Nanoparticle Effects on Epithelial Cell Protein Expression and Function

纳米颗粒对上皮细胞蛋白表达和功能的影响

基本信息

批准号：
7629090
负责人：
Frank A Witzmann
金额：
$ 36.25万
依托单位：
INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-06-01 至 2013-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7629090
关键词：
Agriculture Area Atomic Force Microscopy Attention Benign Bioinformatics Biological Biological Assay Carbon Carbon Nanotubes Carbon nanoparticle Cell Communication Cell Culture Techniques Cell Extracts Cell Survival Cell membrane Cell physiology Cells Characteristics Chemicals Computers Confocal Microscopy Confounding Factors (Epidemiology)Culture Media Cytoskeletal Proteins Data Dose Down-Regulation Electrical Resistance Electron Microscopy Electronics Electrophoresis Elements Environmental and Occupational Exposure Epithelial Epithelial Cells Epithelium Event Exposure to Extravasation Fluorescence Fullerenes Gel Goals Health Health Hazards Human IL8 gene Image Analysis In Vitro Incubated Individual Industry Inflammatory Injury Interleukin-10 Interleukin-13 Interleukin-6 Intestines Ion Transport Isoelectric Point Kidney Label Liquid Chromatography Lung Maintenance Manufacturer Name Maps Mass Spectrum Analysis Measurement Measures Mechanics Membrane Membrane Protein Traffic Membrane Proteins Metabolic Biotransformation Metals Methods Mining Modeling Modification Molecular Molecular Profiling Monitor Morphology Nanotechnology Nanotubes Occupational Organelles Particle Size Peptides Permeability Pharmacologic Substance Positioning Attribute Post-Translational Protein Processing Production Property Proteins Proteolysis Proteomics Reactive Oxygen Species Regulation Relative (related person)Research Research Personnel Residual state Resistance Resolution Risk Roentgen Rays Sampling Scanning Scanning Electron Microscopy Shapes Signal Transduction Skin Spectrometry Spectrum Analysis Spottings Surface Techniques Toxic effect Transmembrane Transport Western Blotting Work X ray spectrometry base cell type cytokine density design in vivo injured interest irritation kidney cell monolayer nanomaterials nanoparticle nanoscale particle programs protein expression research study response statistics tandem mass spectrometry trafficking two-dimensional

Agriculture Area Atomic Force Microscopy Attention Benign Bioinformatics Biological Biological Assay Carbon Carbon Nanotubes Carbon nanoparticle Cell Communication Cell Culture Techniques Cell Extracts Cell Survival Cell membrane Cell physiology Cells Characteristics Chemicals Computers Confocal Microscopy Confounding Factors (Epidemiology)Culture Media Cytoskeletal Proteins Data Dose Down-Regulation Electrical Resistance Electron Microscopy Electronics Electrophoresis Elements Environmental and Occupational Exposure Epithelial Epithelial Cells Epithelium Event Exposure to Extravasation Fluorescence Fullerenes Gel Goals Health Health Hazards Human IL8 gene Image Analysis In Vitro Incubated Individual Industry Inflammatory Injury Interleukin-10 Interleukin-13 Interleukin-6 Intestines Ion Transport Isoelectric Point Kidney Label Liquid Chromatography Lung Maintenance Manufacturer Name Maps Mass Spectrum Analysis Measurement Measures Mechanics Membrane Membrane Protein Traffic Membrane Proteins Metabolic Biotransformation Metals Methods Mining Modeling Modification Molecular Molecular Profiling Monitor Morphology Nanotechnology Nanotubes Occupational Organelles Particle Size Peptides Permeability Pharmacologic Substance Positioning Attribute Post-Translational Protein Processing Production Property Proteins Proteolysis Proteomics Reactive Oxygen Species Regulation Relative (related person)Research Research Personnel Residual state Resistance Resolution Risk Roentgen Rays Sampling Scanning Scanning Electron Microscopy Shapes Signal Transduction Skin Spectrometry Spectrum Analysis Spottings Surface Techniques Toxic effect Transmembrane Transport Western Blotting Work X ray spectrometry base cell type cytokine density design in vivo injured interest irritation kidney cell monolayer nanomaterials nanoparticle nanoscale particle programs protein expression research study response statistics tandem mass spectrometry trafficking two-dimensional

