Dissecting the contribution of the transcriptional regulators of SNS fate to neuroblastoma oncogenesis

剖析 SNS 命运转录调节因子对神经母细胞瘤肿瘤发生的贡献

• 批准号: 10179331
• 负责人: KEVIN W FREEMAN
• 金额: $ 34.77万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-06-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10179331
• 关键词: 1p36; Address; Agar; Binding; Bromodomain; Cancer Patient; Cancer cell line; Cell Line; Cells; Cessation of life; Child; Combined Modality Therapy; Data; Death Rate; Development; Diagnosis; Disease; Disease Resistance; Engineering; Enhancers; Epigenetic Process; Genetic; Genetic Transcription; Goals; Grant; Growth; Histones; Human; Impairment; Individual; Loss of Heterozygosity; Maintenance; Malignant - descriptor; Malignant Childhood Neoplasm; Malignant Neoplasms; Modeling; Mus; Mutate; Mutation; Neural Crest; Neuroblastoma; Neurons; Oncogenes; Pathway interactions; Patients; Pharmaceutical Preparations; Primary Neoplasm; Property; Proteins; Reader; Recurrent disease; Relapse; Resistance; Role; Standard Model; Sympathetic Nervous System; System; Tertiary Protein Structure; Testing; Therapeutic Effect; Time; Tumor Cell Line; Work; base; behavior in vitro; blocking factor; high risk; improved; infancy; inhibitor/antagonist; nervous system development; neuroblastoma cell; new therapeutic target; novel; novel therapeutic intervention; novel therapeutics; overexpression; patient derived xenograft model; programs; promoter; relapse patients; response; standard of care; stem cell differentiation; stem cells; therapeutic target; transcription factor; tumor; tumor xenograft; tumorigenesis; tumorigenic

Project Summary/Abstract: Neuroblastoma (NB) is the most common cancer of infancy and arises from immature cells of the sympathetic nervous system (SNS) that derive from neural crest stem cells (NCCs). NB is responsible for 15% of pediatric cancer deaths with the majority of high-risk NB patients eventually relapsing with resistant disease. Finding novel treatments, especially for relapse disease, is desperately needed for high-risk neuroblastoma. The standard model of neuroblastoma initiation assumes a block in differentiation. We propose that SNS factors important for the growth and establishment of the SNS are co-opted by neuroblastoma mutations to instead support the growth and establishment of the tumor. Further based on our preliminary findings we propose a class of drugs called BET inhibitors, which were found to be very effective against neuroblastoma, partially function by suppressing important SNS factors. Here, we hypothesize that SNS cell fate regulators, are critical for NB oncogenesis and responsible for NB sensitivity to BET inhibitors. The two aims we will pursue in this proposal: Aim 1: Are regulators of cell fate critical targets of BET inhibition in NB? Aim 2: Does cell identity/developmental stage contribute to malignant transformation of primary NCCs? For the first aim we will determine in neuroblastoma cell lines, if BET inhibitors work through turning off critical SNS factors. Thus, here we will: 1) Interrogate the impact of BET inhibitors on SNS lineage factors expression. 2) Investigate loss of the BET protein BRD4 binding in response to the BET inhibitor JQ1, in human NB cell lines. 3) Test if silencing SNS lineage factors individually or in combination interferes with growth of human NB cell lines and NB tumors. 4) Determine if lineage factors expressed in trans from the JQ1-insensitive MSCV promoter conveys resistance to BET inhibitors in human NB cell lines and patient derived xenograft tumors. Aim 1 will establish if SNS lineage factors are necessary for NB maintenance. In Aim 2, we will interrogate if these factors are functionally required for NB oncogenesis. To test the second aim we will use a unique system we have developed for turning normal immature SNS mouse cells, the NCCs, into neuroblastoma to determine if the important SNS factors can contribute to initiating neuroblastoma. For the second aim we will: 1) Determine if the SNS core factors block NCC differentiation into sympathetic neurons. 2) We will determine if the SNS core transcription factors promote tumorigenic behavior in vitro. 3) Determine if the SNS core transcription factors promotes the formation of NCC-derived NB tumors alone or in combination with N-Myc or 1p36 loss of heterozygosity, two common mutations in NB. Overall, our work will inform us of whether factors important for SNS development also contribute to neuroblastoma. Further our ability to engineer neuroblastoma tumors from immature SNS mouse cells uniquely enables us to illuminate novel mechanisms of neuroblastoma sensitivity to BET inhibition and discover new genetic contributors to this disease.

项目摘要/摘要： 神经母细胞瘤（NB）是婴儿期最常见的癌症，来自交感神经的未成熟细胞 神经系统（SNS）源自神经rest干细胞（NCC）。 NB负责15％的小儿 大多数高危NB患者的癌症死亡最终会因抗性疾病而复发。发现 高风险神经母细胞瘤迫切需要新的治疗，尤其是用于复发疾病。这 神经母细胞瘤启动的标准模型假定分化的障碍。我们建议SNS因素 对于神经母细胞瘤突变，对SN的生长和建立很重要 支持肿瘤的生长和建立。进一步基于我们的初步发现，我们提出了 称为BET抑制剂的药物类别，发现对神经母细胞瘤非常有效，部分地 通过抑制重要的SNS因子来起作用。在这里，我们假设SNS细胞命运调节剂是 对于NB的肿瘤发生至关重要，并使NB对BET抑制剂的敏感性至关重要。我们两个目标 在此提案中会追求：目标1：细胞命运的监管因子是否在NB中抑制BET的关键目标？目标2： 细胞身份/发育阶段是否有助于原发性NCC的恶性转化？第一个 目的，我们将在神经母细胞瘤细胞系中确定，如果BET抑制剂通过关闭关键SNS起作用 因素。因此，我们将在这里：1）询问BET抑制剂对SNS谱系因子表达的影响。 2） 在人NB细胞系中，研究响应BET抑制剂JQ1的BET蛋白BRD4结合的损失。 3）测试是否单独或组合干扰了人NB细胞的生长，是否测试SNS谱系因子 线和NB肿瘤。 4）确定在从JQ1不敏感的MSCV的反式中表达的谱系因子是否表达 启动子在人NB细胞系和患者衍生的异种移植肿瘤中传达了对BET抑制剂的抗性。 AIM 1将确定NB维护是否需要SNS谱系因素。在AIM 2中，我们会询问 这些因素在功能上是NB肿瘤发生的。为了测试第二个目标，我们将使用唯一的系统 我们已经开发用于将正常未成熟的SNS小鼠细胞（NCC）转变为神经母细胞瘤 如果重要的SNS因子可以有助于启动神经母细胞瘤。对于第二个目标，我们将：1） 确定SNS核心因素是否阻止NCC分化为交感神经元。 2）我们将确定是否 SNS核心转录因子在体外促进了肿瘤性行为。 3）确定SNS核心是否 转录因子促进单独或与N-MYC结合的NCC衍生的NB肿瘤的形成 或1P36杂合性丧失，NB中有两个共同的突变。总体而言，我们的工作将告知我们是否 SNS发育重要的因素也有助于神经母细胞瘤。进一步我们的设计能力 来自未成熟SNS小鼠细胞的神经母细胞瘤肿瘤独特地使我们能够照亮新的机制 神经母细胞瘤对抑制的敏感性并发现对这种疾病的新遗传因素。