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: 10589936
• 负责人: JAMES A COFFMAN
• 金额: $ 19.48万
• 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 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10589936
• 关键词: Acute; Adrenal Glands; 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; Gene Expression Profiling; Generations; Genes; Genetic; Glucocorticoid Receptor; Glucocorticoids; Goals; Heterozygote; Human; Hydrocortisone; Hypothalamic structure; Image; Immune; Immune response; Immune system; Inbreeding; Individual; Infection; Inflammation; Inflammatory; Inflammatory Response; Influenza A Virus, H1N1 Subtype; Influenza A virus; Innate Immune Response; Innate Immune System; Knock-out; Label; Larva; Leukocytes; Life; Macrophage; Measures; Mediating; Modeling; Morbidity - disease rate; NF-kappa B; Optics; Pathway interactions; Persons; Pituitary Gland; 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; Visualization; Zebrafish; cell type; circadian pacemaker; disorder risk; dosage; dynamic system; early life exposure; epidemiological model; flu; immunoregulation; in vivo; in vivo Model; influenza infection; intergenerational; knockout gene; lung injury; mortality; mortality risk; mutant; neutrophil; novel; pathogen; respiratory virus; response; tool; trafficking; transcription factor; transmission process; 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.

项目概要/摘要 井水和食物中的砷暴露是一个主要的公共卫生问题，并且与砷的增加有关。 病毒感染的发病率和死亡率。甲型流感病毒 (IAV) 每年感染约 500 万人，导致 急性呼吸道症状和多达 646,000 人死亡。病毒性疾病的严重程度因人而异 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，生命早期接触砷会改变对病原体的免疫反应，导致 长期或过度的炎症。此外，现有证据表明砷是一种内分泌激素。 干扰糖皮质激素受体 (GR) 信号通路的干扰物，糖皮质激素受体 (GR) 信号通路是糖皮质激素受体 (GR) 的关键调节因子 炎症，可能具有代际表观遗传效应。然而，我们在这方面仍存在重大差距 了解砷如何破坏 GR 信号传导并促进炎症。这种探索性/发展性 研究项目将使用斑马鱼作为模型系统来测试砷加剧病毒的新假设 通过下调 klf9 依赖性抗炎 GR 信号通路来治疗疾病。初步研究 表明 Klf9 是促炎基因的 GR 响应性负调节因子，并且基础和皮质醇- 在暴露于极低水平砷的斑马鱼胚胎中，诱导的 klf9 活性受到抑制。斑马鱼幼虫 具有功能性的先天免疫系统，是研究宿主与病原体相互作用的强大模型 全身或局部流感 (IAV) 感染，巨噬细胞和中性粒细胞运输至 IAV 部位 也可以使用带有荧光标记白细胞的转基因系的实时成像来可视化感染 如荧光标记的病毒。拟议的研究将使用这些工具以及 GR amd klf9 淘汰赛 我们最近使用 CRISPR 创建了一些细胞系，以实现两个特定目标。首先是确定是否砷 通过抑制抗炎 GR-Klf9 信号传导失调对 IAV 感染的炎症反应 途径，导致过度和/或延长促炎基因表达并未能解决问题 回复。这将通过询问砷处理斑马鱼幼虫如何影响表达来完成 klf9 和下游促炎基因，我们已将其确定为 Klf9 介导的推定靶标 抑制，并评估砷和 klf9 剂量对炎症细胞反应动力学的影响 IAV 感染后（中性粒细胞和巨噬细胞）和 NF-kB 活性。第二个具体目标是 确定砷暴露是否对 IAV 感染的先天免疫反应具有代际影响 与 GR-Klf9 免疫调节途径的异常活性相关。为了实现这一点，F0 砷-或 暴露于车辆的野生型幼虫将被饲养至成年并近交2代，无需进一步 接触。在每一代（F1 和 F2）的幼虫中，我们将评估免疫调节基因表达和 IAV 感染后幼虫的存活率。该项目将阐明受影响的新型抗病毒免疫调节途径 通过砷，为未来的研究开辟了一条途径，重点是进一步阐明潜在的机制。