Biomimetic Microsystem for High Throughput Evaluation of Engineered Nanomaterials

用于工程纳米材料高通量评估的仿生微系统

• 批准号: 7941830
• 负责人: ROBERT M WORDEN
• 金额: $ 44.04万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-28 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7941830
• 关键词: Adjuvant; Affect; Allergens; Allergic; Allergic Disease; Animals; Antigens; Asthma; Behavior; Binding; Biological; Biological Assay; Biological Response Modifiers; Biomimetics; Breathing; CD8B1 gene; Cell Culture Techniques; Cell membrane; Cells; Charge; Chemistry; Computer Simulation; Data; Dendritic Cells; Deposition; Detection; Disease; Electrodes; Endocytosis; Engineering; Evaluation; Exhibits; Exposure to; Flow Cytometry; Fluorescence; Goals; Health; Health Status; Immune; Immune response; Immune system; In Vitro; Incidence; Inhalation Exposure; Interdisciplinary Study; Ligands; Lipid Bilayers; Lipopolysaccharides; Measurement; Measures; Membrane; Methods; Microelectrodes; Modeling; Modification; Molecular; Molecular Models; Mus; Ovalbumin; Oxidation-Reduction; Performance; Production; Property; Proteins; Quinones; Reaction; Reactive Oxygen Species; Risk Assessment; Role; Safety; Screening procedure; Severities; Spectrum Analysis; T cell response; T-Cell Activation; T-Lymphocyte; Testing; Theoretical model; Toxic effect; Toxicity Tests; allergic airway disease; allergic airway inflammation; base; chemical group; chemical property; commercial application; crosslink; design; electric impedance; engineering design; functional group; in vivo; insight; mathematical model; meetings; microsystems; molecular modeling; nanomaterials; nanoparticle; public health relevance; receptor; research study; response; uptake

DESCRIPTION (provided by applicant): Engineered nanomaterials (ENM) have unique properties that can cause adverse health effects. Due to their small size and potential for airborne dispersion, inhalation exposure to ENM might contribute to the increased incidence and/or exacerbation of allergic airway disease. The overall goal of this project is to develop a multi- level toxicity testing platform for ENM that includes in vivo measurement of allergic airway disease in mice, in vitro measurement of T cell activation, high-throughput measurement of ENM's interactions with bilayer lipid membranes (BLM) that mimic cell membranes, and in silico prediction of ENM's molecular properties. The overarching hypothesis that ENM possess adjuvant-like properties that promote allergic airway disease will be tested using five specific aims (SA). SA1 is to synthesize well-characterized ENM having controlled functional groups that catalyze redox reactions or activate membrane receptors. SA2 is to determine the adjuvant potential of ENM on allergic airway sensitization and asthma-like disease in mice. SA3 is to determine the effects of ENM on dendritic cell-induced activation and effector function of CD4+ and CD8+ T cells. SA4 is to measure the direct effects of ENM on synthetic bilayer lipid membranes. SA5 is to develop and validate mathematical models that can correlate ENM physicochemical properties with their biological and toxicological effects at the animal, cell, and membrane levels for health risk assessment. Biodegradable poly(propargyl glycolide) nanoparticles will be synthesized and coated with chemical groups (lipopolysaccharide (LPS) and quinone) to generate ENM likely to stimulate the immune response. LPS bind to receptor proteins on cell membranes and trigger cellular uptake by endocytosis, and quinones can trigger oxidation-reduction reactions, including production of reactive oxygen species by immune cells. A multi-tiered approach will be used to determine whether addition of LPS and quinone to ENM increases the ENM's ability to promote airway disease by (1) increasing the murine immune system's response to the antigen ovalbumin, (2) increasing T cell activation by dendritic cells in response to ovalbumin, and (3) modifying the ELM's molecular interactions with BLM. Mice will be exposed to the ENM by inhalation, and severity of allergic airway disease will be histopathologically, morphometrically and biochemically assessed. Dendritic cells will be exposed to the ENM, and their ability to activate T cells will be measured using fluorescence assisted flow cytometry. BLM will be deposited on electrodes and exposed to the ENM. The resulting interactions between the ENM and BLM will be measured in a high-throughput mode using cyclic voltammetry and electrical impedance spectroscopy. Theoretical models will be developed that describe the molecular properties of the ENM and their interactions with cellular components. These models will be used to analyze the experimental data and help elucidate mechanisms by which ENM induce toxic effects. PUBLIC HEALTH RELEVANCE: This project will provide fundamental insight into how a nanoparticle's physical and chemical properties determine its ability to enhance allergic airway disease like asthma. This insight will aid in setting health and safety standards for engineered nanomaterials, provide new high- throughput methods for nanoparticle detection and safety screening, and facilitate design of new nanomaterials that simultaneously meet safety standards and exhibit desirable performance properties needed for commercial applications.

