A combined proteomic and targeted analysis unravels new toxic mechanisms for zinc oxide nanoparticles in macrophages

Aude-Garcia, Catherine; Dalzon, Bastien; Ravanat, Jean‐Luc; Collin-Faure, Véronique; Diemer, Hélène; Strub, Jean‐Marc; Cianférani, Sarah; Dorsselaer, Alain Van; Carrière, Marie; Rabilloud, Thierry

doi:10.1016/j.jprot.2015.12.013

Cited by 45 publications

(52 citation statements)

References 84 publications

(132 reference statements)

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“…7 The main strongly deregulated proteins in NR8383 cells exposed to both ¼ IC 50 doses of ZnO NPs (a) and ZnFe 2 O 4 NPs (b) during 24 h. Proteins ratio (a): 16 × 10 6 ≤ R ZnO ≤ 0.18; protein ratio (b) 6 × 10 6 ≤ R ZnFe2O4 ≤ 2.6 × 10 −7 phosphorylation, and sirtuin pathways. Our data are in agreement with previous studies (Aude-Garcia et al 2016;Chevallet et al 2016;George and Ahmad 2016;Niska et al 2015). Altogether, these findings suggest that the regulation of these pathways might help cells capture and fix Zn 2+ and reduce the oxidative effect induced by ZnO and ZnFe 2 O 4 NPs.…”

Section: Discussionsupporting

confidence: 93%

Protein and lipid homeostasis altered in rat macrophages after exposure to metallic oxide nanoparticles

et al. 2019

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Metal oxide nanoparticles (NPs), such as ZnO, ZnFe 2 O 4 , and Fe 2 O 3 , are widely used in industry. However, little is known about the cellular pathways involved in their potential toxicity. Here, we particularly investigated the key molecular pathways that are switched on after exposure to sub-toxic doses of ZnO, ZnFe 2 O 4 , and Fe 2 O 3 in the in vitro rat alveolar macrophages (NR8383). As in our model, the calculated IC 50 were respectively 16, 68, and more than 200 μg/mL for ZnO, ZnFe 2 O 4 , and Fe 2 O 3 ; global gene and protein expression profiles were only analyzed after exposure to ZnO and ZnFe 2 O 4 NPs. Using a rat genome microarray technology, we found that 985 and 1209 genes were significantly differentially expressed in NR8383 upon 4 h exposure to ¼ IC 50 of ZnO and ZnFe 2 O 4 NPs, respectively. It is noteworthy that metallothioneins were overexpressed genes following exposure to both NPs. Moreover, Ingenuity Pathway Analysis revealed that the top canonical pathway disturbed in NR8383 exposed to ZnO and ZnFe 2 O 4 NPs was eIF2 signaling involved in protein homeostasis. Quantitative mass spectrometry approach performed from both NR8383 cell extracts and culture supernatant indicated that 348 and 795 proteins were differentially expressed upon 24 h exposure to ¼ IC 50 of ZnO and ZnFe 2 O 4 NPs, respectively. Bioinformatics analysis revealed that the top canonical pathways disturbed in NR8383 were involved in protein homeostasis and cholesterol biosynthesis for both exposure conditions. While VEGF signaling was specific to ZnO exposure, iron homeostasis signaling pathway was specific to ZnFe 2 O 4 NPs. Overall, the study provides resource of transcriptional and proteomic markers of response to ZnO and ZnFe 2 O 4 NP-induced toxicity through combined transcriptomics, proteomics, and bioinformatics approaches.

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Section: Discussionsupporting

confidence: 93%

Protein and lipid homeostasis altered in rat macrophages after exposure to metallic oxide nanoparticles

et al. 2019

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“…In conclusion, the first lesson from this work is that a simple medium switch induces more changes in the core cellular proteome than responses to low toxicity nanoparticles such as titanium dioxide or zinc dioxide . Somewhat surprisingly, the classes of proteins modulated by the medium change go much beyond the metabolism‐associated proteins in a wide sense (e.g.…”

Section: Discussionmentioning

confidence: 87%

“…This experiment was performed essentially as described previously . Cells were cultured on coverslips placed in 6‐well plates and exposed to NPs for 24 h at 37°C.…”

