Credentialing Delta-like 3 (DLL3) as an oncoprotein and immunotherapeutic target in neuroblastoma

证明 Delta-like 3 (DLL3) 作为神经母细胞瘤的癌蛋白和免疫治疗靶点

• 批准号: 9907428
• 负责人: Nathan Michael Kendsersky
• 金额: $ 4.55万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9907428
• 关键词: 3-Dimensional; ASCL1 gene; Adult; Antibody Therapy; Bioinformatics; Biological; Biological Assay; Biology; CRISPR/Cas technology; Cancer Burden; Cell Line; Cell Proliferation; Cell Surface Proteins; Cell Survival; Cell surface; Cells; Child; Childhood Solid Neoplasm; Clinical Research; Clinical Trials; Clustered Regularly Interspaced Short Palindromic Repeats; Credentialing; Data; Dependence; Development; Down-Regulation; ES01; Enhancers; Environment; GATA3 gene; GTP-Binding Protein alpha Subunits, Gs; Gene Silencing; Genes; Genetic; Genetic Enhancer Element; Genetic Transcription; Genomics; Glycosphingolipids; Immune; Immunotherapeutic agent; Immunotherapy; Invaded; Knock-out; Lead; Ligands; MYCN gene; Malignant Childhood Neoplasm; Malignant Neoplasms; Mediating; Membrane Proteins; Methodology; Mission; Modeling; Molecular Biology; National Research Service Awards; Nature; Neural Cell Adhesion Molecule L1; Neural Crest; Neuroblastoma; Neuroendocrine Tumors; Oncogenes; Oncogenic; Oncoproteins; Outcome; Output; Pain; Pathway interactions; Patients; Peripheral Nervous System; Pharmacology; Phenotype; Proteins; Proteomics; Public Health; Regulation; Relapse; Research; Role; Scientist; Signal Pathway; Signal Transduction; Surface; Survival Rate; Survivors; Sympathetic Nervous System; Testing; Therapeutic; Undifferentiated; United States National Institutes of Health; Work; Xenograft procedure; base; cancer diagnosis; cell type; chimeric antigen receptor T cells; clinically actionable; comorbidity; differential expression; disorder risk; epigenomics; evidence base; gamma secretase; genomic aberrations; high risk; in vivo; infancy; inhibitor/antagonist; insight; knock-down; lung small cell carcinoma; malignant state; multimodality; neoplastic cell; neuroblastoma cell; notch protein; novel; novel therapeutics; overexpression; preclinical development; preclinical study; pressure; side effect; small hairpin RNA; targeted treatment; therapeutic target; transcription factor; transcriptome sequencing; tumor; tumorigenesis

Project Summary Neuroblastoma (NBL) is a pediatric solid tumor that arises from deregulated development of the sympathetic nervous system. The five-year survival of patients with high-risk NBL is only 40% and relapse-free survival is extremely rare. This underscores the need to identify biologically-relevant and clinically-actionable targets for children with high-risk disease. One genomic aberration found in high-risk NBL is amplification of a lineage- specific oncogene, MYCN. Once amplified, the MYCN transcription factor can invade enhancers of other lineage oncogenes and establish a core regulatory circuit (CRC), whereby these transcription factors autoregulate themselves and each other to maintain expression of the circuit. Due to the developmental nature of NBL, these tumors display unique cell-surface molecules and may be susceptible to immunotherapy. An antibody-based therapy (dinutuximab) targeting the glycosphingolipid molecule, GD2, extends the 5-year survival rates of patients with aggressive, high-risk NBL. However, GD2 is also found on cells of the peripheral nervous system and treatment with dinutuximab causes agonizing pain. To avoid these on-target / off-tumor side effects, we aim to identify NBL tumor-specific surface molecules that are regulated by the CRC and associated with the undifferentiated, malignant state of NBL. Our preliminary analysis of CRC-bound genes revealed both novel and known immunotherapeutic targets, including ALK and L1CAM. Another CRC-regulated cell-surface protein identified in our analysis is Delta-like canonical notch ligand 3 (DLL3). DLL3 has NBL-specific cell-surface expression, and is known as an inhibitor of Notch signaling in other cell types. Clinical trials with DLL3-targeted immunotherapies in adult cancers are currently ongoing, however, the oncogenic mechanism of DLL3 in these tumor-types, and in NBL, is uncharacterized and poorly understood. In this proposal, we also aim to uncover the relevant biology of DLL3 in NBL in order to credential DLL3 as a viable immunotherapeutic target for this pediatric malignancy. Our preliminary data shows that shRNA-depletion of DLL3 reduces the cellular viability in NBL cell line models with high expression of DLL3. Furthermore, CRISPR-Cas9 genomic depletion of DLL3 results in increased expression of a canonical Notch target gene, HES1. Therefore, we postulate that DLL3 inhibits Notch signaling to promote persistent NBL cell survival, and that the CRC transcriptionally regulates DLL3 to cause overexpression. This project will combine integrative approaches in bioinformatics and molecular biology to 1) describe differentially expressed, cell-surface targets driven by the CRC and 2) determine the role of DLL3 in the malignant state of NBL. We also anticipate that we can apply our CRC computational workflow to other cancers with established transcriptional circuits. This NRSA F31 will provide insight into NBL biology and the regulation of candidate immunotherapeutic targets in NBL, both of which will inform preclinical and clinical studies with targeted immunotherapies.