描述（由申请人提供）：工程纳米材料（ENM）具有独特的特性，可能会对健康造成不利影响。由于 ENM 体积小且有可能在空气中扩散，吸入暴露于 ENM 可能会导致过敏性气道疾病的发病率增加和/或恶化。该项目的总体目标是开发ENM多级毒性测试平台，包括小鼠过敏性气道疾病的体内测量、T细胞活化的体外测量、ENM与双层脂膜相互作用的高通量测量。 BLM）模拟细胞膜，并通过计算机模拟预测 ENM 的分子特性。 ENM 具有促进过敏性气道疾病的佐剂样特性的总体假设将使用五个特定目标 (SA) 进行测试。 SA1用于合成具有受控官能团的特征良好的ENM，这些官能团可催化氧化还原反应或激活膜受体。 SA2旨在确定ENM对小鼠过敏性气道致敏和哮喘样疾病的辅助潜力。 SA3 旨在确定 ENM 对树突状细胞诱导的 CD4+ 和 CD8+ T 细胞的激活和效应功能的影响。 SA4 用于测量 ENM 对合成双层脂质膜的直接影响。 SA5 将开发和验证数学模型，将 ENM 理化特性与其在动物、细胞和膜水平上的生物和毒理学效应相关联，以进行健康风险评估。将合成可生物降解的聚（炔丙基乙交酯）纳米颗粒，并涂上化学基团（脂多糖（LPS）和醌），以生成可能刺激免疫反应的 ENM。 LPS 与细胞膜上的受体蛋白结合，并通过内吞作用触发细胞摄取，而醌可以触发氧化还原反应，包括免疫细胞产生活性氧。将使用多层方法来确定向 ENM 添加 LPS 和醌是否会通过以下方式增加 ENM 促进气道疾病的能力：(1) 增加小鼠免疫系统对抗原卵清蛋白的反应，(2) 增加树突状细胞的 T 细胞活化细胞响应卵清蛋白，(3) 改变 ELM 与 BLM 的分子相互作用。小鼠将通过吸入暴露于 ENM，并通过组织病理学、形态测量和生化评估过敏性气道疾病的严重程度。树突状细胞将暴露于 ENM，并使用荧光辅助流式细胞术测量它们激活 T 细胞的能力。 BLM 将沉积在电极上并暴露于 ENM。 ENM 和 BLM 之间产生的相互作用将使用循环伏安法和电阻抗谱以高通量模式进行测量。将开发理论模型来描述 ENM 的分子特性及其与细胞成分的相互作用。这些模型将用于分析实验数据并帮助阐明 ENM 诱发毒性作用的机制。 公共健康相关性：该项目将提供关于纳米颗粒的物理和化学特性如何决定其增强哮喘等过敏性气道疾病的能力的基本见解。这种见解将有助于制定工程纳米材料的健康和安全标准，为纳米颗粒检测和安全筛选提供新的高通量方法，并促进新纳米材料的设计，使其同时满足安全标准并表现出商业应用所需的理想性能。