GSE122151
Transcriptomic Analysis on Lipid Loaded HepaRG Model for Steatosis Reveal the Influence on the Regulation of Drug Metabolizing Enzymes and Transporters
The present study was aimed to mimic steatosis in in-vitro HepaRG model and test the effect of lipid loading on drug metabolizing enzymes and transporters (DMET) in steatosis.  HepaRG cells, differentiated in-house were exposed to mixture of saturated and unsaturated fatty acids, 0.5 mM palmitate and oleate (1:2) conjugated to BSA (0.02%) for 72h.  Following the incubation, the model system was characterized using ORO staining for neutral lipids, tested for toxicity with WST-1 assay and triglyceride (TAG) accumulation was measured using TAG calorimetric assay.  Increased lipid accumulation and 6-fold increase in TAG was observed (P < 0.05, n=4).  The whole transcriptome was analyzed using RNA sequencing.  Differentially expressed transcripts were identified upon FA treatment versus (vs.) control in HepaRG cell line (393 total transcripts, 89 up- and 304 downregulated).  Lipid loading, led to significant down-regulation in gene expression levels of various DMET, nuclear receptors and transcription factors such as CYP3A4, 1A2, 2B6, 2C8, 2C9 and 2C19, along with this UGT1A9, UGT1A6, ABCB1, NR1I2 and HNF4a (P < 0.05, n=3).  The accumulation of saturated and polyunsaturated fatty acids may selectively inhibit certain DME pathways. Overall design: Examination of drug metabolizing enzymes dyregulation in the presence of Oleate and Palmitate (2:1) enriched media in HepaRG cells