用并确定 WDR5 利用遗传和基因在 N-MYC 驱动的转录过程和神经母细胞瘤功能上发挥作用 化学扰动。具体目标 1 将采用遗传和基因组方法来鉴定基因 与 N-MYC 和 WDR5 结合，并确定 N-MYC 的募集在哪些基因上 依赖于 N-MYC-WDR5 相互作用。具体目标 2 将结合高分辨率转录组学 通过细胞和体内功能测定进行分析，以挑战 N-MYC-WDR5 的重要性 相互作用。完成这些研究后，我们将确定 N-MYC 的精确基因 结合和转录受 N-MYC-WDR5 相互作用调节，并直接挑战 N-MYC-WDR5 相互作用对 NB 肿瘤发生多个方面的重要性。

项目成果

WD Repeat Domain 5 Inhibitors for Cancer Therapy: Not What You Think.

• DOI: 10.3390/jcm13010274
• 发表时间: 2024-01-03
• 期刊: JOURNAL OF CLINICAL MEDICINE
• 影响因子: 3.9
• 作者: Weissmiller, April M.;Fesik, Stephen W.;Tansey, William P.
• 通讯作者: Tansey, William P.