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，每年有近千名儿童被诊断患有神经母细胞瘤（NBL）， 到目前为止，这种癌症是婴儿中最常见的类型。可悲的是，高风险的NBL，其中包括约70%的 所有NBL病例的五年生存率<50%。高风险NBL的特点是具有 MYCN基因扩增和1号染色体1p36（D1p36）区域缺失， 有多种肿瘤抑制剂但是这些特征很难作为药理学来利用 MYCN的抑制已被证明是难以捉摸的，并且在MYCN抑制后重新激活肿瘤抑制基因表达， 基因组丢失几乎是不可能的。与MYCN在增加全球 转录，具有扩增MYCN表达的高风险NBL独特地依赖于细胞内 转录和RNA加工装置。对MYCN功能至关重要的机制之一是 整合子复合物，其负责共转录剪接所必需的UsnRNA生物合成 并促进RNA聚合酶II（RNAPII）的周转和可塑性。Integrator是一个14元复合物 与RNAPII相关，并且是调节转录输出的基础。我的实验室 研究Integrator的分子机制超过十年（19 - 34），我们发现， 整合子亚基11（INTS 11）是复合物的关键酶组分。INTS11利用其 RNA内切酶活性切割剪接体UsnRNA的3'-末端，因此是前 mRNA剪接。此外，我们最近发现INTS 11也是切割新生蛋白质所必需的- 编码RNA以促进RNAPII再循环。这两种功能对于支持高转录都至关重要 当MYCN过度生产时观察到。除了在MYCN扩增的肿瘤中需要外， 将整合子提名为NBL中潜在的可药物靶点的吸引人的特征是，INTS 11基因是 位于1p36内，并且在NBL中经常缺失。为支持这一观察，环境监测和评估方案提供的数据 揭示了NBL细胞系对INTS 11的部分缺失高度敏感，这表明了一种独特的， 对这个基因的强烈依赖。总之，这些观察结果支持一个假设，即INTS 11 代表了NBL中一个新的和未探索的目标（图1），我们下面的具体目标旨在 直接测试这个。具体目标1。测定MYCN扩增和D1p36 NBL细胞系的敏感性 INTS11下调。具体目标2。测试INTS 11的候选抑制剂在小鼠中的差异毒性。 高风险NBL细胞系。