The impact of SLC2A8 RNA interference on glucose uptake and the transcriptome of human trophoblast cells
While glucose is the primary fuel for fetal growth, the placenta utilizes the majority of glucose taken up from the maternal circulation. Of the facilitative glucose transporters in the placenta, SLC2A8 (GLUT8) is thought to primarily function as an intracellular glucose transporter, however its function in trophoblast cells has not been determined. To gain insight into the function of SLC2A8 in the placenta, lentiviral-mediated RNA interference (RNAi) was performed in the human first trimester trophoblast cell line ACH-3P. Non-targeting sequence controls (NTS RNAi; n=4) and SLC2A8 RNAi (n=4) infected ACH-3P cells were compared. A 79% reduction (P=0.06) in SLC2A8 mRNA concentration was associated with an 11% reduction (P=0.05) in ACH-3P glucose uptake. NTS RNAi and SLC2A8 RNAi ACH-3P mRNA was subjected to RNAseq, identifying 1525 transcripts that were differentially expressed (|log2FC|  > 1 and adjusted p-value < 0.05), with 273 transcripts derived from protein coding genes, and the change in 10 of these mRNA was validated by real-time qPCR. Additionally, there were 147 differentially expressed long non-coding RNAs. Functional analyses revealed differentially expressed genes involved in various metabolic pathways associated with cellular respiration, oxidative phosphorylation and ATP synthesis. Collectively, these data indicate that SLC2A8 deficiency may impact placental uptake of glucose, but that likely its primary function in trophoblast cells is to support cellular respiration. Since the placenta oxidizes the majority of the glucose it takes up to support its own metabolic needs, impairment of SLC2A8 function could set the stage for functional placental insufficiency.  This work was supported by National Institutes of Health grants HD094952 and HD093701.