Probing INTS11 as a novel target in neuroblastoma

探索 INTS11 作为神经母细胞瘤的新靶点

• 批准号: 10577214
• 负责人: ERIC J WAGNER
• 金额: $ 7.7万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-07 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10577214
• 关键词: 1p36; Anabolism; Biological; Cell Line; Cell Proliferation; Cells; Child; Chromosomes; Code; Complex; Coupled; Data; Dependence; Diagnosis; Down-Regulation; Endoribonucleases; FDA approved; Gene Expression; Genes; Genetic Transcription; Genomics; Infant; Laboratories; MYCN gene; Malignant Neoplasms; Measures; Molecular; Mutation; Neuroblastoma; Normal Cell; Output; Pharmaceutical Preparations; Property; Proteins; RNA; RNA Polymerase II; RNA Processing; RNA Splicing; Reagent; Recycling; Reporter; Research; Research Project Grants; Role; Running; Spliceosomes; Survival Rate; System; Testing; Toxic effect; Tumor Suppressor Genes; Tumor Suppressor Proteins; Tumorigenicity; United States; design; druggable target; high risk; high throughput screening; inhibitor; knock-down; mRNA Precursor; member; neuroblastoma cell; new therapeutic target; novel; patient prognosis; pharmacologic; small hairpin RNA; small molecule; small molecule inhibitor; tumor; virtual

PROJECT SUMMARY Nearly a thousand children are diagnosed each year with Neuroblastoma (NBL) in the United States, and this cancer is, by far, the most prevalent type in infants. Sadly, high-risk NBL, which comprises ~70% of all NBL cases, has a five-year survival rate of <50%. High-risk NBL is characterized by having amplification of the MYCN gene and deletion of the 1p36 (D1p36) region of chromosome one, which houses multiple tumor suppressors. But these signatures are difficult to exploit as pharmacologic inhibition of MYCN has proven elusive, and re-activating tumor suppressor gene expression after genomic loss is virtually impossible. Consistent with MYCN’s established role in increasing global transcription, high-risk NBLs with amplified MYCN expression are uniquely dependent on the cellular transcription and RNA processing apparatus. Among this machinery vital to MYCN function is the Integrator Complex, which is responsible for UsnRNA biosynthesis essential for co-transcriptional splicing and promoting RNA polymerase II (RNAPII) turnover and plasticity. Integrator is a 14-membered complex associated with RNAPII and is fundamental in regulating transcriptional output. My laboratory has been investigating the molecular mechanism of Integrator for over ten years(19-34), and we have found that Integrator subunit 11 (INTS11) is the critical enzymatic component of the complex. INTS11 utilizes its RNA endonuclease activity to cleave the 3'-end of spliceosomal UsnRNA and is thus required for pre- mRNA splicing. Moreover, we recently found that INTS11 is also necessary to cleave nascent protein- coding RNA to promote RNAPII recycling. Both functions are critical to support high transcription observed when MYCN is overproduced. Beyond being required in MYCN-amplified tumors, the most attractive feature nominating Integrator as a potential druggable target in NBL is that the INTS11 gene is located within 1p36 and is frequently deleted in NBLs. In support of this observation, data from DEPMAP reveals that NBL cell lines are highly sensitive to partial depletion of INTS11, indicating a unique and robust dependency on this gene. Altogether, these observations support a hypothesis that INTS11 represents a novel and unexplored target in NBL (Fig. 1), and our Specific Aims below are designed to test this directly. Specific Aim 1. Determine the sensitivity of MYCN-amplified and D1p36 NBL cell lines to INTS11 downregulation. Specific Aim 2. Test candidate inhibitors of INTS11 for differential toxicity in high-risk NBL cell lines.

项目摘要 在美国，每年将近一千名儿童患有神经母细胞瘤（NBL）， 到目前为止，这种癌症是婴儿中最普遍的类型。可悲的是，高风险NBL，占约70％ 所有NBL病例的生存率<50％。高风险NBL的特征是 MYCN基因的扩增和1p36（D1p36）区域的染色体的删除 容纳多种肿瘤补充剂。但是这些签名很难用作药学 抑制MYCN已证明难以捉摸，并且在 基因组损失几乎是不可能的。与MYCN在增加全球方面的既定作用一致 转录，具有扩增MYCN表达的高风险NBL独特取决于细胞 转录和RNA处理设备。在此对MYCN功能至关重要的机械中是 集成仪复合物，负责USNRNA生物合成对于共同剪接必不可少的 并促进RNA聚合酶II（RNAPII）周转率和可塑性。集成商是一个14元的复合物 与RNAPII相关，并且在控制转录输出方面至关重要。我的实验室已经 研究整合物的分子机制已有十多年（19-34），我们发现 集成子亚基11（INTS11）是复合物的关键酶成分。 INTS11利用它 RNA核酸内切酶的活性清除剪接体USNRNA的3'末端，因此需要 mRNA剪接。此外，我们最近发现，清除新生蛋白的INTS11也是必要的 编码RNA以促进RNAPII回收。这两个功能对于支持高转录至关重要 观察到MYCN生产过量时。除了在MyCN放大肿瘤中需要 吸引人的特征提名集成剂是NBL中潜在的可药物靶标的，即Ints11基因是 位于1P36内，经常以NBL删除。为了支持此观察，来自DEPMAP的数据 揭示了NBL细胞系对INTS11的部分耗竭高度敏感，这表明是独特的和 对此基因的强大依赖。总之，这些观察结果支持一个假设，即INTS111111 代表NBL中的一个新颖而意外的目标（图1），我们以下具体目的旨在 直接测试。具体目标1。确定MYCN放大和D1P36 NBL细胞系的灵敏度 到INTS11下调。特定目的2。ints11的测试候选抑制剂对差异毒性 高风险的NBL细胞系。