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

Abstract

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.

Toxicity of TiO2 Nanoparticles: Validation of Alternative Models

“…Consequently, the cells can be heterogeneously exposed with some cells exposed in excess and some not exposed at all [ 53 ]. High numbers of DEG genes between for in vitro submerged cells was already observed in previous transcriptomics studies [ 55 , 56 , 57 ]. This is why we chose to focus on DEGs with a FC > 2.8 for in vitro submerged exposition.…”

Systematic Review of Multi-Omics Approaches to Investigate Toxicological Effects in Macrophages

“…Doumandji et al, 2020 [ 136 ] studied the effects of ZnO and ZnFe 2 O 4 NMs in alveolar macrophages (NR8383) using 4 h exposure for transcriptomics and 24 h for proteomics. Importantly, proteomics was not only applied to cell extracts but also to cell culture supernatants.…”

“…The proteome combines both layers, being a mediator between environmental conditions and cellular processes [ 144 , 145 ]. While for the combination of transcriptomic and proteomic data high accordance of the detected biological processes has been described [ 134 , 135 , 136 , 137 , 138 ], combining the proteome with the metabolome, also referred to as proteometabolomics [ 146 , 147 , 148 , 149 , 150 ], has shown the advantage of complementing each other, thus allowing for a better understanding of triggered processes upon macrophage treatment with xenobiotics. Thus, based on the here presented publications, the combination of proteome and metabolome seems to be preferable even though both combinations might be generally beneficial to decipher xenobiotics MoAs.…”

Harmful effects of metal(loid) oxide nanoparticles

“…More specifically, it was found that a wide range of key metabolites involved in central carbon metabolism, cofactors synthesis, amino acid and fatty acid biosynthesis, purines and pyrimidines, nucleoside and nucleotide biosynthetic pathways were repressed (Babele 2019 ). By a similar approach (transcriptomic and proteome profile analysis), it was found that ZnO and ZnFe 2 O 4 NPs induced dysfunction of cholesterol biosynthesis in an alveolar rat macrophage cell line (Doumandji et al 2020 ).…”