Project 1: Mechanisms of Nanomaterial-Cell Interactions

项目1：纳米材料-细胞相互作用机制

基本信息

批准号：
8378137
负责人：
Brian Thrall
金额：
$ 55.38万
依托单位：
BATTELLE PACIFIC NORTHWEST LABORATORIES
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8378137
关键词：
Active Biological Transport Affect Air Pollution Alveolar Macrophages Artificial nanoparticles Attention Biodistribution Bioinformatics Biological Biological Assay Biometry Breathing Cell Communication Cell Wall Cell physiology Cells Charge Chemistry Clathrin Collaborations Data Dependence Detection Development Dose Environment Event Experimental Designs Exposure to Flow Cytometry Genes Genetic Genomics Goals Hospitalization Human Immune In Vitro Infection Inflammatory Kupffer Cells Ligands Liquid substance Lung Lysosomes Magnetism Measurement Measures Mediating Microscopy Modeling Molecular Mus National Institute of Environmental Health Sciences Natural Immunity Organelles Oxides Particulate Pathway interactions Pattern recognition receptor Phagocytosis Physiological Physiology Play Polystyrenes Predisposition Property Quantitative Structure-Activity Relationship Regulatory Pathway Research Research Personnel Resources Risk Risk Assessment Role SR-A proteins Science Serum Proteins Signal Transduction Silicon Dioxide Solubility Streptococcus pneumoniae Structure Surface System Testing base biodosimetry cell type cellular imaging ceric oxide clinically relevant design dosimetry exposed human population feeding hazard in vitro Model in vivo innate immune function intercellular communication iron oxide killings macrophage macrophage scavenger receptors member model development nanomaterials oxidized lipid particle pathogen receptor receptor expression receptor function response scavenger receptor trafficking uptake

项目摘要

The overarching goal of Project 1 is to understand how the physicochemical composition and structure of engineered nanoparticles (ENPs) dictate their biological interaction with cells to ultimately affect signaling and physiology. We hypothesize that scavenger receptor pathways plays a prominent role in determining the intracellular dose of ENPs in macrophages and biodistribution of ENPs in vivo. We further hypothesize that ENPs that "hijack' scavenger receptor uptake pathways may compromise macrophages ability to phagocytose and/or kill common bacterial lung pathogens. To test these hypotheses, microscopy, flow cytometry, and magnetic particle detection (MPD) will be used to quantitatively determine how varying the size, surface chemistry, charge and agglomeration state of ENPs (cerium oxides, silica oxides, iron oxides) affects their rate of uptake and trafficking pathways in macrophages derived from wildtype mice and mice deficient in class A scavenger receptors (Aim1). We will determine how the physicochemical properties of the ENPs affect stability of lysosomes, and their ability to activation inflammasome signaling (Aim 2). In vitro macrophage infection studies will be conducted to determine whether ENPs internalized by scavenger receptor pathways compromise the macrophages ability to recognize, phagocytosis and kill pathogens, using streptococcus pneumoniae as a model human lung pathogen (Aim 3). Finally, we will apply genomic microarray and bioinformatic analyses to identify gene regulatory pathways modulated by scavenger receptors and expore mechanisms by which ENPs may affect innate immune functions of macrophages (Aim 4). The dose-response studies in this project are designed to feed data directly for quantitative structure activity relationship analysis and development of dosimetry and risk models in Project 3. The results will also provide a molecular basis for experimental design and interpretation of parallel in vivo biodistribution and pathogen infection studies to be conducted in Project 2. We envision the proposed approach can provide quantitative data for hazard and risk analysis that is focused on clinically relevant measures of macrophage function that are potentially impacted by low level exposures to ENPs.

项目 1 的总体目标是了解工程纳米颗粒 (ENP) 的物理化学组成和结构如何决定它们与细胞的生物相互作用，从而最终影响信号传导和生理学。我们假设清道夫受体途径在决定巨噬细胞中 ENP 的细胞内剂量和 ENP 体内的生物分布。我们进一步假设“劫持”清道夫受体摄取途径的 ENP 可能会损害巨噬细胞吞噬和/或杀死常见细菌性肺部病原体的能力。为了检验这些假设，将使用显微镜、流式细胞术和磁粉检测 (MPD) 来定量确定 ENP（氧化铈、氧化硅、氧化铁）的尺寸、表面化学、电荷和团聚状态的变化如何影响其摄取和运输速率我们将确定来自野生型小鼠和缺乏 A 类清道夫受体 (Aim1) 的巨噬细胞的理化特性。 ENP 影响溶酶体的稳定性及其激活炎性体信号传导的能力（目标 2）。将进行体外巨噬细胞感染研究，以确定清道夫受体途径内化的 ENP 是否会损害巨噬细胞识别、吞噬和杀死病原体的能力，使用肺炎链球菌作为人类肺部病原体模型（目标 3）。最后，我们将应用基因组微阵列和生物信息学分析来识别清道夫受体调节的基因调控途径，并揭示ENP可能影响巨噬细胞先天免疫功能的机制（目标4）。该项目中的剂量反应研究旨在直接为项目 3 中的定量结构活性关系分析以及剂量测定和风险模型的开发提供数据。结果还将为实验设计和并行体内生物分布和风险模型的解释提供分子基础。项目 2 中将进行病原体感染研究。我们预计所提出的方法可以提供用于危害和风险分析的定量数据，重点关注可能受到低水平 ENP 暴露影响的巨噬细胞功能的临床相关测量。