The impact of hyperglycemia upon BeWo trophoblast cell metabolic function: A multi-OMICS and functional metabolic analysis

Abstract

Pre-existing and gestationally-developed diabetes mellitus have been linked with impairments in placental villous trophoblast cell metabolic function, that are thought to underlie the development of metabolic diseases early in the lives of the exposed offspring. Previous research using placental cell lines and ex vivo trophoblast preparations have highlighted hyperglycemia is an important independent regulator of placental function. However, it is poorly understood if hyperglycemia directly influences aspects of placental metabolic function, including nutrient storage and mitochondrial respiration, that are altered in term diabetic placentae. The current study examined metabolic and mitochondrial function as well as nutrient storage in both undifferentiated cytotrophoblast and differentiated syncytiotrophoblast BeWo cells cultured under hyperglycemia conditions (25 mM glucose) for 72 hours to further characterize the direct impacts of placental hyperglycemic exposure. Hyperglycemic-exposed BeWo trophoblasts displayed increased glycogen and triglyceride nutrient stores, but real-time functional readouts of metabolic enzyme activity and mitochondrial respiratory activity were not altered. However, specific investigation into mitochondrial dynamics highlighted increased expression of markers associated with mitochondrial fission that could indicate high glucose-exposed trophoblasts are transitioning towards mitochondrial dysfunction. To further characterize the impacts of independent hyperglycemia, the current study subsequently utilized a multi-omics approach and evaluated the transcriptomic and metabolomic signatures of BeWo cytotrophoblasts. BeWo cytotrophoblasts exposed to hyperglycemia displayed increased mRNA expression of ACSL1, HSD11B2, RPS6KA5, and LAP3 and reduced mRNA expression of CYP2F1, and HK2, concomitant with increased levels of: lactate, malonate, and riboflavin metabolites. These changes highlighted important underlying alterations to glucose, glutathione, fatty acid, and glucocorticoid metabolism in BeWo trophoblasts exposed to hyperglycemia. Overall, these results demonstrate that hyperglycemia is an important independent regulator of key areas of placental metabolism, nutrient storage, and mitochondrial function, and these data continue to expand our knowledge on mechanisms governing the development of placental dysfunction.

An Integrated Multi-OMICS Approach Highlights Elevated Non-Esterified Fatty Acids Impact BeWo Trophoblast Metabolism and Lipid Processing

“…Cellular metabolome profiles were analyzed using a High-Performance Chemical Isotope Labeling (HP CIL) liquid chromatography coupled mass spectrometry (LC-MS) approach, as previously described [ 67 , 72 , 73 ]. In brief, the 300 µL aliquot of supernatant was dried down and redissolved in water.…”

“…BeWo CT cells were plated at a density of 3.5 × 10 5 cells/well in 60 mm cell culture dishes and cultured with exogenous NEFAs, as described above. At T72H, the CT cells were washed once with PBS, collected in 0.9 mL TRIzol reagent (Invitrogen), and stored at −80 • C. Samples were then sent to the Genome Québec Innovation Centre for transcriptomic analysis using the Clariom S mRNA microarray (ThermoFisher Scientific, Mississauga, ON, Canada), as previously detailed [67]. In brief, RNA was automatically extracted using the QIAcube Connect (Qiagen, Toronto, ON, Canada).…”

Glucose, Insulin and Oxygen Modulate Expression of Serotonin-Regulating Genes in Human First-Trimester Trophoblast Cell Line ACH-3P

“…This approach aimed to provide more controlled and standardized experimental conditions by mitigating potential interference from insulin, glucose, and serotonin present in serum. The time points for treatments were selected based on published studies on the effects of glucose and insulin in trophoblast cellular models [ 27 , 28 ] and the results of our preliminary experiments.…”