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.

项目概要 在美国，每年有近千名儿童被诊断患有神经母细胞瘤 (NBL)，并且 迄今为止，这种癌症是婴儿中最常见的类型。遗憾的是，高风险 NBL 约占 70% 所有 NBL 病例的五年生存率<50%。高风险 NBL 的特点是 MYCN 基因的扩增和第一染色体 1p36 (D1p36) 区域的缺失， 含有多种肿瘤抑制因子。但这些特征很难用作药理学用途 MYCN 的抑制已被证明是难以捉摸的，并且在之后重新激活肿瘤抑制基因的表达 基因组丢失几乎是不可能的。与 MYCN 在增加全球影响力方面的既定作用一致 转录，具有扩增的 MYCN 表达的高风险 NBL 独特地依赖于细胞 转录和RNA处理装置。对 MYCN 功能至关重要的机制是 Integrator Complex，负责共转录剪接所必需的 UsnRNA 生物合成 促进 RNA 聚合酶 II (RNAPII) 的周转和可塑性。积分器是一个 14 元复合体 与 RNAPII 相关，是调节转录输出的基础。我的实验室已经 研究Integrator的分子机制十多年（19-34），我们发现： 积分子亚基 11 (INTS11) 是复合物的关键酶成分。 INTS11 利用其 RNA 核酸内切酶活性可切割剪接体 UsnRNA 的 3' 端，因此是预剪接所需的 mRNA 剪接。此外，我们最近发现 INTS11 对于裂解新生蛋白也是必需的—— 编码RNA以促进RNAPII循环。这两个功能对于支持高转录至关重要 当 MYCN 过量产生时观察到。除了 MYCN 扩增肿瘤所需之外，最 提名 Integrator 作为 NBL 中潜在药物靶标的一个有吸引力的特征是 INTS11 基因是 位于 1p36 内，在 NBL 中经常被删除。为了支持这一观察，DEPMAP 的数据 揭示 NBL 细胞系对 INTS11 的部分耗尽高度敏感，这表明 NBL 细胞系具有独特且 对该基因的强烈依赖性。总而言之，这些观察结果支持这样一个假设：INTS11 代表了 NBL 中一个新颖且未经探索的目标（图 1），我们下面的具体目标旨在 直接测试这个。具体目标 1. 确定 MYCN 扩增和 D1p36 NBL 细胞系的敏感性 INTS11 下调。具体目标 2. 测试 INTS11 候选抑制剂的不同毒性 高风险 NBL 细胞系。