KDM5B Mediates Cell Survival in MYC-Dependent T-ALL

KDM5B 介导 MYC 依赖性 T-ALL 中的细胞存活

基本信息

批准号：
10238119
负责人：
Daniel F Liefwalker
金额：
$ 18.29万
依托单位：
OREGON HEALTH & SCIENCE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-17 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10238119
关键词：
Acute T Cell Leukemia Apoptotic BHLH Protein Binding Binding Sites Biological Bromodomain CRISPR/Cas technology Cancer Center Cell Death Cell Line Cell Survival Cells Cellular biology Chromatin Clinical Clinical Treatment Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Complex Computer Models Data Data Set Development Emu species Epigenetic Process Event Family Gene Expression Profiling Genes Genetic Transcription Genome Hematologic Neoplasms Human Human Cell Line Image KDM5B gene Knock-out Lead Lymphoma Lymphoma cell Malignant Neoplasms Mediating Mediator of activation protein Mentors Modeling Mus Mutagenesis Mutation Oncogenes Oncogenic Pathway interactions Promoter Regions Proto-Oncogenes Regulation Relapse Repression Research Role Sampling Signal Transduction Systems Biology Testing Tetanus Helper Peptide Therapeutic Time Transcription Repressor Transgenic Model Transgenic Organisms Tumor Suppressor Proteins Up-Regulation Xenograft procedure acute T-cell lymphoblastic leukemia cell c-myc Genes cancer cell cancer initiation cancer subtypes chromatin immunoprecipitation comparative experimental study gene repression genetically modified cells genome-wide histone demethylase in vivo inhibitor/antagonist member mouse model novel novel therapeutic intervention novel therapeutics oncogene addiction overexpression programs response therapeutic target transcription factor transcriptome sequencing treatment strategy tumor tumor progression tumorigenesis tumorigenic vector

项目摘要

Project Summary/Abstract Cancer is a complex landscape of aberrant cell signaling programs, initiated by oncogenes. The causal oncogene drives the preponderance of accumulated mutations that enable tumorigenesis. Cancers arising this way can be reliant on the initiating oncogene, a term known as oncogene addiction. The proto-oncogene c- MYC is a transcription factor that regulates much of the genome and is deregulated in many cancers. The transgenic Eµ-tTA/Tet-O-MYC mouse model of T-cell acute lymphoblastic leukemia (T-ALL), allows for modulation of c-MYC expression (ON vs OFF). When c-MYC is expressed T-ALL progression is observed, and when c-MYC expression is revoked tumor regression occurs. Using cells and microarray data from this model we employed a nested effects model (NEM) that infers hierarchical relationships anchored to master transcription factors that govern critical aspects of cell biology. We identified a critical node governed by a class of histone demethylases influencing transcriptional availability across the genome. KDM5B/JARID1b is known as a transcriptional repressor, and is downregulated in the T- ALL model when c-MYC is overexpressed. Using CRISPR/Cas9 mediated mutagenesis to disrupt KDM5B expression, we observed a potent reduction in cell death when c-MYC expression is abrogated; suggesting KDM5B mediates cell death responses in T-ALL. The central hypothesis is that KDM5B acts as a tumor suppressor in c-MYC-dependent T-ALL. We propose the following three aims to test this hypothesis. Aim 1 will determine if c-MYC directly suppresses KDM5B expression and whether this is critical in c- MYC-dependent T-ALL. Chromatin Immunoprecipitation (ChIP) of c-MYC at the promoter regions of KDM5B will identify whether direct regulation is required. Aim 2 will identify the mechanism through which KDM5B regulates cell survival in c-MYC-dependent T-ALL. Using RNA-seq and ChIP-seq approaches we will uncover critical c-MYC and KDM5B regulated gene programs and dissect the pathways unique to KDM5B. Aim 3 will discover how KDM5B influences tumor development in vivo. Genetically modified cell lines that over-express KDM5B as well as KDM5B knockout cells will be injected into syngeneic hosts to determine KDM5B influence on tumor development in vivo. Preliminary data suggests that lymphoma cells treated with the bromodomain inhibitor JQ1 undergo cell death only if c-MYC is downregulated and KDM5B is upregulated. Human T-ALL cell lines responsive to JQ1 will be injected into NOD-SCIDIL-2Rg-/- (NSG) mice and tumor progression will be tracked by bioluminescent imaging (BLI). Additionally, primary human lymphoma samples will be injected into NDG mice and treated with the BET inhibitor JQ1. These studies are the basis of an independent research program and demonstrate a novel paradigm in understanding how c-MYC promotes tumor development through repression of a tumor suppressive epigenetic landscape regulated by KDM5B, and identify therapeutic options for treating c-MYC-induced T-ALL. !

项目摘要/摘要癌症是由癌基因引发的异常细胞信号程序的复杂景观。因果关系致癌基因驱动了能够造成肿瘤发生的累积突变的优势。癌症引起了这种情况方式可以依赖于癌基因（一种称为癌基因成瘾的术语）。原始癌基因C- MYC是调节大部分基因组的转录因子，并在许多癌症中受到管制。 T细胞急性淋巴细胞白血病（T-ALL）的转基因Eµ-TTA/TET-O-MYC小鼠模型允许 C-MYC表达的调制（ON VS OFF）。当C-Myc表达T-ALL进展时，当C-Myc表达被吊销时，会发生肿瘤回归。使用该模型的单元格和微阵列数据，我们采用了嵌套效应模型（NEM）分层关系锚定为控制细胞生物学关键方面的主要转录因子。我们确定了由一类组蛋白脱甲基酶影响转录的关键节点跨基因组。 kdm5b/jarid1b被称为转录复制品，在t-中被下调当C-Myc过表达时，所有模型。使用CRISPR/CAS9介导的诱变破坏KDM5B 表达，我们观察到当C-Myc表达废除时细胞死亡的潜在减少。建议 KDM5B介导T-ALL中的细胞死亡反应。中心假设是KDM5B充当肿瘤 C-Myc依赖性T-ALL中的抑制剂。我们提出以下三个旨在检验这一假设的目的。 AIM 1将确定C-MYC是否直接抑制KDM5B的表达，以及在C-中是否至关重要 MYC依赖T-All。 C-Myc的染色质免疫沉淀（芯片）在KDM5B的启动子区域将确定是否需要直接法规。 AIM 2将确定KDM5B的机制调节C-MYC依赖性T-ALL中的细胞存活。使用RNA-seq和Chip-seq方法，我们将发现关键的C-MYC和KDM5B调节基因程序，并剖析KDM5B独有的途径。目标3意志发现KDM5B如何影响体内肿瘤的发展。过表达的转基因细胞系 KDM5B以及KDM5B基因敲除细胞将被注入合成宿主，以确定KDM5B的影响关于体内肿瘤的发育。初步数据表明用溴结构域处理的淋巴瘤细胞抑制剂JQ1仅在下调C-MYC并更新KDM5B时才会发生细胞死亡。人类T-All 对JQ1响应的细胞系将注入nod-scidil-2rg - / - （NSG）小鼠，肿瘤进展将为通过生物发光成像（BLI）跟踪。另外，将原发性人淋巴瘤样品注入 NDG小鼠并用BET抑制剂JQ1治疗。这些研究是独立研究计划的基础，并证明了新颖的范式了解C-MYC如何通过反射肿瘤表观遗传来促进肿瘤的发展由KDM5B调节的景观，并确定治疗C-Myc诱导的T-all的治疗选择。呢