Fast and Robust Proteome Screening Platform Identifies Neutrophil Extracellular Trap Formation in the Lung in Response to Cobalt Ferrite Nanoparticles
Fast and Robust Proteome Screening Platform Identifies Neutrophil Extracellular Trap Formation in the Lung in Response to Cobalt Ferrite Nanoparticles
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DOI：
10.1021/acsnano.9b08818
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发表时间：
2020-04-28
期刊：
影响因子：
17.1
通讯作者：
Kjeldsen, Frank
中科院分区：
文献类型：
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作者：
Billing, Anja M.;Knudsen, Kristina B.;Kjeldsen, Frank
尽管磁性纳米粒子在生物医学和电子学中有广泛的应用，但只有少数关于生物相容性的体内研究。在这项研究中，磁性金属氧化物纳米颗粒对呼吸系统的毒性进行了检查，在体内通过单次气管内滴注小鼠。收集支气管肺泡灌洗液（BALF）样品，通过LC-MS/MS进行蛋白质组分析，在两个剂量（每只动物54 μ g，162 μ g）和两个时间点（滴注后第1天和第3天）测试掺杂有增加量的钴（Fe 3 O 4，CoFe 2 O 4，铁钴比为5：1，3：1，1：3，Co 3 O 4）的Fe 3 O 4纳米颗粒。在发现阶段，一些代表性样品的深入蛋白质组分析允许进行全面的途径分析。681个差异表达蛋白质的聚类（FDR < 0.05）揭示了一般以及金属氧化物特异性应答，具有先天免疫的总体强诱导和补体系统的活化。最高的表达增加可以被发现的一个集群的39种蛋白质，这显示出强烈的剂量依赖性氧化铁，可以归因于中性粒细胞胞外陷阱（NET）的形成。深入的蛋白质组分析扩展了体内NET形成的知识。在筛选期间，在使用Evosep One系统进行短梯度（21分钟）LC分离后，将研究的所有BALF样品（n = 166）作为单次注射进行无标记测量，从而验证来自发现的发现并定义能够区分肺部炎症的蛋白质特征。我们展示了一种基于蛋白质组学的毒性筛选，具有高样品通量，易于转移到其他纳米颗粒类型。数据可通过ProteomeXchange获得，标识符为PXDO 16148。
Despite broad application of magnetic nanoparticles in biomedicine and electronics, only a few in vivo studies on biocompatibility are available. In this study, toxicity of magnetic metal oxide nanoparticles on the respiratory system was examined in vivo by single intratracheal instillation in mice. Bronchoalveolar lavage fluid (BALF) samples were collected for proteome analyses by LC-MS/MS, testing Fe3O4 nanoparticles doped with increasing amounts of cobalt (Fe3O4, CoFe2O4 with an iron to cobalt ratio 5:1, 3:1, 1:3, Co3O4) at two doses (54 mu g, 162 mu g per animal) and two time points (day 1 and 3 days postinstillation). In discovery phase, in-depth proteome profiling of a few representative samples allowed for comprehensive pathway analyses. Clustering of the 681 differentially expressed proteins (FDR < 0.05) revealed general as well as metal oxide specific responses with an overall strong induction of innate immunity and activation of the complement system. The highest expression increase could be found for a cluster of 39 proteins, which displayed strong dose-dependency to iron oxide and can be attributed to neutrophil extracellular trap (NET) formation. In-depth proteome analysis expanded the knowledge of in vivo NET formation. During screening, all BALF samples of the study (n = 166) were measured label-free as single-injections after a short gradient (21 min) LC separation using the Evosep One system, validating the findings from the discovery and defining protein signatures which enable discrimination of lung inflammation. We demonstrate a proteomics-based toxicity screening with high sample throughput easily transferrable to other nanoparticle types. Data are available via ProteomeXchange with identifier PXDO16148.