Environmental arsenic, immunoregulation, and viral disease risk

环境砷、免疫调节和病毒性疾病风险

• 批准号: 10452270
• 负责人: JAMES A COFFMAN
• 金额: $ 25.55万
• 依托单位: MOUNT DESERT ISLAND BIOLOGICAL LAB
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-11 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10452270
• 关键词: Acute; Adult; Affect; Animal Model; Anti-Inflammatory Agents; Arsenic; Biological Models; Bronchiectasis; Cells; Cessation of life; Chronic; Clustered Regularly Interspaced Short Palindromic Repeats; Color; Development; Disease; Down-Regulation; Embryo; Endocrine Disruptors; Environmental Exposure; Epigenetic Process; Experimental Models; Exposure to; Failure; Fluorescence Microscopy; Food; Gene Dosage; Gene Expression; Generations; Genes; Genetic; Glucocorticoid Receptor; Glucocorticoids; Goals; Human; Hydrocortisone; Hypothalamic structure; Image; Immune; Immune response; Immune system; Inbreeding; Individual; Infection; Inflammation; Inflammatory; Inflammatory Response; Influenza; Influenza A Virus, H1N1 Subtype; Influenza A virus; Innate Immune Response; Innate Immune System; Knock-out; Label; Larva; Leukocytes; Life; Measures; Mediating; Modeling; Morbidity - disease rate; NF-kappa B; Optics; Pathway interactions; Persons; Pituitary-Adrenal System; Predisposition; Public Health; Puerto Rico; Receptor Signaling; Reporter; Reporting; Repression; Research; Research Project Grants; Resolution; Respiratory Signs and Symptoms; Sampling; Severities; Severity of illness; Signal Pathway; Signal Transduction; Site; Study models; Testing; Therapeutic Intervention; Time; Transgenic Organisms; Viral; Virus; Virus Diseases; Zebrafish; base; cell type; circadian pacemaker; disorder risk; dosage; early life exposure; epidemiological model; flu; glucocorticoid receptor alpha; immunoregulation; in vivo; in vivo Model; influenza infection; intergenerational; knockout gene; lung injury; macrophage; mortality; mortality risk; mutant; neutrophil; novel; pathogen; respiratory virus; response; tool; trafficking; transcription factor; well water

PROJECT SUMMARY / ABSTRACT Arsenic exposure from well water and food is a major public health concern and is associated with increased morbidity and mortality from viral infections. Influenza A virus (IAV) annually infects ~5 million people causing acute respiratory symptoms and up to 646,000 deaths. The severity of viral disease varies between individuals and is likely to involve both genetic and environmental effects on immunoregulation. Epidemiological and animal model studies suggest that early life exposure to arsenic alters the immune response to pathogens, resulting in prolonged or excessive inflammation. Furthermore, available evidence indicates that arsenic is an endocrine disruptor that interferes with the glucocorticoid receptor (GR) signaling pathway, a critical regulator of inflammation, possibly with intergenerational epigenetic effects. However, significant gaps remain in our understanding of how arsenic disrupts GR signaling and promotes inflammation. This exploratory/ developmental research project will use zebrafish as a model system to test the novel hypothesis that arsenic exacerbates viral disease by downregulating the klf9-dependent anti-inflammatory GR signaling pathway. Preliminary studies indicate that Klf9 is a GR-responsive negative regulator of proinflammatory genes, and that basal and cortisol- induced klf9 activity is suppressed in zebrafish embryos exposed to very low levels of arsenic. Zebrafish larvae have a functional innate immune system and are a powerful model to study host-pathogen interactions during systemic or localized influenza (IAV) infection, as trafficking of macrophages and neutrophils to the site of IAV infection can be visualized using live imaging with transgenic lines with fluorescently labeled leukocytes as well as fluorescently labeled viruses. The proposed research will use those tools, as well as GR amd klf9 knockout lines that we recently created using CRISPR, to accomplish two specific aims. The first is to determine if arsenic dysregulates the inflammatory response to IAV infection by suppressing the anti-inflammatory GR-Klf9 signaling pathway, leading to excessive an/or prolonged pro-inflammatory gene expression and failure to resolve the response. This will be accomplished by asking how treatment of zebrafish larvae with arsenic affects expression of klf9 and downstream proinflammatory genes that we have identified as putative targets of Klf9-mediated repression, and assessing the effects of arsenic and klf9 dosage on the response dynamics of inflammatory cells (neutrophils and macrophages) and NF-kB activity following IAV infection. The second specific aim is to determine if arsenic exposure has intergenerational effects on the innate immune response to IAV infection that correlate with aberrant activity of the GR-Klf9 immunoregulatory pathway. To accomplish this, F0 arsenic- or vehicle-exposed wild-type larvae will be raised to adulthood and inbred through 2 generations without further exposure. In larvae from each generation (F1 and F2) we will assess immunoregulatory gene expression and larval survival after IAV infection. The project will elucidate a novel anti-viral immunoregulatory pathway impacted by arsenic, opening an avenue for future research focused on further elucidating the underlying mechanisms.

