Elevated Dietary Non-Esterified Fatty Acid Levels Impact BeWo Trophoblast Metabolism and Lipid Processing: A Multi-OMICS Outlook. Elevated Dietary Non-Esterified Fatty Acid Levels Impact BeWo Trophoblast Metabolism and Lipid Processing: A Multi-OMICS Outlook

Maternal obesity and gestational diabetes mellitus (GDM) are associated with adverse intrauterine environments that are high in circulating nutrients. These adverse environments facilitate impairments in placental mitochondrial function and nutrient processing that ultimately leads to an increased risk of the exposed offspring developing non-communicable cardiometabolic health disorders (e.g. obesity, type 2 diabetes, metabolic syndrome). Analysis of fasting serum lipid levels in the third trimester has highlighted that Palmitate (PA) and oleate (OA) are the most abundance non-esterified fatty acids (NEFAs) in circulation. More importantly, these important NEFAs are elevated in both obese and GDM pregnancies. As PA and OA are the most abundant NEFAs in the typical “Westernized Diet” they elevated levels in obese and GDM pregnancies may highlight increased maternal consumption of these fats and could be a link between poor diet and abberant placental function. However, the independent and combined impacts of dietary PA and OA on mitochondrial function and nutrient processing in placental trophoblast cells remains poorly defined. Thus, in this study we sought to utilize a multi-omics approach (combining transcriptomics with untargeted metabolomics and lipidomics) to elucidate the underlying impacts of elevated NEFA levels on BeWo placental trophoblast cells. We highlighted a significant enrichment in the Wikipathways Fatty Acid Biosynthesis Pathway in BeWo cytotrophoblast cells treated with a combination of PA and OA (P/O; 50 µM each). Further, we identified differential expression of genes involved in lipid processing (e.g. ACADVL, ACACB, ACSL5, CREB3L3, PLIN2, and SCD) in BeWo cytotrophoblast cells treated with dietary NEFA species. Overall, these data highlighted that alterations in lipid processing, storage, and oxidation are implicated in the development of placental dysfunction in response to elevated lipid levels. Overall design: 20 total BeWo cytotrophoblast samples cultured with dietary fats conjugated to BSA for 72 hours, 5 treated with F12K media supplemented with 0.3% Fatty Acid-Free BSA (BSA control); 5 treated with F12K media supplemented with 100 µM Palmitate (PA; 2:1 conjugation to BSA); 5 treated with F12K media supplemented with 100 µM Oleate (OA; 2:1 conjugation to BSA); 5 treated with F12K media supplemented with 50 µM each PA and OA (P/O; 2:1 conjugation to BSA)