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Fast and Robust Proteome Screening Platform Identifies Neutrophil Extracellular Trap Formation in the Lung in Response to Cobalt Ferrite Nanoparticles
Version 2 2020-03-30, 14:07
Version 1 2020-03-18, 20:15
journal contribution
posted on 2020-03-30, 14:07 authored by Anja M. Billing, Kristina B. Knudsen, Andrew J. Chetwynd, Laura-Jayne A. Ellis, Selina V. Y. Tang, Trine Berthing, Håkan Wallin, Iseult Lynch, Ulla Vogel, Frank KjeldsenDespite
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 μg, 162 μg 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 PXD016148.
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discovery phaseLC separationcomplement system162 μ glung inflammationcobaltintratracheal instillationproteomics-based toxicity screeningneutrophil extracellular trapFDRday 1protein signaturesCoFe 2 O 4BALF samplesrepresentative samplesBronchoalveolar lavage fluidformationmetal oxidenanoparticle typesexpression increasetime points681 differentially39 proteinsvivo studiespathway analysesFe 3 O 4metal oxide nanoparticlesRobust Proteome Screening Platform ...sample throughput3 days postinstillationPXDiron oxideproteome analysesIn-depth proteome analysisCobalt Ferrite Nanoparticles
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