项目摘要 /摘要 井水和食物的砷暴露是一个主要的公共卫生问题，与增加有关 病毒感染的发病率和死亡率。每年流感病毒（IAV）感染约500万人 急性呼吸道症状和最多646,000人死亡。病毒疾病的严重程度在个体之间有所不同 并且可能涉及遗传和环境对免疫调节的影响。流行病学和动物 模型研究表明，早期生命暴露于砷会改变对病原体的免疫反应，从而改变 长时间或过度炎症。此外，可用证据表明砷是内分泌 干扰糖皮质激素受体（GR）信号通路的破坏者，这是一个关键的调节因子 炎症，可能具有代际表观遗传效应。但是，我们的差距仍然存在 了解砷如何破坏GR信号并促进炎症。这种探索/发展 研究项目将使用斑马鱼作为模型系统，以测试砷加剧病毒的新假设 通过下调KLF9依赖性的抗炎GR信号通路。初步研究 表明KLF9是促炎基因的GR响应性负调节剂，并且基底和皮质醇 - 诱导的KLF9活性在暴露于极低砷水平的斑马鱼胚胎中被抑制。斑马鱼幼虫 具有功能性的先天免疫系统，是研究宿主 - 病原体相互作用的强大模型 全身性或局部流感（IAV）感染，作为巨噬细胞和中性粒细胞的运输到IAV部位 也可以使用带有荧光标记为白细胞的转基因线的实时成像可视化感染 作为荧光标记的病毒。拟议的研究将使用这些工具以及GR AMD KLF9淘汰 我们最近使用CRISPR创建的行，以实现两个具体的目标。首先是确定砷是否 通过抑制抗炎GR-KLF9信号传导，通过抑制对IAV感染的炎症反应失调 途径，导致过度的AN/或延长促炎基因表达和无法解决 回复。这将通过询问用砷对斑马鱼幼虫的治疗如何影响表达来实现 我们已经确定为KLF9介导的推定靶标的KLF9和下游促炎基因 抑制并评估砷和KLF9剂量对炎症细胞反应动力学的影响 （中性粒细胞和巨噬细胞）和IAV感染后的NF-KB活性。第二个具体目的是 确定砷暴露是否对对IAV感染的先天免疫反应有代际影响 与GR-KLF9免疫调节途径的异常活性相关。为此，F0砷或 车辆暴露的野生型幼虫将被抬高到成年，并近交了2代，而无需进一步 接触。在每一代的幼虫中（F1和F2），我们将评估免疫调节基因表达和 IAV感染后的幼虫存活。该项目将阐明一种新型的抗病毒免疫调节途径 由砷，为未来的研究开放途径，重点是进一步阐明基本机制。