Multi-parametric profiling of a large panel of engineered nanomaterials (ENM) points to surface chemistry as a key determinant of nanomaterial effects
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PROVIDER: PRJNA625670 | ENA |
REPOSITORIES: ENA
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Project description:Here, we investigated a panel of 31 engineered nanomaterials (ENMs) with different core chemistries and surface modifications using conventional cytotoxicity assays coupled with omics-based approaches. Proteomics analysis were conducted in order to monitor changes in cells proteome exposed to ENMs. In this standard approach, cells are exposed to various types and amounts of ENMs and proteins are extracted after a given exposure period. For controls, no ENM are applied but the incubation and extraction of proteins is done the same way. Proteome changes are monitored using high-resolution Fourier transform mass spectrometry (Orbitraps) as well as in-house developed label-free software. Similar approaches have been used for quantitative analysis of posttranslational modifications. Proteomics analyses following low-dose exposure of THP-1 cells suggested a non-specific stress response to ENMs while microarray-based profiling revealed significant changes in gene expression as a function of both surface functionalization and core chemistry of the ENMs. Pathway analysis revealed that the most biologically active ENMs displaying cationic surfaces downregulated DNA replication/cell cycle responses and upregulated inflammatory responses. This study shows that surface chemistry is a key determinant of nanomaterial effects on immune cells.
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Project description:Proteomic data from engineered nanomaterial- treated THP-1 cells. Time points at 12 hours; 4 biological replicates for each treatment. Human THP-1 cells were either mock-treated (control) or treated with engineered nanomaterials (ENMs). Four biological replicates were used for each sample group (Control, or ENM treatment at either the low or high dosage). In the block design scheme for LC-MS analysis, samples treated with two different ENMs were assign into an analytical block (2 ENM X 2 dosages/ENM X 4 replicates = 16 samples). Four control samples were also included in each block, yielding 20 samples per block. Blocks of samples were divided into two processing batches. Samples in each batch were processed simultaneously from cell culture, to treatment with ENMs, and finally to proteomic sample preparation for LC-MS analysis (including trypsin digestion).
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Project description:This study aims to implement methods for toxicological profiling of engineered nanomaterials using toxicogenomics, proteomics, and metabolomics along with computational analyses. For all three omics layers the human cell lines A549 (lung epithelial cells) and THP1 (monocytes) were separately exposed to the nanomaterials TiO2 NM104, MWNCT NM401, and Ag NM300k. Proteomics and metabolomics samples have been performed as biological triplicates and were taken after 0, 6, 12, 24, and 48 hours. To assess ecotoxic effects C. elegans worms were grown in soil treated with NM300k. Ecotox samples were taken only at 0 and 24 hours. Integrating all three omics layers will enable the identification of (novel) ENM specific modes of action (MoA).
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Project description:The immune system may respond to engineered nanomaterials (ENM) through inflammatory reactions. The NLRP3 inflammasome responds to a wide range of ENM, and its activation is associated with various inflammatory diseases. The objective of the study was to compare the effects of gold ENM of different shapes on NLRP3 inflammasome activation and related signalling pathways. Differentiated THP-1 cells (wildtype, ASC- or NLRP3-deficient), were exposed to PEGylated gold nanorods, nanostars, and nanospheres. Exposed cells were subjected to gene expression analysis. Nanorods, but not nanostars or nanospheres, showed NLRP3 inflammasome activation. ASC- or NLRP3-deficient cells did not show this effect. Gold nanorod-induced NLRP3 inflammasome activation was accompanied by downregulated sterol/cholesterol biosynthesis, oxidative phosphorylation, and purinergic receptor signalling. In conclusion, the shape and surface chemistry of gold nanoparticles determine NLRP3 inflammasome activation.
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Project description:Due to the widespread application of food-relevant inorganic nanomaterials, the gastrointestinal tract is potentially exposed to these materials. Gut-on-chip in vitro model systems are proposed for the investigation of compound toxicity as they better recapitulate the in vivo human intestinal environment than static models, due to the added shear stresses associated with the flow of medium in line with what cells experience in vivo. We aimed to compare the cellular responses of intestinal epithelial Caco-2 cells at the gene expression level upon TiO2 (E171) and ZnO (NM110) nanomaterial exposure when cultured under dynamic and static conditions. For this, we applied whole genome transcriptome analyses. Differentially expressed genes and related biological processes revealed culture condition specific responses upon exposure to TiO2 and ZnO nanomaterials. The materials had more effects on cells cultured in the gut-on-chip when compared to the static model, indicating that shear stress might be a major factor in cell susceptibility. This is the first report on application of a gut-on-chip system to evaluate cellular responses upon TiO2 and ZnO nanomaterials compared to a static system and extends current knowledge on nanomaterial-cell interactions and toxicity assessment. Dynamically cultured cells appear to be more sensitive and the gut-on-chip might thus be an attractive model to be used more extensively in the toxicological hazard characterization.
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Project description:C57BL/6 mice were exposed by oropharyngeal aspiration to 28 nanomaterials including different surface functionalizations on 4 concecutive days with the dose of 10ug/day. Total RNA was collected from the mice lung biopsies after sacrificing.
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Project description:<p>This study aims to implement methods for toxicological profiling of engineered nanomaterials using toxicogenomics, proteomics, and metabolomics along with computational analyses. For all three omics layers the human cell lines A549 (lung epithelial cells) and THP1 (monocytes) were separately exposed to the nanomaterials TiO2 NM104, MWNCT NM401, and Ag NM300k. Proteomics and metabolomics samples have been performed as biological triplicates and were taken after 0, 6, 12, 24, and 48 hours. To assess ecotoxic effects C. elegans worms were grown in soil treated with NM300k. Ecotox samples were taken only at 0 and 24 hours. Integrating all three omics layers will enable the identification of (novel) ENM specific modes of action (MoA).</p><p><br></p><p><strong>Data availability:</strong></p><p>Proteomic data associated with this study are available in the PRIDE repository: accession number <a href='https://www.ebi.ac.uk/pride/archive/projects/PXD002401' rel='noopener noreferrer' target='_blank'>PXD002401</a>.</p>
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