Examining the Role of a Novel Long Noncoding RNA, linc02454, in Resistance of Glioblastoma to Temozolomide

检查新型长非编码 RNA (linc02454) 在胶质母细胞瘤对替莫唑胺耐药中的作用

基本信息

批准号：
10487542
负责人：
Frank Attenello
金额：
$ 19.26万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
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：
2021-09-15 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10487542
关键词：
12q14 ATAC-seq Adjuvant Therapy Affect Binding Biological Assay Biology Bioluminescence Brain Neoplasms CXC Chemokines Cell Culture Techniques Cell Cycle Cell Line Cell Survival Cells Chemoresistance Chemotherapy and/or radiation Chromatin Chromosomes Clinical Trials CpG Island Methylator Phenotype DNA Data Data Set Dependence Development Plans Diagnostic Down-Regulation Epigenetic Process Excision Exposure to Feasibility Studies Fluorescent in Situ Hybridization Funding Gene Amplification Gene Expression Genes Genetic Genetic Epistasis Genetic Transcription Genome Glioblastoma Glioma Goals In Vitro International Malignant Neoplasms Mass Spectrum Analysis Mentors Mentorship Methylation Mus Neurosurgeon Operative Surgical Procedures Patients Phenotype Primary Brain Neoplasms Proteins RNA RNA methylation Radiation therapy Recurrence Recurrent tumor Regimen Regulation Repression Research Training Resistance Role Sampling Scientist Supervision Surgeon Techniques Testing The Cancer Genome Atlas Training Transcript Transcriptional Regulation Treatment Efficacy United States National Institutes of Health Untranslated RNA Work Xenograft procedure career career development chemokine receptor chemotherapy epigenetic regulation experience experimental study fractionated radiation genomic locus improved in vivo knock-down neoplastic cell novel response skills standard care temozolomide therapeutic target therapy resistant transcriptome sequencing treatment response tumor

项目摘要

Project Summary/Abstract Glioblastoma (GBM), the most common primary brain tumor, has a 15 month patient survival due to inevitable recurrence of tumor despite standard treatment – temozolomide chemotherapy (TMZ), radiation, and surgery. It is critical to characterize and target genes modulating GBM chemoresistance. Long noncoding RNAs (lncRNAs) are a novel class of genetic transcripts comprising 80% of the genome. These lncRNAs induce profound alterations in transcriptional regulation and phenotype, including chemotherapy response. Because few GBM lncRNAs have been studied, characterizing new candidates among this abundant and novel class of genes may significantly improve chemotherapeutic efficacy. To assess lncRNAs involved in GBM chemotherapy resistance, we identified linc02454 among the most highly upregulated lncRNAs following TMZ treatment. Knockdown (KD) of linc02454 decreased in vitro GBM cell viability in response to TMZ, while RNA- seq identified CXC-chemokine receptor type 4 (CXCR4) among the most heavily downregulated genes after lncRNA KD. CXCR4 is a well-characterized modulator of GBM chemoresistance, currently targeted in multiple clinical trials. We hypothesize TMZ regulates linc02454 expression and that linc02454 exerts GBM resistance to TMZ through modulation of CXCR4. In this project, we will evaluate the role of linc02454 on GBM response to TMZ treatment. In aim 1, we identify if linc02454 relies upon increased CXCR4 expression, assessing functional interaction via epistasis and CXCR4 rescue, as well as evaluating whether linc02454 directly regulates CXCR4 via Fluorescence in Situ Hybridization. In aim 2, we will assess how TMZ increases linc02454. SA2.1 will perform integrative analysis of lncRNA methylation at DNA loci (including linc02454) and methylation-dependent lncRNA transcription at initial resection/recurrence. SA2.2 will assess chromatin state of linc02454 before and after TMZ. In aim 3, we will assess if linc02454 repression, when combined with TMZ + radiotherapy, improves treatment response, in vitro and in vivo. These aims will examine whether lin02454 meditates GBM response to TMZ via CXCR4 and how linc02454 is regulated by TMZ exposure. We will also test whether linc02454 is a potential therapeutic target in GBM therapy. If successful, this five-year study will provide a more thorough understanding of lncRNA biology in the context of TMZ-induced treatment resistance. With my mentorship group, we have developed a career development plan for tailored training in transcriptional regulation of GBM lncRNAs. The project will be closely supervised through regular one-on-one interaction with primary mentor Dr. Yali Dou, an international expert in chromatin and transcriptional regulation in cancer. NIH-funded co-mentors, Dr. Behnam Badie, a neurosurgeon focusing on GBM biology, and Dr. William Mack also have extensive experience mentoring neurosurgeon-scientists to independence, and have functioned as close mentors. Importantly, by completing this work, I will develop expertise in transcriptional regulation, a critical component in the function of lncRNAs and treatment-induced changes in GBM.

项目摘要/摘要胶质母细胞瘤（GBM）是最常见的原发性脑肿瘤，由于不可避免肿瘤任务标准治疗的复发 - 替莫唑胺化疗（TMZ），辐射和手术。表征和靶向调节GBM化学抗性的基因至关重要。长的非编码RNA （LNCRNA）是一类新的遗传转录本，完成了80％的基因组。这些LNCRNA诱导转录调控和表型的深刻改变，包括化学疗法反应。因为很少有GBM lncrnas研究了，在这一丰富而新颖的阶级中描述了新的候选人基因可能会显着提高化学治疗效率。评估涉及GBM的LNCRNA 化学疗法耐药性，我们在TMZ之后确定了LINC02454的LINC02454 治疗。 LINC02454的敲低（KD）响应TMZ而降低了体外GBM细胞活力，而RNA- SEQ确定了CXC-脱脂受体4型（CXCR4）之后最严重下调的基因 lncrna KD。 CXCR4是GBM化学抗性的特征良好的调节剂，目前针对多个临床试验。我们假设TMZ调节LINC02454表达，而Linc02454具有GBM电阻通过调制CXCR4来进行TMZ。在这个项目中，我们将评估LINC02454在GBM响应中的作用进行TMZ治疗。在AIM 1中，我们确定LINC02454是否依赖于CXCR4表达增加，评估通过上毒和CXCR4救援的功能相互作用，以及评估LINC02454是否直接通过荧光原位杂交调节CXCR4。在AIM 2中，我们将评估TMZ如何增加 Linc02454。 SA2.1将对DNA基因座（包括LINC02454）和甲基化依赖性的lncRNA转录在初始复发/复发时。 SA2.2将评估染色质状态 TMZ之前和之后的Linc02454。在AIM 3中，我们将评估LINC02454表达是否与TMZ结合 +放疗，改善治疗反应，体外和体内。这些目标将检查LIN02454是否通过CXCR4冥想GBM对TMZ的反应，以及LINC02454如何受到TMZ暴露的调节。我们也会测试LINC02454是否是GBM治疗中潜在的治疗靶点。如果成功，这项五年的研究将在TMZ诱导的治疗耐药性的背景下，对LNCRNA生物学提供了更全面的了解。在我的Mentalship Group中，我们制定了一项职业发展计划，用于量身定制的培训 GBM LNCRNA的转录调节。该项目将通过常规一对一的一对一监督与主要导师Yali Dou博士互动在癌症中。 NIH资助的联合官员Behnam Badie博士，专注于GBM生物学的神经外科医生和博士威廉·麦克（William Mack 充当近距离导师。重要的是，通过完成这项工作，我将发展转录方面的专业知识调节，这是LNCRNA功能和治疗诱导的GBM变化的关键成分。