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

• 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
• 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' 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

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.

项目摘要/摘要 适当的脊椎动物发育需要芳基烃受体（AHR），并被A激活 多种化学物质组包括常见污染物，例如氯化二恶英，多环芳烃 碳氢化合物（PAHS）和共面多氯联苯。在脊椎动物中，暴露于化学物质 在早期阶段激活AHR与不良的出生结果有关 神经发育；但是，AHR下游的信号事件和导致有毒的基因 响应在很大程度上是未知的。我们的长期目标是破译保守的AHR介导的生物学 通过发现和验证AHR涉及的因果基因和信号传导机制的响应 规范的发育和毒理学过程。我们以前鉴定出一种新颖的和系统发育 由多个环境相关的AHR诱导的保守长的非编码RNA（SLINCR） 配体。 SLINCR是AHR诱导的Sox9b的抑制，这是软骨的主调节器 发育，并在AHR介导的出血和软骨缺陷中起因果作用。目标 该建议的是确定AHR调节的lncRNA的发育和毒理学作用， Slincr，在斑马鱼。中心假设是slincr是关键的AHR downstream信号传导因子 增强对各种异种化学化学物质的不良发育反应。我们的理由是 了解SLINCR的功能以及与之相互作用的其他因素如何加深理解 AHR信号，可能会发现与AHR相关疾病的人类治疗靶标。我们 将通过执行损失和功能性研究来确定体内作用来检验我们的假设 AHR规范的发育和毒理学过程中的SLINCR（AIM 1）。了解slincr 结构功能与AHR信号的关系，我们将执行结构映射实验，并 识别SLINCR相互作用的蛋白质和全基因组靶标（AIM 2）。我们将使用计算 将目标1和2的发现集成到AHR基因调节网络的构建中的方法， 在AHR调节的生物过程中建立对SLINCR的系统级别的理解（AIM 3）。这 贡献很重要，因为它将揭示AHR激活对SLINCR表达的功能影响 并提供一个新的范式来理解依赖AHR的细胞反应。拟议的研究 当我们采用新的环境健康科学领域的尖端技术时，具有创新性。 我们的建议还将有助于描述类似二恶英的AHR配体的新型不良结果途径（AOP） 结合了多个生物学水平，包括分子结构，分子相互作用，共同 调控基因，信号网络，组织发育和全动物表型。这些研究会 充当LNCRNA增强生物学的系统功能表征的模板 疾病。