Nguyen Theresa V, Ukairo Okechukwu, Khetani Salman R, McVay Michael, Kanchagar Chitra, Seghezzi Wolfgang, Ayanoglu Gulesi, Irrechukwu Onyi, Evers Raymond
Merck Research Laboratories, Department of Pharmacokinetics, Pharmacodynamics, and Drug Metabolism, Rahway, New Jersey (T.V.N., R.E.); Hepregen Corporation, Medford, Massachusetts (O.U., O.I.); Colorado State University, Fort Collins, Colorado (S.R.K.); Agios Pharmaceuticals, Cambridge, Massachusetts (M.M.); University Of Massachusetts Medical School, Department of Molecular Medicine, Worcester, Massachusetts (C.K); and Merck Research Laboratories, Department of Bioanalytics, Palo Alto, California (W.S., G.A)
Merck Research Laboratories, Department of Pharmacokinetics, Pharmacodynamics, and Drug Metabolism, Rahway, New Jersey (T.V.N., R.E.); Hepregen Corporation, Medford, Massachusetts (O.U., O.I.); Colorado State University, Fort Collins, Colorado (S.R.K.); Agios Pharmaceuticals, Cambridge, Massachusetts (M.M.); University Of Massachusetts Medical School, Department of Molecular Medicine, Worcester, Massachusetts (C.K); and Merck Research Laboratories, Department of Bioanalytics, Palo Alto, California (W.S., G.A).
Drug Metab Dispos. 2015 May;43(5):774-85. doi: 10.1124/dmd.114.061317. Epub 2015 Mar 4.
Elevated levels of proinflammatory cytokines associated with infection and inflammation can modulate cytochrome P450 enzymes, leading to potential disease-drug interactions and altered small-molecule drug disposition. We established a human-derived hepatocyte-Kupffer cell (Hep:KC) coculture model to assess the indirect cytokine impact on hepatocytes through stimulation of KC-mediated cytokine release and compared this model with hepatocytes alone. Characterization of Hep:KC cocultures showed an inflammation response after treatment with lipopolysaccharide and interleukin (IL)-6 (indicated by secretion of various cytokines). Additionally, IL-6 exposure upregulated acute-phase proteins (C-reactive protein, alpha-1-acid glycoprotein, and serum amyloid A2) and downregulated CYP3A4. Compared with hepatocytes alone, Hep:KC cocultures showed enhanced IL-1β-mediated effects but less impact from both IL-2 and IL-23. Hep:KC cocultures treated with IL-1β exhibited a higher release of proinflammatory cytokines, an increased upregulation of acute-phase proteins, and a larger extent of metabolic enzyme and transporter suppression. IC50 values for IL-1β-mediated CYP3A4 suppression were lower in Hep:KC cocultures (98.0-144 pg/ml) compared with hepatocytes alone (IC50 > 5000 pg/ml). Cytochrome suppression was preventable by blocking IL-1β interaction with IL-1R1 using an antagonist cytokine or an anti-IL-1β antibody. Unlike IL-1β, IL-6-mediated effects were comparable between hepatocyte monocultures and Hep:KC cocultures. IL-2 and IL-23 caused a negligible inflammation response and a minimal inhibition of CYP3A4. In both hepatocyte monocultures and Hep:KC cocultures, IL-2RB and IL-23R were undetectable, whereas IL-6R and IL-1R1 levels were higher in Hep:KC cocultures. In summary, compared with hepatocyte monocultures, the Hep:KC coculture system is a more robust in vitro model for studying the impact of proinflammatory cytokines on metabolic enzymes.
与感染和炎症相关的促炎细胞因子水平升高可调节细胞色素P450酶，导致潜在的疾病-药物相互作用以及小分子药物处置的改变。我们建立了一种人源肝细胞-库普弗细胞（Hep:KC）共培养模型，以通过刺激KC介导的细胞因子释放来评估细胞因子对肝细胞的间接影响，并将该模型与单独的肝细胞进行比较。Hep:KC共培养物的特征表明，在用脂多糖和白细胞介素（IL）-6处理后出现炎症反应（通过各种细胞因子的分泌来指示）。此外，暴露于IL-6会上调急性期蛋白（C反应蛋白、α-1-酸性糖蛋白和血清淀粉样蛋白A2）并下调CYP3A4。与单独的肝细胞相比，Hep:KC共培养物显示出增强的IL-1β介导的效应，但IL-2和IL-23的影响较小。用IL-1β处理的Hep:KC共培养物表现出更高的促炎细胞因子释放、急性期蛋白上调增加以及代谢酶和转运体抑制程度更大。与单独的肝细胞（IC50>5000 pg/ml）相比，Hep:KC共培养物中IL-1β介导的CYP3A4抑制的IC50值更低（98.0-144 pg/ml）。使用拮抗剂细胞因子或抗IL-1β抗体阻断IL-1β与IL-1R1的相互作用可预防细胞色素抑制。与IL-1β不同，IL-6介导的效应在肝细胞单培养物和Hep:KC共培养物之间具有可比性。IL-2和IL-23引起的炎症反应可忽略不计，对CYP3A4的抑制作用最小。在肝细胞单培养物和Hep:KC共培养物中，均未检测到IL-2RB和IL-23R，而Hep:KC共培养物中IL-6R和IL-1R1水平更高。总之，与肝细胞单培养物相比，Hep:KC共培养系统是一种更强大的体外模型，用于研究促炎细胞因子对代谢酶的影响。
