Journal Article

Activation of the Nrf2 response by intrinsic hepatotoxic drugs correlates with suppression of NF-κB activation and sensitizes toward TNFα-induced cytotoxicity

de Bont, Hans, Vrieling, Harry, Herpers, Bram, Fredriksson, Lisa, Hendriks, Giel, Wink, Steven, Di, Zi, van de Water, Bob

2016

Overview

Drug-induced liver injury (DILI) is an important problem both in the clinic and in the development of new safer medicines. Two pivotal adaptation and survival responses to adverse drug reactions are oxidative stress and cytokine signaling based on the activation of the transcription factors Nrf2 and NF-κB, respectively. Here, we systematically investigated Nrf2 and NF-κB signaling upon DILI-related drug exposure. Transcriptomics analyses of 90 DILI compounds in primary human hepatocytes revealed that a strong Nrf2 activation is associated with a suppression of endogenous NF-κB activity. These responses were translated into quantitative high-content live-cell imaging of induction of a selective Nrf2 target, GFP-tagged Srxn1, and the altered nuclear translocation dynamics of a subunit of NF-κB, GFP-tagged p65, upon TNFR signaling induced by TNFα using HepG2 cells. Strong activation of GFP-Srxn1 expression by DILI compounds typically correlated with suppression of NF-κB nuclear translocation, yet reversely, activation of NF-κB by TNFα did not affect the Nrf2 response. DILI compounds that provided strong Nrf2 activation, including diclofenac, carbamazepine and ketoconazole, sensitized toward TNFα-mediated cytotoxicity. This was related to an adaptive primary protective response of Nrf2, since loss of Nrf2 enhanced this cytotoxic synergy with TNFα, while KEAP1 downregulation was cytoprotective. These data indicate that both Nrf2 and NF-κB signaling may be pivotal in the regulation of DILI. We propose that the NF-κB-inhibiting effects that coincide with a strong Nrf2 stress response likely sensitize liver cells to pro-apoptotic signaling cascades induced by intrinsic cytotoxic pro-inflammatory cytokines.

• Publisher: Springer Berlin Heidelberg
• Subject: Active Transport, Cell Nucleus / Biomedical and Life Sciences / Biomedicine / Chemical and Drug Induced Liver Injury - genetics / Chemical and Drug Induced Liver Injury - metabolism / Chemical and Drug Induced Liver Injury - pathology / Computational Biology / Databases, Genetic / Dose-Response Relationship, Drug / Environmental Health / Gene Expression Profiling - methods / Gene Expression Regulation - drug effects / Green Fluorescent Proteins - biosynthesis / Green Fluorescent Proteins - genetics / Hep G2 Cells / Hepatocytes - drug effects / Hepatocytes - metabolism / Hepatocytes - pathology / Humans / Liver - drug effects / Liver - metabolism / Liver - pathology / NF-E2-Related Factor 2 - genetics / NF-E2-Related Factor 2 - metabolism / Occupational Medicine/Industrial Medicine / Organ Toxicity and Mechanisms / Oxidoreductases Acting on Sulfur Group Donors - biosynthesis / Oxidoreductases Acting on Sulfur Group Donors - genetics / Pharmacology/Toxicology / Recombinant Fusion Proteins - biosynthesis / RNA Interference / RNA, Messenger - genetics / RNA, Messenger - metabolism / Signal Transduction - drug effects / Time Factors / Transcription Factor RelA - genetics / Transcription Factor RelA - metabolism / Transfection / Tumor Necrosis Factor-alpha - toxicity