Mechanisms underlying metal nanoparticle-induced lung injury and fibrosis

金属纳米颗粒诱导肺损伤和纤维化的机制

• 批准号: 10162656
• 负责人: Qunwei Zhang
• 金额: $ 39万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10162656
• 关键词: Adaptor Signaling Protein; Address; Alveolar Macrophages; Antibodies; Apoptosis; Asbestos; Asthma; Attenuated; Biological; Bronchoalveolar Lavage Fluid; CASP1 gene; CRISPR/Cas technology; Caspase; Cells; Collagen; Cosmetics; Development; Electronics; Epithelial Cells; Exposure to; Family; Fibroblasts; Fibrosis; Gelatinase A; Gelatinase B; Generations; Health; In Vitro; Industrialization; Industry; Inflammasome; Inflammatory; Injury; Interleukin-1; Interleukin-1 Receptors; Interleukin-1 beta; Knockout Mice; Lead; Lesion; Lung; Lung Inflammation; Lung diseases; Malignant neoplasm of lung; Matrix Metalloproteinases; Measures; Medical; Medicine; Metal exposure; Metal fever; Metals; Mitochondria; Modeling; Multiprotein Complexes; Mus; NADPH Oxidase; Normal Range; Occupational; Pharmacology; Play; Potassium; Proteins; Pulmonary Fibrosis; Regulation; Role; Silicon Dioxide; System; Testing; Tissue Inhibitor of Metalloproteinases; Toxic effect; Transition Elements; Wild Type Mouse; anakinra; base; exposed human population; in vivo; inhibitor/antagonist; interest; interleukin-1beta-converting enzyme inhibitor; knock-down; lung injury; macrophage; member; monocyte; nano; nanomaterials; nanoparticle; particle; recruit; response; titanium dioxide; tool

Metal nanoparticles have been widely used in cosmetics, medicine, electronics, and industry, and occupational or non-occupational exposure to metal nanoparticles is growing. In this proposal, we have selected several transition metal nanoparticles (Nano-Co, Nano-Ni, and Nano-TiO2) as `model' metal nanoparticles to examine their ability to induce pulmonary injury and fibrosis and the potential underlying mechanisms involved. An inflammasome is a multiprotein complex that serves as a platform for caspase-1-dependent proteolytic maturation and secretion of interleukin-1β (IL-1β). The central component of an inflammasome is a member of the NLRP family, and this protein associates with the adaptor protein ASC, which in turn recruits pro-inflammatory caspase precursors (such as procaspase-1). Among a number of inflammasomes, the NLRP3 inflammasome is the most extensively studied. Our working hypothesis is that exposure to metal nanoparticles will cause activation and/or dysregulation of the inflammasome and IL-1β secretion in alveolar macrophages (AMs), lung epithelial cells, and lung fibroblasts, which will cause dysregulation of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs), initiating and promoting metal nanoparticle- induced pulmonary injury and fibrosis. This project will use both in vitro and in vivo systems to address the following specific aims: (1) Determine the role of inflammasome activation in IL-1β secretion induced by metal nanoparticles in vitro and in vivo. We will identify whether activation of the inflammasome is involved in metal nanoparticle-induced IL-1β secretion in alveolar macrophages (AMs), lung epithelial cells, and lung fibroblasts by: (1) using ac-YVAD-cmk, a particular inhibitor of caspase-1; (2) knocking-down one of the inflammasome components such as NLRP3, ASC, or caspase-1 by using CRISPR/Cas9 technology; and (3) using NLRP3 or ASC knock-out mice. We will then determine whether NADPH oxidase- and/or mitochondria- dependent ROS generation and potassium efflux are involved in metal nanoparticle-induced inflammasome activation and IL-1β secretion. We will also measure IL-1β secretion in mice exposed to metal nanoparticles. (2) Examine the role of IL-1β in the alteration of MMPs and TIMPs expression and activity in lung cells exposed to metal nanoparticles. While IL-1β is an inducer for MMP-2 and MMP-9 activity, it is unclear how it regulates MMPs and TIMPs with exposure to metal nanoparticles. To test the role of the inflammasome and IL- 1β in the regulation of MMPs and TIMPs, the strategies in Aim 1 will be used to inhibit inflammasome function, and strategies to inhibit IL-1 function will be applied by using: (1) the pharmacologic IL-1β inhibitor; (2) anti-IL- 1β antibody; and (3) IL-1RI-/- mice that will not respond to IL-1β. After exposure to metal nanoparticles, MMP-2, MMP-9 and TIMPs expression and activity will be determined. (3) Investigate the role of inflammasome activation in metal nanoparticle-induced lung injury and fibrosis in vivo. We will first investigate whether exposure to metal nanoparticles will cause lung fibroblasts to produce more collagen. Then the role of the inflammasome in metal nanoparticle-induced lung fibrosis will be explored by short- and long-term exposure of mice to metal nanoparticles. We will use the strategies in Aim 1 and 2 to inhibit inflammasome and IL-1β function to investigate the role of the inflammasome and IL-1β in metal nanoparticle-induced lung fibrosis.

