Conserved AHR-dependent control of Sox9 long non coding RNA expression

Sox9 长非编码 RNA 表达的保守 AHR 依赖性控制

基本信息

批准号：
8884471
负责人：
Robyn L Tanguay
金额：
$ 21.81万
依托单位：
OREGON STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-05-01 至 2017-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8884471
关键词：
Address Affect Animals Aryl Hydrocarbon Receptor Benz(a)Anthracenes Binding Bioinformatics Biological Process Chemicals Clustered Regularly Interspaced Short Palindromic Repeats Complex Congenital Abnormality Coupled Data Development Disease Down-Regulation Embryonic Development Environmental Pollution Enzymes Event Exploratory/Developmental Grant Exposure to Fishes Gene Expression Gene Targeting Genes Genetic Transcription Genomics Goals High-Throughput Nucleotide Sequencing Homeostasis Human Human Development Immune system In Situ Hybridization Inflammation Knock-out Knowledge Ligands Mammals Maps Mediating Messenger RNA Modeling Molecular Toxicology Mus Organ Outcome Pathway interactions Positioning Attribute Quinones RNA RNA Binding RNA analysis Receptor Activation Receptor Signaling Regulation Regulatory Element Relative (related person)Reporter Reporting Research Role Signal Pathway Signal Transduction Site Structure System Testing Tetrachlorodibenzodioxin Therapeutic Uses Toxic Environmental Substances Toxic effect Toxicology Transcript Transcriptional Regulation Treatment Efficacy Untranslated RNA Xenobiotics Zebrafish activating transcription factor adverse outcome aryl hydrocarbon receptor ligand benzanthracene beta catenin cell type craniofacial developmental toxicity gene repression genome editing genome-wide analysis human disease immune function improved in vivo innovation loss of function men novel novel strategies overexpression progenitor promoter public health relevance reproductive research study response tool transcription factor transcriptome sequencing tumor

项目摘要

DESCRIPTION (provided by applicant): Since the discovery of the Aryl Hydrocarbon Receptor (AHR) 20 years ago, it is clear that this ligand activated transcription factor is intimately integrated with numerous signaling pathways that modulate biological processes. Exposure to AHR ligands profoundly impacts embryonic development, immune function, and promote tumor development. It has been implicated in multiple human diseases. Furthermore, selective AHR modulators are now being developed for therapeutic uses to suppress inflammation. Elucidating the complex events immediately downstream of AHR activation is critical to parse toxicity and improve therapeutic efficacy. The bulk of research into the downstream AHR targets has focused on mRNA induction, despite ample evidence that numerous transcripts are repressed by AHR activation. One key down-regulated target is sox9, a transcription factor that controls specification and differentiation of numerous progenitor and differentiated cell types. Sox9 haploinsufficiency causes a multitude of severe developmental problems in humans, but the mechanisms governing sox9 transcription remain unsolved. We and others have reported that TCDD leads to a rapid down regulation of sox9 transcription in an AHR dependent mechanism in developing mammals and fish, which is manifest through craniofacial and reproductive effects. Understanding the mechanisms of AHR- dependent transcriptional repression of sox9b has stymied us for six years, until we completed an unbiased deep RNA-seq study that identified a novel long non-coding RNA (lncRNA) transcript induced by developmental exposure to AHR ligands. This lncRNA is directly adjacent to the sox9b gene. Importantly the lncRNA-sox9b genomic relationship is conserved in mice and humans. We hypothesize a new adverse outcome pathway whereby AHR activation induces sox9b-lncRNA which in turn down-regulates sox9b transcription. Using AHR2-null fish, we confirmed that the sox9b-lncRNA induction is absolutely AHR2-dependent. We also identified AHR regulatory elements in the promoters of both zebrafish and mammalian lncRNA-sox9b genes. To test the hypothesis that the conserved sox9-lncRNA is a direct AHR target gene that upon AHR activation represses sox9b leading to target organ-specific toxicity, we will complete two Specific Aims: 1) We will determine the temporal and spatial expression of zebrafish sox9b-lncRNA relative to the sox9b mRNA, determine its binding to the sox9b promoter in vivo, and identify other potential targets of the sox9b-lncRNA by genome-wide analysis; 2) Using sox9b-lncRNA gain and loss-of-function studies, we will also define the role of sox9b-lncRNA in AHR-dependent developmental toxicity. Collectively, these studies are ideal for the R21 mechanism to build the new tools need to probe the role of lncRNAs in toxicity and develop a new approach for unraveling an unexpected layer of complexity affecting adverse outcome pathways.

描述（由适用提供）：由于20年前发现了芳基烃受体（AHR），因此很明显，该配体激活的转录因子与调节生物学过程的许多信号通路紧密整合。暴露于AHR配体会深刻影响胚胎发育，免疫功能并促进肿瘤发育。它与多种人类疾病有关。此外，现在正在开发选择性的AHR调节剂来抑制注射。立即阐明AHR激活下游的复杂事件对于解析毒性和提高治疗效果至关重要。对下游AHR靶标的大量研究集中在mRNA诱导上，目的地充分的证据表明，AHR激活反映了许多转录本。一个关键的下调目标是SOX9，它是控制众多祖细胞和分化细胞类型的规范和分化的转录因子。 Sox9单倍不足造成了人类的多种严重发育问题，但是控制SOX9转录的机制仍未解决。我们和其他人报告说，TCDD导致在开发哺乳动物和鱼类中AHR依赖机制中Sox9转录的快速调节，这通过颅面和生殖作用显现出来。了解Sox9b的AHR依赖性转录表示的机制使我们陷入了六年的困扰，直到我们完成了一项无偏见的深RNA-Seq研究，该研究鉴定出通过发育暴露于AHR配体引起的新型长期非编码RNA（LNCRNA）。重要的是，lncRNA-SOX9B基因组关系在小鼠和人类中是保守的。我们假设一种新的不良结果途径，通过该途径AHR激活诱导SOX9B-LNCRNA，反过来降低了SOX9B转录。使用AHR2-NULL鱼，我们证实了Sox9b-lncRNA诱导绝对依赖于AHR2。我们还确定了斑马鱼和哺乳动物lncRNA-SOX9B基因的启动子中的AHR调节元件。 To test the hypothesis that the configured sox9-lncRNA is a direct AHR target gene that upon AHR activation reflects sox9b leading to target organ-specific toxicity, we will complete two Specific Aims: 1) We will determine the temporary and spatial expression of zebrafish sox9b-lncRNA relative to the sox9b mRNA, determine its binding to the sox9b promoter in vivo, and identify other potential targets of the通过全基因组分析，SOX9B-LNCRNA； 2）使用SOX9B-LNCRNA的增益和功能丧失研究，我们还将定义SOX9B-LNCRNA在AHR依赖性发育毒性中的作用。总的来说，这些研究是R21机制构建新工具的理想选择，以探测LNCRNA在毒性中的作用，并开发一种新方法，以揭示影响不良结果途径的意外复杂性层。