Presenting Author Washington State University Spokane Valley, Washington
Acetaminophen (APAP) is the most used medication for pain and fever in pregnancy, but recent studies have elucidated in utero APAP as an endocrine disruptor associated with neurocognitive disorders. The placenta is a vital endocrine organ and disruption in metabolic pathways have been associated with adverse fetal neurodevelopment such as attention deficit hyperactivity disorder (ADHD) and autism. The aim of this study was to investigate the effect of APAP treatment on androgen pathways in JEG-3 placental cells.
JEG-3 cells were seeded in 6 well plates at 100,000 cells per well and treated with the vehicle control (DMSO) and 0.1, 0.3, and 0.9 mM APAP for 72 hours, in triplicate. Cells were harvested and samples were digested with trypsin. Global proteomics data were acquired using a Q-Exactive HF Quadrupole-Orbitrap mass spectrometer (Thermo Fisher Scientific, Waltham, MA) followed by data processing by MaxQuant (version 1.6.8.0) using the Andromeda search engine. Unique and significantly different (upregulated or downregulated) proteins were identified. Label-free quantification (LFQ) intensities of sex-steroid pathway proteins were compared across control and treatment groups.
Proteomics data analysis detected 2338 – 2560 total proteins with 1953 proteins in common between all treatments. The vehicle control group had 88 unique proteins and APAP treatment groups ranging from 0.1, 0.3, and 0.9 mM resulted 62, 55, and 157 unique proteins, respectively. Treatment with APAP upregulated ~200 proteins and downregulated ~40 proteins representing critical metabolic processes. APAP treatment elevated the level of AKR1B1 by at least 63% across all treatment groups. There was no change in the levels of SOAT1, CYP19A1, and STS in any treatment group. Interestingly, ABCG2 was uniquely detected in the lower APAP treatments.
Sex steroid metabolism plays a significant in utero role in brain and reproductive development of a fetus. Determining how APAP disrupts endocrine pathways is critical in managing pain treatment guidelines in pregnant women. This is the first study investigating the effect the APAP on sex steroid related proteome in placental cells, which when validated by our metabolic and activity assays, will provide a critical mechanistic information towards mitigating adverse effects of APAP.
