Targeted therapy against TERT oncogene-rearranged neuroblastoma

TERT癌基因重排神经母细胞瘤的靶向治疗

基本信息

批准号：
10452641
负责人：
Tao Liu
金额：
$ 5.4万
依托单位：
UNIVERSITY OF NEW SOUTH WALES
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10452641
关键词：
Acute Myelocytic Leukemia Apoptosis Cancer Patient Cell Line Cell Proliferation Cell Survival Childhood Malignant Brain Tumor Clinical Trials Data Data Set Enhancers Gene Expression Genes Genetic Transcription Human In Vitro Life MDM2 gene Mus Neuroblastoma Normal tissue morphology Oncogenes Outcome Patients Phosphorylation Phosphotransferases Play RNA Polymerase II Refractory Relapse Role Serine Solid Neoplasm TERT gene TP53 gene Telomerase Inhibitor Testing Tissues Toxic effect Transcription Elongation Transcription Initiation Tumor Tissue Xenograft procedure anti-cancer cell growth childhood cancer mortality early childhood high risk in vivo inhibitor knock-down leukemia mouse model neuroblastoma cell new therapeutic target overexpression patient prognosis promoter protein degradation protein expression side effect small hairpin RNA targeted treatment transcriptome sequencing tumor tumor progression tumorigenesis

项目摘要

PROJECT SUMMARY/ABSTRACT Background. TERT oncogene rearrangement with transcriptional super-enhancers leads to substantial TERT over-expression and neuroblastoma in high-risk neuroblastoma patients. Telomerase inhibitors show little anticancer effects and cause life-threatening side effects in childhood brain cancer patients in clinical trials. The transcriptional kinases CDK7 and CDK9 play critical roles in super-enhancer-associated oncogene transcriptional initiation, pause release and elongation respectively, and CKIα induce p53 protein degradation. The CDK7/CDK9/CKIα co-inhibitor A51 suppresses super-enhancer-associated oncogene expression and activates p53 protein expression, resulting in leukemia regression in mice with no toxicity to normal tissues. Preliminary Data. We have found that CDK7/CDK9/CKIα co-knockdown with shRNAs or treatment with the CDK7/CDK9/CKIα co-inhibitor A51 considerably reduced TERT gene expression and MDM2 protein expression, activated p53 protein expression, and induced substantial apoptosis in TERT oncogene- rearranged neuroblastoma, but not normal cells. Analysis of a RNA sequencing gene expression-patient prognosis dataset from 493 neuroblastoma patients, showed that high levels of CDK7 gene expression in human tumor tissues positively correlated with high levels of TERT gene expression, and that high levels of CDK7 and CKIα expression in human neuroblastoma tissues correlated with poor patient prognosis. Specific Aims. (1) To identify the critical roles of CDK7 and CDK9 in inducing TERT gene transcriptional initiation, elongation and over-expression and CKIα in inducing p53 protein degradation in TERT-rearranged neuroblastoma cells; (2) To identify the critical roles of CDK7, CDK9 and CKIα in TERT-rearranged neuroblastoma cell proliferation and survival in vitro and tumor progression in vivo; (3) To define the anticancer efficacy of the CDK7/CDK9/CKIα co-inhibitor A51 against TERT-rearranged neuroblastoma in vitro and in vivo. Outcomes and Significance. We hope to demonstrate that CDK7 and CDK9 co-operatively induce TERT gene transcriptional initiation, pause release, elongation and over-expression, and CKIα induces p53 protein degradation; that CDK7, CDK9 and CKIα co-operatively induce TERT-rearranged neuroblastoma cell proliferation and survival in vitro and tumor progression in vivo; and that the CDK7/CDK9/CKIα co-inhibitor A51 efficiently blocks TERT gene transcriptional initiation, pause release, elongation and expression, activates p53 protein expression, induces neuroblastoma cell apoptosis in vitro, and blocks tumor progression and causes tumor regression in mice xenografted with TERT-rearranged neuroblastoma cell lines or patient-derived neuroblastoma tissues. As A51 is currently in clinical trials in leukemia patients, completion of this project will provide the vital evidence for clinical trials of A51 therapy in patients with TERT gene-rearranged neuroblastoma, against which no targeted therapy is now available for, or has ever be tested in, clinical trials.

项目概要/摘要背景：转录超级增强子的 TERT 癌基因重排导致大量的 TERT。端粒酶抑制剂在高危神经母细胞瘤患者中的过度表达与神经母细胞瘤关系不大。在临床试验中，它对儿童脑癌患者具有抗癌作用并导致危及生命的副作用。转录激酶 CDK7 和 CDK9 在超级增强子相关癌基因中发挥关键作用转录起始、暂停释放和延伸，CKIα 诱导 p53 蛋白降解。 CDK7/CDK9/CKIα 共抑制剂 A51 抑制超级增强子相关癌基因的表达并激活 p53 蛋白表达，导致小鼠白血病消退，且对正常组织无毒性。初步数据我们发现 CDK7/CDK9/CKIα 与 shRNA 共同敲低或用 CDK7/CDK9/CKIα 共抑制剂 A51 显着降低 TERT 基因表达和 MDM2 蛋白表达，激活 p53 蛋白表达，并诱导 TERT 癌基因大量凋亡重排神经母细胞瘤，但不重排正常细胞的 RNA 测序基因表达分析。来自 493 名神经母细胞瘤患者的预后数据集显示，CDK7 基因高水平表达人类肿瘤组织与高水平的TERT基因表达呈正相关，并且高水平的TERT基因表达与高水平的TERT基因表达呈正相关。人神经母细胞瘤组织中 CDK7 和 CKIα 的表达与患者不良预后相关。具体目的 (1) 确定CDK7和CDK9在诱导TERT基因转录中的关键作用。 TERT重排中起始、延伸和过度表达以及CKIα诱导p53蛋白降解 (2) 确定CDK7、CDK9和CKIα在TERT重排中的关键作用； (3) 抗癌的定义 CDK7/CDK9/CKIα 共抑制剂 A51 在体外和体内对 TERT 重排神经母细胞瘤的功效。结果和意义我们希望证明 CDK7 和 CDK9 协同诱导 TERT。基因转录起始、暂停释放、延伸和过度表达，以及 CKIα 诱导 p53 蛋白 CDK7、CDK9 和 CKIα 协同诱导 TERT 重排的神经母细胞瘤细胞体外增殖和存活以及体内肿瘤进展；以及 CDK7/CDK9/CKIα 联合抑制剂 A51 有效阻断 TERT 基因转录起始、暂停释放、延伸和表达，激活 p53 蛋白表达，在体外诱导神经母细胞瘤细胞凋亡，并阻止肿瘤进展和原因 TERT重排神经母细胞瘤细胞系或患者来源的异种移植小鼠的肿瘤消退由于 A51 目前正在白血病患者中进行临床试验，该项目的完成将为A51治疗TERT基因重排患者的临床试验提供重要证据神经母细胞瘤，目前还没有针对这种肿瘤的靶向治疗，也没有在临床试验中进行过测试。