项目摘要 神经母细胞瘤（NBL）是一种儿科实体肿瘤，由交感神经的不受管制的发展产生 神经系统。高危NBL患者的五年生存率仅为40％，无复发生存率为 极为罕见。这强调了确定与生物学上与生物学相关的临床和临床可靠性目标的需求 患有高风险疾病的儿童。在高风险NBL中发现的一种基因组像差是扩增谱系 特定的癌基因，mycn。一旦扩增，MYCN转录因子可以侵入其他谱系的增强子 癌基因并建立核心调节回路（CRC），这些转录因子自动调节 自己和彼此以保持电路的表达。由于NBL的发展性质，这些 肿瘤显示独特的细胞表面分子，可能容易受到免疫疗法的影响。基于抗体 靶向糖脂分子GD2的治疗（Dinutuximab）延长了5年生存率 具有侵略性，高风险NBL的患者。但是，在周围神经系统的细胞上也发现了GD2 Dinutuximab的治疗会导致痛苦。为了避免这些目标 /非肿瘤副作用，我们的目标 确定由CRC调节并与之相关的NBL肿瘤特异性表面分子 NBL的未分化，恶性状态。我们对CRC结合基因的初步分析揭示了新颖和 已知的免疫治疗靶标，包括ALK和L1CAM。另一个由CRC调节的细胞表面蛋白 在我们的分析中确定的是三角形的规范凹配体3（DLL3）。 DLL3具有NBL特异性的细胞表面 表达，被称为其他细胞类型中Notch信号传导的抑制剂。 DLL3目标的临床试验 然而，成年癌症的免疫疗法目前正在进行中 肿瘤类型，在NBL中是未经表征且了解不足的。在此提案中，我们还旨在发现 DLL3在NBL中的相关生物学，以使DLL3作为该儿科的可行免疫治疗靶标 恶性。我们的初步数据表明，dll3的shRNA耗尽降低了NBL细胞中的细胞活力 具有高表达DLL3的线模型。此外，DLL3的CRISPR-CAS9基因组耗竭导致 增加规范档位靶基因的表达，Hes1。因此，我们假设DLL3抑制了Notch 信号传导以促进持续的NBL细胞存活，CRC在转录中调节DLL3引起 过表达。该项目将将生物信息学和分子生物学中的综合方法结合到1） 描述由CRC驱动的差异表达的细胞表面目标，2）确定DLL3在 NBL的恶性状态。我们还预计我们可以将CRC计算工作流程应用于其他 具有既定转录电路的癌症。这种NRSA F31将提供有关NBL生物学和 NBL中候选免疫治疗靶标的调节，这两个都将为临床前和临床研究提供信息 具有靶向免疫疗法。