Section: Methodsmentioning

confidence: 99%

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Culture medium associated changes in the core proteome of macrophages and in their responses to copper oxide nanoparticles

et al. 2016

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The physiology of cells cultured in vitro depends obviously on the external conditions, including the nutrients present in the culture medium. In order to test the influence of this parameter, J774 macrophages grown either in RPMI or in DMEM were compared by a combination of targeted analyses and a proteomic approach. The two media differ in their glucose, amino acids, and vitamins concentrations, but there were no significant differences in the cell cycle or in the percentage of phagocytic cells in both media, although the phagocytic capacity (i.e. the number of phagocytized particles) was higher in DMEM. Conversely, we found that J774 cells grown in RPMI produced more nitric oxide in response to lipopolysaccharide. The proteomic study highlighted differences affecting the central metabolism and nucleotide metabolism, cytoskeleton, protein degradation, and cell signaling. Furthermore, proteomics showed that J774 cells grown in RPMI or in DMEM and exposed to copper oxide nanoparticles respond rather differently, with only a few proteins similarly modulated between cells grown in both media. Taken together, our results show that the basal state of cells grown in two different media is different, and this may affect the way they respond to an external stimulus or stress.

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“…The zinc ion has a strong affinity to sulfur and binds to glutathione (34) and to cysteine residues in protein active sites (35,36). This can result in the inhibition of key metabolic enzymes, ranging from glyceraldehyde phosphate dehydrogenase to mitochondrial enzymes (37)(38)(39). Interestingly, zinc toxicity is at least partly reversed by supplementation with metabolic end-products such as pyruvate and/or oxaloacetate (40)(41)(42)(43)(44)(45).…”

Section: Introductionmentioning

confidence: 99%

A Combined N-terminomics and Shotgun Proteomics Approach to Investigate the Responses of Human Cells to Rapamycin and Zinc at the Mitochondrial Level

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Aude-Garcia

et al. 2019

Molecular & Cellular Proteomics

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All but thirteen mammalian mitochondrial proteins are encoded by the nuclear genome, translated in the cytosol and then imported into the mitochondria. For a significant proportion of the mitochondrial proteins, import is coupled with the cleavage of a presequence called the transit peptide, and the formation of a new N-terminus. Determination of the neo N-termini has been investigated by proteomic approaches in several systems, but generally in a static way in order to compile as many N-termini as possible. In the present study, we have investigated how the mitochondrial proteome and N-terminome react to chemical stimuli that alter mitochondrial metabolism, namely zinc ions and rapamycin. To this end, we have used a strategy that analyzes both internal and N-terminal peptides in a single run, the dN-TOP approach. We used these two very different stressors to sort out what could be a generic response to stress and what is specific to each of these stressors. Rapamycin and zinc induced fairly different changes in the mitochondrial proteome. However, convergent changes to key mitochondrial enzymatic activities such as pyruvate dehydrogenase, succinate dehydrogenase and citrate synthase were observed for both treatments. Other convergent changes were seen in components of the N-terminal processing system and mitochondrial proteases. Investigations into the generation of neo N-termini in mitochondria showed that the processing system is robust, as indicated by the lack of change in neo N-termini under the conditions tested. Detailed analysis of the data revealed that zinc caused a slight reduction in the efficiency of the N-terminal trimming system and that both treatments increased the degradation of mitochondrial proteins. In conclusion, the use of this combined strategy allowed a detailed analysis of the dynamics of the mitochondrial N-terminome in response to treatments which impact the mitochondria.

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A combined proteomic and targeted analysis unravels new toxic mechanisms for zinc oxide nanoparticles in macrophages

Cited by 45 publications

References 84 publications

Protein and lipid homeostasis altered in rat macrophages after exposure to metallic oxide nanoparticles

Protein and lipid homeostasis altered in rat macrophages after exposure to metallic oxide nanoparticles

Culture medium associated changes in the core proteome of macrophages and in their responses to copper oxide nanoparticles

A Combined N-terminomics and Shotgun Proteomics Approach to Investigate the Responses of Human Cells to Rapamycin and Zinc at the Mitochondrial Level

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