Effect of Pro-inflammatory Cytokines on the mRNA Abundance of Major Intestinal Drug Transporters

Abstract ID 92080 Poster Board 032 In vivo studies suggest that inflammation or infections can markedly alter drug clearance, thereby affecting drug effectiveness and safety (Morgan E. T. et al., 2009). This alteration in drug clearance is caused by the elevation in plasma pro-inflammatory cytokines, which can modulate the in vitro expression and activity of human hepatic CYP enzymes and transporters as well as renal transporters. However, the impact of these cytokines on the human intestine, a critical site for drug absorption, remains underexplored, presenting challenges in applying these findings in vivo. This study aimed to systematically investigate the influence of pro-inflammatory cytokines on the mRNA abundance of major intestinal drug transporters using an in vitro human enteroid model. These cells, chosen for their robust drug transport and metabolizing activities, closely mimic the functions of the intestinal mucosal epithelial barrier. Enteroid monolayers derived from three duodenal crypt stem cell donors were cultured in 96-well format, differentiated for 14 days, and then exposed for 48 hours, in triplicate, to interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and interferon-γ (IFN-γ), either individually or in combination, at concentrations of 0.1, 1, and 10 ng/mL. These concentrations span from basal to supra-pathophysiological levels, corresponding to those observed in various infectious/inflammatory diseases. Following treatment, total RNA was isolated and subjected to quantitative PCR (qPCR) measurement of key intestinal transporter mRNAs: P-Glycoprotein (P-gp), Breast Cancer Resistance Protein (BCRP), Multidrug Resistance-Associated Protein 1, 2, 3, and 4 (MRP1/2/3/4), Organic Solute Transporter Alpha and Beta (OST-α/β), Organic Cation Transporter 1 and 3 (OCT1/3), Plasma Membrane Monoamine Transporter (PMAT), and Organic Anion Transporting Polypeptide 2B1 (OATP2B1). Exposure to a high concentration of pro-inflammatory cytokines down-regulated intestinal P-gp, PCRP, MRP2/3, OST α/ β, PMAT, and OATP2B1 mRNA levels, and these changes are expected to be concentration-dependent based on prior investigations of DME perturbations by cytokine exposure. To our knowledge, this is the first systematic investigation of the impact of cytokines on human intestinal drug transporters. Future work will focus on determining whether these changes in enteroid mRNA abundance correspond to alterations in intestinal transporter activity and/or protein abundance. These data are pivotal for developing physiologically based pharmacokinetic models to predict transporter-mediated drug pharmacokinetics during inflammation or infection. This work was supported by the NIH Grant R01HD102786.