金属纳米颗粒已被广泛用于化妆品，医学，电子和工业以及职业 或非占用对金属纳米颗粒的暴露正在增长。在此提案中，我们选择了几个 过渡金属纳米颗粒（纳米-CO，纳米-NI和纳米-TIO2）作为“模型”金属纳米颗粒进行检查 它们诱导肺部损伤和纤维化的能力以及潜在的潜在机制。一个 炎症体是一种多蛋白质复合物，可作为caspase-1依赖性蛋白水解的平台 白介素1β（IL-1β）的成熟和分泌。炎性小体的中心部分是 NLRP家族，该蛋白质与衔接蛋白ASC相关，进而报告促炎 caspase前体（例如procaspase-1）。在许多炎症中，NLRP3炎性体为 最广泛的研究。我们的工作假设是，接触金属纳米颗粒会导致 肺泡巨噬细胞（AMS）的炎性体和IL-1β分泌的激活和/或失调 肺上皮细胞和肺成纤维细胞，这将导致基质金属蛋白酶失调 （MMP）和金属蛋白酶（TIMP）的组织抑制剂，启动和促进金属纳米粒子 - 诱导肺损伤和纤维化。该项目将同时使用体外和体内系统来解决 以下特定目的：（1）确定炎性体激活在由IL-1β分泌中的作用 金属纳米颗粒在体外和体内。我们将确定是否涉及炎症的激活 在金属纳米颗粒诱导的IL-1β分泌中，肺泡巨噬细胞（AMS），肺上皮细胞和肺 成纤维细胞作者：（1）使用caspase-1的特定抑制剂AC-YVAD-CMK； （2）敲门 通过使用CRISPR/CAS9技术，炎性组组件，例如NLRP3，ASC或caspase-1； （3） 使用NLRP3或ASC敲除小鼠。然后，我们将确定NADPH氧化物和/或线粒体是否是否 依赖性ROS的产生和钾外排参与金属纳米颗粒诱导的炎性体 激活和IL-1β分泌。我们还将测量暴露于金属纳米颗粒的小鼠中的IL-1β分泌。 （2） 检查IL-1β在MMPS和TIMPS表达改变和活性中的作用 暴露于金属纳米颗粒。虽然IL-1β是MMP-2和MMP-9活性的诱导的，但尚不清楚它如何 通过暴露于金属纳米颗粒的情况下调节MMP和TIMP。测试炎性体和IL-的作用 1β在调节MMP和TIMP中，AIM 1中的策略将用于抑制炎性体功能， （1）药物IL-1β抑制剂将采用抑制IL-1功能的策略； （2）反il- 1β抗体； （3）对IL-1β无反应的IL-1RI - / - 小鼠。暴露于金属纳米颗粒后，MMP-2， 将确定MMP-9和TIMP的表达和活动。 （3）研究炎性体的作用 金属纳米颗粒引起的肺损伤和体内纤维化的激活。我们将首先调查是否 接触金属纳米颗粒会导致肺成纤维细胞产生更多的胶原蛋白。然后 金属纳米颗粒引起的肺纤维化中的炎症体将通过短期和长期暴露探索 小鼠到金属纳米颗粒。我们将在AIM 1和2中使用这些策略来抑制炎症体和IL-1β功能 研究炎性体和IL-1β在金属纳米颗粒诱导的肺纤维化中的作用。