Interactions between metabolism, transport, and toxicity of benzalkonium chlorides

项目摘要 苯扎氯铵（BAC）是广泛用于消毒产品、医疗产品 消费品和食品加工业，这表明人类可能长期和系统地接触， 通过各种途径向BAC提供技术支持。我们的初步研究发现，100个随机胡- 人血浆样品含有可检测水平的BAC，表明BAC确实被吸收。正在进行的 COVID-19大流行导致含BAC消毒剂的使用大幅增加，这令人担忧 鉴于越来越多的证据表明， BAC和BAC诱导的啮齿动物胆固醇和脂质稳态破坏。然而，缺乏 了解BAC在人体内的代谢、转运和生物学后果。我们的目标是 表征BAC的代谢和转运途径及其对BAC肾毒性的影响。 先前在大鼠中进行的研究表明，BAC蓄积至 口服后肾脏中的最高水平，我们的初步研究表明，BAC发挥强大的细胞毒性， 在3D“肾芯片”微生理系统（MPS）中的功能。最近，我们报道了BAC是代谢- 在体外被人细胞色素P450（CYP）亚型CYP 2D 6和CYP 4s分解。此外，我们发现， BAC由人有机阳离子转运蛋白（hOCT）主动转运。因为CYP 2D 6、CYP 4s和 hOCT是高度多态性，具有非常不同的蛋白质活性，我们假设BAC在 肾脏的功能依赖于肝脏和肾脏中BAC代谢和转运蛋白的活性。 在目标1中，我们将表征体外BAC的代谢和转运途径，包括二级代谢和转运途径。 通过β-氧化和葡萄糖醛酸化的禁忌症和通过hOCT、人多药和毒素外排前体的转运 蛋白和P-糖蛋白。目的2：评价BAC对人近曲小管的肾毒性 3D集成肝肾“器官芯片”MPS中的上皮细胞。一个综合的立体组学，脂质组学， 转录组学方法将用于系统地评估BAC的毒性和生物活性。在 目的3，我们将评估BAC暴露水平及其与人类脂质和肾损伤生物标志物的相关性。 人类，以及遗传变异对BAC代谢和处置的影响。其意义 该项目在于，它将首次解决代谢，运输和毒性方面的知识差距， 人体内的BAC阐明CYP和/或转运蛋白活性降低对BAC毒性的贡献- 这将使我们能够识别这些蛋白质中具有遗传变异的高危人群。的 所获得的知识还可以告知联邦机构如何设定更适当的接触限制。的 该项目的创新之处在于a）通过异生物质- B）使用整合的肝肾MPS评估以下物质的毒理学后果： 异生物质，以及c）用于严格系统毒理学研究的综合组学。

Project Summary Benzalkonium chlorides (BACs) are widely used antimicrobials in disinfecting products, medical products, consumer products, and food processing industries, suggesting humans may be exposed chronically and sys- temically to BACs through a variety of routes. Our preliminary study found that close to 50 of 100 random hu- man plasma samples contain detectable levels of BACs, suggesting BACs are indeed absorbed. The ongoing COVID-19 pandemic has led to greatly increased use of BAC-containing disinfectants, which is concerning given accumulating evidence in respiratory, developmental, reproductive, and neurological toxicities inflicted by BACs and BAC-induced disruption of cholesterol and lipid homeostasis in rodents. However, there is a lack of knowledge on the metabolism, transport, and biological consequences of BACs in humans. Our goal is to characterize the pathways of metabolism and transport of BACs and their impact on nephrotoxicity of BACs. The potential for nephrotoxicity is supported by previous studies in rats showing that BACs accumulate to the highest level in the kidney after oral intake and our preliminary studies showing that BACs exert potent cytotox- icity in a 3D “kidney-on-a-chip” microphysiological system (MPS). Recently, we reported that BACs are metab- olized by human cytochrome P450 (CYP) isoforms CYP2D6 and CYP4s in vitro. Furthermore, we found that BACs are actively transported by human organic cation transporters (hOCTs). Because CYP2D6, CYP4s, and hOCTs are highly polymorphic with greatly varying protein activities, we hypothesize that toxicities of BACs in kidney are dependent on the activities of BAC-metabolizing and transporting proteins in both liver and kidney. In Aim 1, we will characterize pathways of metabolism and transport of BACs in vitro, including secondary me- tabolism by β-oxidation and glucuronidation and transport by hOCTs, human multidrug and toxin extrusion pro- teins, and P-glycoprotein. In Aim 2, we will evaluate nephrotoxicity induced by BACs in human proximal tubule epithelial cells in 3D integrated liver-kidney “organs-on-chips” MPS. An integrated sterolomics, lipidomics, and transcriptomics approach will be used to systemically assess the toxicity and biological activities of BACs. In Aim 3, we will assess BAC exposure levels and their correlation with lipid and kidney injury biomarkers in hu- mans, as well as the impact of genetic variations on BAC metabolism and disposition. The significance of this project lies in that it will, for the first time, address the knowledge gap in metabolism, transport, and toxicity of BACs in humans. Elucidation of the contribution of reduced activities in CYPs and/or transporters to BAC tox- icity would enable us to identify high-risk human population with genetic variations in these proteins. The gained knowledge could also inform federal agencies on setting more appropriate exposure limitations. The innovation of this project lies in a) a novel example of gene-environment interaction through xenobiotic- processing proteins, b) the use of an integrated liver-kidney MPS to assess the toxicological consequences of xenobiotics, and c) integrated omics for rigorous systems toxicology studies.