Defining the developmental and toxicological roles of the AHR-regulated Sox9 lncRNA in zebrafish

定义 AHR 调节的 Sox9 lncRNA 在斑马鱼中的发育和毒理学作用

基本信息

批准号：
10088447
负责人：
Robyn L Tanguay
金额：
$ 43.27万
依托单位：
OREGON STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-02-01 至 2022-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10088447
关键词：
5&apos Untranslated Regions AHR gene Affinity Chromatography Agonist Animals Aromatic Hydrocarbons Aromatic Polycyclic Hydrocarbons Aryl Hydrocarbon Receptor Autoimmune Binding Biological Biological Process Biology Blood Vessels Cartilage Chemical Exposure Chemicals DNA Data Defect Development Dioxins Disease Environmental Health Event Exposure to Gene Expression Genes Genetic Transcription Genome Goals Grant Health Hemorrhage Human Impairment Knowledge Lead Life Ligands Mass Spectrum Analysis Mediating Modeling Molecular Molecular Structure Mus Orthologous Gene Outcome Pathway interactions Phenotype Phylogenetic Analysis Play Polychlorinated Biphenyls Process Proteins Proteome RNA Receptor Activation Receptor Signaling Regulator Genes Repression Research Role Signal Transduction Societies Structure Structure-Activity Relationship System Testing Tetrachlorodibenzodioxin Tissues Untranslated RNA Validation Vertebrates Xenobiotics Zebrafish adverse birth outcomes adverse outcome angiogenesis aryl hydrocarbon receptor ligand base cardiogenesis cartilage development causal variant chemical group comparative developmental toxicology environmental chemical experimental study exposed human population gain of function gene repression genome-wide human disease in silico in vivo innovation neurodevelopment new technology next generation sequencing novel pollutant receptor reproductive development response therapeutic target

项目摘要

PROJECT SUMMARY/ABSTRACT The aryl hydrocarbon receptor (AHR) is required for proper vertebrate development and is activated by a diverse group of chemicals including common pollutants such as chlorinated dioxins, polycyclic aromatic hydrocarbons (PAHs), and coplanar polychlorinated biphenyls. In vertebrates, exposure to chemicals that activate the AHR during early life stages is associated with adverse birth outcomes and impaired neurodevelopment; however, the signaling events downstream of AHR and the genes contributing to toxic responses remain largely unknown. Our long-term goal is to decipher conserved AHR-mediated biological responses via discovery and validation of the causal genes and signaling mechanisms involved in AHR regulated developmental and toxicological processes. We previously identified a novel and phylogenetically conserved long non-coding RNA (slincR) that is induced by multiple environmentally relevant AHR ligands. SlincR is required for the AHR-induced repression of sox9b, a master regulator of cartilage development, and plays a causal role in AHR-mediated hemorrhaging and cartilage defects. The objective of this proposal is to identify the developmental and toxicological roles of the AHR-regulated lncRNA, slincR, in zebrafish. The central hypothesis is that slincR is a critical AHR-downstream signaling factor potentiating adverse developmental responses to diverse xenobiotic chemicals. Our rationale is that understanding how slincR functions and what other factors it interacts with will deepen the understanding of AHR-signaling with the potential to uncover human therapeutic targets for AHR-related disease. We will test our hypothesis by performing loss- and gain-of-function studies to determine the in vivo role of slincR in AHR regulated developmental and toxicological processes (Aim 1). To understand the slincR structure-function relationship with AHR signaling, we will perform structure-mapping experiments and identify the slincR interacting proteins and genome-wide targets (Aim 2). We will use computational approaches to integrate the findings of Aims 1 & 2 into the construction of an AHR gene regulatory network, establishing a systems-level understanding of slincR in AHR-regulated biological processes (Aim 3). This contribution is significant as it will unravel the functional impact of AHR activation on slincR expression and provide a new paradigm for understanding AHR-dependent cellular responses. The proposed research is innovative as we employ cutting-edge technologies new to the field of environmental health sciences. Our proposal will also help to describe a novel adverse outcome pathway (AOP) for dioxin-like AHR ligands that incorporate multiple biological levels including molecular structure, molecular interactions, co- regulated genes, signaling networks, tissue development and whole animal phenotypes. These studies will serve as a template for the systematic functional characterization of lncRNA potentiating biology and disease.

项目概要/摘要芳基碳氢化合物受体 (AHR) 是脊椎动物正常发育所必需的，并由多种化学物质，包括常见污染物，如氯化二恶英、多环芳香族化合物碳氢化合物 (PAH) 和共面多氯联苯。对于脊椎动物来说，接触化学物质会在生命早期阶段激活 AHR 与不良出生结果和受损有关神经发育；然而，AHR 下游的信号事件和导致毒性的基因对此的反应很大程度上仍不得而知。我们的长期目标是破译保守的 AHR 介导的生物通过发现和验证 AHR 中涉及的因果基因和信号机制来做出反应调节发育和毒理学过程。我们之前发现了一种新颖且系统发育的由多个环境相关的 AHR 诱导的保守长非编码 RNA (slincR) 配体。 SlincR 是 AHR 诱导的软骨主要调节因子 sox9b 抑制所必需的发育，并在 AHR 介导的出血和软骨缺陷中起因果作用。目标该提案的目的是确定 AHR 调节的 lncRNA 的发育和毒理学作用， slincR，在斑马鱼中。中心假设是 slincR 是一个关键的 AHR 下游信号传导因子加剧对多种外源化学物质的不良发育反应。我们的理由是了解 slincR 的功能以及它与哪些其他因素相互作用将加深理解 AHR 信号传导的研究有可能发现 AHR 相关疾病的人类治疗靶点。我们将通过进行功能丧失和获得功能研究来检验我们的假设，以确定其体内作用 AHR 中的 slincR 调节发育和毒理学过程（目标 1）。了解 slincR 与AHR信号传导的结构-功能关系，我们将进行结构映射实验并识别 slincR 相互作用蛋白和全基因组靶标（目标 2）。我们将使用计算将目标 1 和 2 的发现整合到 AHR 基因调控网络的构建中的方法，建立对 AHR 调节的生物过程中 slincR 的系统级理解（目标 3）。这贡献意义重大，因为它将揭示 AHR 激活对 slincR 表达的功能影响并为理解 AHR 依赖性细胞反应提供新的范例。拟议的研究我们采用环境健康科学领域的尖端技术，因此具有创新性。我们的提案还将有助于描述二恶英类 AHR 配体的新型不良后果途径 (AOP) 结合了多个生物学水平，包括分子结构、分子相互作用、共调控基因、信号网络、组织发育和整个动物表型。这些研究将作为 lncRNA 增强生物学系统功能表征的模板疾病。