Using Transcriptomic Signatures to Elucidate Individual and Mixture Effects of Inorganic Arsenic and Manganese in Human Placental Trophoblast HTR-8/SVneo Cells.

Abstract

Prenatal exposure to the toxic metal inorganic arsenic (iAs) is associated with adverse pregnancy and fetal growth outcomes. These adverse outcomes are tied to physiological disruptions in the placenta. While iAs co-occurs in the environment with other metals such as manganese (Mn), there is a gap in the knowledge of the effects of metal-mixtures on the placenta. To address this, we exposed human placental trophoblast cells to iAs, Mn, and an iAs-Mn mixture at three concentrations and evaluated transcriptome-wide gene expression and placental migration. We hypothesized that co-exposure to iAs-Mn in a mixture would result in a synergistic/enhanced transcriptomic effect compared to either metal alone. We also anticipated that genes involved in inflammatory or immune-related pathways would be differentially expressed in relation to the mixture compared to single-metals. The results highlight that iAs exposure alone had a stronger genomic response than Mn exposure, with two-fold the number of differentially expressed genes (DEGs). When analyzing DEGs present across all concentrations of study, the iAs-Mn mixture resulted in the greatest number of DEGs. The results highlight that iAs exposure alone influences the expression of toll-like receptor-initiated response pathways including Triggering Receptor Expressed on Myeloid Cells-1 TREM. Exposure to Mn alone influenced the expression of Nicotinamide adenine dinucleotide (NAD) biosynthesis pathways. In contrast, exposure to the iAs-Mn mixtures resulted in altered expression of inflammatory and immune response-related pathways, including the Nuclear factor erythroid 2 p45-related factor 2 (NRF2)-mediated oxidative stress response pathway. Migration was unaffected by iAs, Mn or the iAs-Mn mixture. These findings provide novel toxicogenomic insights into iAs and Mn-induced placental transcriptomic dysregulations at environmentally-relevant concentrations, with implications that in utero exposure to metal mixtures can influence inflammatory and immune pathways within the placenta.