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项目摘要

DESCRIPTION (provided by applicant) Our hypothesis contends that manufactured carbon nanoparticles (fullerenes, single-walled nanotubes, and multi-walled nanotubes), when exposed to barrier epithelial (bronchiolar, intestinal, and kidney) cells, exert biological effects directed at the membrane and associated cytoskeletal proteins that alter cell function. Specifically, vesicular trafficking and the cytoskeletal components and events associated with vesicular trafficking are likely targets. Alterations in membrane-associated proteins will also alter or dysregulate electrogenic transport mechanisms. We propose that these effects are directly related to the size and physico-chemical character of the carbon nanoparticles, and, at some specific dose, are likely to be injurious to the cells. They thus may pose a significant health hazard. To study this, we plan to expose the three cell types to the three carbon nanomaterials mentioned above, in vitro, using a dose-response design spanning 3 orders of magnitude for a duration of 24 and 48 hrs. After exposure, we will examine differential protein expression using quantitative mass spectrometry and two-dimensional electrophoresis. We will also characterize alterations in protein post-translatiohal modification using those proteomic techniques. We will examine the functional effects of the exposures by measuring ion transport and transepithelial electrical resistance and the potential changes induced by nanoparticle exposure for the response to normal, endogenous regulators. In addition, we will quantify irritation/injury/toxicity using various measurements of cell cytokine secretion, reactive-oxygen species formation, and cell viability. An important aspect of this project is to accurately assess the elemental composition, particle number, particle size distribution, particle shape, and surface area of the primary particle and agglomerated material, as well as the culture-media agglomeration/de-agglomeration status. The results of these studies will provide important new information regarding the effect that different carbon nanoparticles have on barrier epithelial cell global protein expression. In addition, the studies will correlate the changes in protein expression with changes in cellular function at both toxic and subtoxic levels.

描述（由申请人提供）我们的假设认为，当暴露于膜上皮（支气管，肠道和肾脏）细胞时，生产碳纳米颗粒（富勒烯，单壁纳米管和多壁纳米管），对膜和相关的细胞骨骼蛋白质的生物学作用，这些效应的生物学作用。具体而言，囊泡贩运以及与水泡贩运相关的细胞骨架成分和事件可能是目标。膜相关蛋白的改变也会改变或失调的电源转运机制。我们建议这些作用与碳纳米颗粒的大小和物理化学特征直接相关，并且在某些特定剂量上可能对细胞有害。因此，它们可能构成重大的健康危害。为了研究这一点，我们计划暴露三个单元上面提到的三种碳纳米材料的类型在体外，使用剂量反应设计跨越3个数量级，持续时间为24和48小时。暴露后，我们将使用定量质谱和二维电泳检查差异蛋白表达。我们还将使用这些蛋白质组学技术来表征蛋白质后透透明修饰的改变。我们将通过测量离子转运和跨度电阻，以及纳米颗粒暴露引起的潜在变化来检查暴露的功能效应，以响应正常的内源性调节剂。此外，我们将使用细胞因子分泌，活性氧形成和细胞活力的各种测量来量化刺激/损伤/毒性。该项目的一个重要方面是准确评估主要粒子和团聚材料的元素组成，颗粒数，粒径分布，粒子形状和表面积，以及培养物媒体中的媒体团聚/脱水状态。这些研究的结果将提供有关不同碳纳米颗粒对屏障上皮细胞全局蛋白表达的影响的重要新信息。此外，研究将将蛋白质表达的变化与毒性和潜毒水平的细胞功能变化相关。