The UNC Chapel Hill Superfund Research Program (UNC-SRP)

摘要 本提案作为北卡罗来纳州大学的紧急竞争修订版提交 (UNC)2019冠状病毒病背景下的超级基金研究计划（SRP） 流行病我们专注于无机砷（iAs），这是有毒物质登记署的头号污染物 （ATSDR）污染地球仪的饮用水。iAs对身体的许多器官都是有毒的， 致癌物质、致糖尿病物质、神经毒物和免疫抑制剂。值得注意的是，在北卡罗来纳州的几个州， 饮用水中iAs水平高的县，严重急性呼吸道综合征的患病率 冠状病毒2型（SARS-CoV-2）感染率也很高，表明同时暴露于iAs和SARS-CoV-2可能 发生。这种联合暴露于iAs对SARS-CoV-2感染的易感性和COVID-19严重程度的影响从未 本文研究了本提案的目标是描述iAs暴露与SARS-CoV之间的相互作用， 2感染使用分化的原代人鼻上皮细胞，一个建立的体外模型， 感染，和一种新的人源化hAS 3 MT小鼠品系，其中iAs的代谢和其处置， 代谢物类似于人类。在一个强大的，跨学科的方法，我们将使用不同的 原代人鼻上皮细胞和人源化hAS 3 MT小鼠，以检验慢性鼻粘膜炎的假设。 暴露于iAs会增加对SARS-CoV 2感染的易感性和COVID-19的严重程度。而 结合人类和小鼠模型，我们还将研究性别和遗传背景对 疾病结果。两个新的额外目的包括：（目的1）确定iAs及其代谢产物对 SARS-CoV-2感染分化的原代人鼻上皮细胞;（目的2）表征影响 iAs暴露对hAS 3 MT小鼠SARS-CoV-2 MA感染和COVID-19结局的影响。在目标1中，我们测试 有一种假说认为，接触iAs或其代谢物会增强SARS-CoV-2的易感性和严重性 感染人类鼻上皮细胞。在目标2中，我们将比较免疫应答、病毒滴度、肺 暴露于饮用水中iAs（0、40或400 ppb）并感染 一种适应于老鼠的SARS-CoV-2，最近在美国查佩尔山被制造出来。评估性的作用 和遗传学，我们将使用具有C57 BL/6 NCrl和129 S6/SvEvTac背景的雄性和雌性hAS 3 MT小鼠。 拟议中的研究利用了只有在斯坦福大学才能获得的新颖的实验室模型， 用于研究环境iAs暴露与SARS-CoV 2感染之间的相互作用。拟议 研究解决了理解广泛的环境之间的相互作用的关键需要， 暴露和影响全世界数亿人的大流行感染。

ABSTRACT This proposal is being submitted as an Emergency Competitive Revision for the University of North Carolina (UNC) Superfund Research Program (SRP) in the context of the Coronavirus Disease 2019 (COVID-19) pandemic. We focus on inorganic arsenic (iAs), the #1 contaminant of the Agency for Toxic Substances Registry (ATSDR) contaminating drinking water around the globe. iAs is toxic to many organs in the body, acting as a carcinogen, diabetogen, neurotoxicant, and immunosuppressant. Notably, in several of the North Carolina counties where iAs levels in drinking water are high, the prevalence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is also high, indicating that a co-exposure to iAs and SARS-CoV-2 likely occurs. This co-exposure to iAs on the susceptibility to SARS-CoV-2 infection and COVID-19 severity has never been studied. The goal of this proposal is to characterize the interaction between iAs exposure and SARS-CoV- 2 infection using differentiated primary human nasal epithelial cells, an established in vitro model for respiratory infections, and a novel humanized hAS3MT mouse strain, in which the metabolism of iAs and disposition of its metabolites resemble those in humans. In a strong, transdisciplinary approach, we will use differentiated primary human nasal epithelial cells and a humanized hAS3MT mice to test the hypothesis that chronic exposure to iAs enhances susceptibility to SARS-CoV2 infection and severity of COVID-19. While integrating human and mouse models, we will also examine the role of sex and genetic background on the disease outcome. The two new additional Aims include: (Aim 1) Identify the effects of iAs and its metabolites on SARS-CoV-2 infection in differentiated primary human nasal epithelial cells; and (Aim 2) Characterize the effects of iAs exposure on SARS-CoV-2 MA infection and COVID-19 outcomes in hAS3MT mice. In Aim 1, we test the hypothesis is that exposure to iAs or its metabolites enhances susceptibility to and severity of SARS-CoV-2 infection in the human nasal epithelial cells. In Aim 2, we will compare immune response, viral titer, lung pathology and mortality in hAS3MT mice exposed to iAs in drinking water (0, 40, or 400 ppb) and infected with a mouse-adapted SARS-CoV-2 that has been recently created at UNC Chapel Hill. To assess the role of sex and genetics, we will use male and female hAS3MT mice with C57BL/6NCrl and 129S6/SvEvTac backgrounds. The proposed research utilizes novel laboratory models that are available only at UNC and are uniquely suited for studies of the interaction between environmental iAs exposure and SARS-CoV2 infection. The proposed research addresses a critical need for understanding of the interaction between a widespread environmental exposure and a pandemic infection that affects hundreds of millions of people worldwide.