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：
8069923
负责人：
Frank A Witzmann
金额：
$ 35.53万
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/8069923
关键词：
Agriculture Area Atomic Force Microscopy Attention Benign Bioinformatics Biological Biological Assay Carbon 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 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 Proteomics Reactive Oxygen Species Regulation Relative (related person)Research Research Personnel Residual state Resistance Resolution Risk Roentgen Rays Sampling Scanning Electron Microscopy Shapes Signal Transduction Skin Spectrometry Spectrum Analysis Spottings Surface Techniques Toxic effect Transmembrane Transport Up-Regulation Western Blotting Work X ray spectrometry base cell type cytokine density design in vivo injured interest irritation kidney cell monolayer multi walled carbon nanotube nanomaterials nanoparticle nanoscale particle programs protein expression protein function research study response single walled carbon nanotube statistics tandem mass spectrometry trafficking two-dimensional

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，制造的碳纳米颗粒（富勒烯、单壁纳米管和多壁纳米管）在暴露于屏障上皮（细支气管、肠和肾）细胞时，会针对膜和相关的细胞骨架蛋白发挥生物效应，从而改变细胞功能。具体来说，囊泡运输以及与囊泡运输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生电转运机制。我们认为这些效应与碳纳米颗粒的尺寸和物理化学特性直接相关，并且在某些特定剂量下可能对细胞造成伤害。因此，它们可能对健康造成重大危害。为了研究这个问题，我们计划暴露三个细胞在体外，使用跨越 3 个数量级的剂量响应设计，持续 24 和 48 小时，对上述三种碳纳米材料进行了类型分析。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技术来表征蛋白质翻译后修饰的变化。我们将通过测量离子传输和跨上皮电阻以及纳米颗粒暴露引起的对正常内源性调节剂反应的潜在变化来检查暴露的功能影响。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初级颗粒和团聚材料的元素组成、颗粒数量、粒度分布、颗粒形状和表面积，以及培养基团聚/解团聚状态。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整体蛋白表达的影响的重要新信息。此外，这些研究还将蛋白质表达的变化与毒性和亚毒性水平下细胞功能的变化联系起来。