Transcriptomic investigation of nanoparticle toxicity on HepaRG spheroids

Abstract

Nanoparticles (NPs) are used in diverse fields such as cosmetics, drug delivery, food industry and medicine. They are easy to produce and easy to modify according to the needs of the application. However, there is an arising concern about the adverse effects of the nanoparticles. Negative impacts of the nanoparticles on human body needs further analysis because nanoparticle exposure of human body increases every day with the widespread usage of these materials in daily products. In literature, the studies about the nanoparticle toxicity remain inadequate since they generally use insufficient number and non-variety of nanoparticles, inappropriate cell line or cell culture method. This study focuses on evaluation of nanoparticle toxicity with transcriptomic analysis. With the help of these analysis, we aim to find the genes that respond to NP toxicity, which might help us better understand the mechanism of the toxicity. HepaRG cells were used as the best alternative to primary hepatocytes. Additionally, to get the results most similar to the in vivo liver organ, HepaRG cells were cultured in 3D spheroids then they were incubated with eleven different nanoparticles of varying materials and sizes. To reveal the genes that responds to nanoparticle toxicity and to find the toxicity mechanisms of nanoparticles on HepaRG spheroids, transcriptome sequencing (RNA-seq) was conducted. Anticipatedly, cellular processes such as drug metabolism, sensitivity to metal ions, oxidative stress, EMT and apoptosis pathways showed enrichement in the DEGs of all nanoparticles. The common and unique genes inbetween different types and sizes of NPs were noted. These results offer novel findings about the toxicity mechanisms of HepaRG spheroids in response to NPs.