Targeting Transcriptional Elongation in Pediatric Glioma

靶向小儿胶质瘤的转录延伸

基本信息

批准号：
10829524
负责人：
Rintaro Hashizume
金额：
$ 37.81万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-01 至 2027-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10829524
关键词：
Acetylation Animals Astrocytes Biological Brain CRISPR/Cas technology Cancer Patient Cells ChIP-seq Characteristics Child Childhood Brain Neoplasm Childhood Glioma Chromatin Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats Combined Modality Therapy Complex DNA Damage DNA Repair DNA Repair Pathway Data Dependence Development Diagnosis Diffuse intrinsic pontine glioma Disease Drug Kinetics Elongation Factor Epigenetic Process Expression Profiling Foundations Gene Expression Gene Mutation Genes Genetic Genetic Screening Genetic Transcription Goals Growth Histone H3 Histones Human In Vitro Individual Knock-out Knowledge Lysine Malignant Childhood Neoplasm Mediating Methionine Methylation Mission Modeling Modification Molecular Mutation National Institute of Neurological Disorders and Stroke Nature Nervous System Oncogenic Pathogenesis Patient-Focused Outcomes Patients Polymerase Positive Transcriptional Elongation Factor B Property Public Health RNA RNA Polymerase II Radiation Radiation Tolerance Radiation enhancer Radiation therapy Rare Diseases Research Resistance Sequence Analysis Tertiary Protein Structure Testing Therapeutic Therapeutic Intervention Toxic effect Transcription Elongation Transcription Initiation Transcriptional Activation Treatment outcome Xenograft procedure bioluminescence imaging chemotherapeutic agent clinical practice combinatorial early phase clinical trial effective therapy experimental study genome-wide improved in vivo inhibitor inhibitor therapy knock-down mutant neoplastic cell nervous system disorder new therapeutic target novel novel therapeutic intervention patient derived xenograft model patient screening pharmacologic prevent protein H(3)research clinical testing small hairpin RNA small molecule small molecule inhibitor targeted treatment therapeutic target therapeutically effective therapy resistant transcriptional reprogramming transcriptome transcriptome sequencing tumor tumor growth tumor progression

项目摘要

PROJECT SUMMARY/ABSTRACT - Diffuse intrinsic pontine glioma (DIPG) is one of the most devastating pediatric cancers. Numerous clinical trials in decades, involving different combinations of chemotherapeutic agents and radiation, have been ineffective in treating DIPG. The identification of efficacious therapeutic targets based on the molecular characteristic is of high importance for improving treatment outcomes for children with DIPG. The discovery of oncogenic histone gene mutations in DIPG has dramatically improved our understanding of disease pathogenesis, and stimulated the development of novel therapeutic approaches to target epigenetic modifiers. We have recently shown that targeted bromo- and extra-terminal (BET) domain protein 4 (BRD4) activity using JQ1 inhibitor results in a significant delay of tumor progression and prolonged survival of animals bearing DIPG patient-derived xenograft (PDX). Because of their promising anti-tumor activity, BRD4 inhibitors are being tested in a number of cancer patient clinical trials. However, tumors that initially respond to small molecule inhibitor therapies, such as those targeting BRD4 activity, eventually become resistant to monotherapy treatment, affirming the need for more effective therapeutic interventions. In order to identify new effective therapeutic targets, and discover novel combinatorial approaches to prevent or delay acquired resistance to monotherapy, we performed an unbiased genome-wide CRISPR/Cas9-based genetic screen of patient-derived DIPG cells. We identified nine “network modules” that are significantly enriched in CRISPR targets. One of these modules includes POLR2I, which encodes a subunit of RNA polymerase II (Pol II) that is involved in transcription elongation. We subsequently observed that targeting POLR2I activity through short-hairpin RNA knockdown and treatment of the small-molecule Pol II inhibitors block transcriptional elongation and inhibit the growth of DIPG in vitro and in vivo. Here, we will test the hypothesis that inhibition of Pol II transcriptional elongation in combination with BRD4 inhibition will further suppress gene transcription and will either delay or prevent DIPG from acquiring resistance to monotherapy. This dual inhibition approach will interfere with gene transcription at two levels: transcriptional initiation (BRD4) and elongation (Pol II). This project will also explore how these targeted therapies interact with radiation in treating DIPG, which is important due to the use of radiation in treating nearly all cases of DIPG in children. The successful completion of proposal study has significant impact on clinical practice and accumulating data from this research could therefore lay the foundation for early clinical trials of this approach, given the high unmet need and orphan disease status of DIPG.

项目摘要/摘要 - 弥漫性固有的庞然神经胶质瘤（DIPG）是最具破坏性的小儿癌。几十年来，许多临床试验涉及化学治疗的不同组合药物和辐射在治疗DIPG方面无效。识别有效的治疗靶标基于分子特征，对于改善儿童的治疗结果至关重要 dipg。 DIPG中致癌组蛋白基因突变的发现极大地改善了我们的理解疾病发病机理，并刺激了靶向遗传学的新型治疗方法的发展修饰符。我们最近表明，针对的溴和末端（BET）结构域蛋白4（BRD4）使用JQ1抑制剂的活性导致肿瘤进展的显着延迟和动物的长期存活率轴承DIPG患者衍生的Xenographic（PDX）。由于其承诺的抗肿瘤活性，BRD4抑制剂正在许多癌症患者临床试验中进行测试。但是，最初对小的肿瘤分子抑制剂疗法，例如靶向BRD4活性的分子疗法，有时会抗药性治疗，影响更有效的治疗干预措施的需求。为了确定新的有效治疗靶标，并发现新颖的组合方法，以防止或延迟获得的抵抗力单一疗法，我们进行了公正的全基因组CRISPR/CAS9的基于患者衍生的遗传筛查 DIPG细胞。我们确定了9个“网络模块”，这些模块大大富含CRISPR目标。其中之一模块包括POLR2I，它编码参与转录的RNA聚合酶II（POLII）的亚基伸长。随后，我们观察到，通过短发的RNA敲低靶向polr2i活性和小分子POL II抑制剂的治疗阻断转录伸长并抑制DIPG的生长体外和体内。在这里，我们将检验以下假设：与BRD4抑制结合将进一步抑制基因转录，并将延迟或防止DIPG 从获得耐药性到单一疗法。这种双重抑制方法将干扰基因转录两个级别：转录倡议（BRD4）和伸长率（POL II）。该项目还将探索如何靶向疗法在治疗DIPG中与辐射相互作用，这很重要，因为使用辐射治疗几乎所有儿童dipg案件。提案研究的成功完成对因此，这项研究的临床实践和积累数据可以为早期临床奠定基础鉴于DIPG的高未满足需求和孤儿疾病状况，这种方法的试验。