Hepatic DNA deposition drives drug-induced liver injury and inflammation in mice

Authors

  • Pedro Elias Marques,

    1. Laboratório de Imunobiofotônica, Departamento de Morfologia, ICB, Universidade Federal de Minas Gerais, Minas Gerais, Brazil
    Search for more papers by this author
    • These authors contributed equally to this work.

  • André Gustavo Oliveira,

    1. Laboratório de Imunobiofotônica, Departamento de Morfologia, ICB, Universidade Federal de Minas Gerais, Minas Gerais, Brazil
    2. Departamento de Fisiologia e Biofísica, ICB, Universidade Federal de Minas Gerais, Minas Gerais, Brazil
    Search for more papers by this author
    • These authors contributed equally to this work.

  • Rafaela Vaz Pereira,

    1. Laboratório de Imunobiofotônica, Departamento de Morfologia, ICB, Universidade Federal de Minas Gerais, Minas Gerais, Brazil
    Search for more papers by this author
  • Bruna Araújo David,

    1. Laboratório de Imunobiofotônica, Departamento de Morfologia, ICB, Universidade Federal de Minas Gerais, Minas Gerais, Brazil
    Search for more papers by this author
  • Lindisley Ferreira Gomides,

    1. Laboratório de Imunobiofotônica, Departamento de Morfologia, ICB, Universidade Federal de Minas Gerais, Minas Gerais, Brazil
    Search for more papers by this author
  • Adriana Machado Saraiva,

    1. Laboratório de Imunobiofotônica, Departamento de Morfologia, ICB, Universidade Federal de Minas Gerais, Minas Gerais, Brazil
    Search for more papers by this author
  • Daniele Araújo Pires,

    1. Laboratório de Imunobiofotônica, Departamento de Morfologia, ICB, Universidade Federal de Minas Gerais, Minas Gerais, Brazil
    Search for more papers by this author
  • Júlia Tosta Novaes,

    1. Laboratório de Imunobiofotônica, Departamento de Morfologia, ICB, Universidade Federal de Minas Gerais, Minas Gerais, Brazil
    Search for more papers by this author
  • Daniel O. Patricio,

    1. Departamento de Microbiologia, Imunologia e Parasitologia, CCB, Universidade Federal de Santa Catarina, Santa Catarina, Brazil
    Search for more papers by this author
  • Daniel Cisalpino,

    1. Departamento de Microbiologia, ICB, Universidade Federal de Minas Gerais, Minas Gerais, Brazil
    Search for more papers by this author
  • Zélia Menezes-Garcia,

    1. Departamento de Microbiologia, Imunologia e Parasitologia, CCB, Universidade Federal de Santa Catarina, Santa Catarina, Brazil
    Search for more papers by this author
  • W. Matthew Leevy,

    1. Biological Imaging Core, University of Notre Dame, Department of Biological Sciences, South Bend, IN, USA
    Search for more papers by this author
  • Sarah Ellen Chapman,

    1. Biological Imaging Core, University of Notre Dame, Department of Biological Sciences, South Bend, IN, USA
    Search for more papers by this author
  • GermánArturo Mahecha,

    1. Departamento de Morfologia, ICB, Universidade Federal de Minas Gerais, Minas Gerais, Brazil
    Search for more papers by this author
  • Rafael Elias Marques,

    1. Laboratório de Imunofarmacologia, Departamento de Bioquímica e Imunologia, ICB, Universidade Federal de Minas Gerais, Minas Gerais, Brazil
    Search for more papers by this author
  • Rodrigo Guabiraba,

    1. INRA, Pôle Santé Animale de Tours PSAT, UMR 1282, Infectiologie et Santé Publique, Nouzilly, France
    Search for more papers by this author
  • Vicente Paulo Martins,

    1. Departamento de Biologia Celular (CEL), Instituto de Ciências Biológicas (IB), Universidade de Brasília (UnB), Brasília, DF, Brazil
    Search for more papers by this author
  • Danielle Gloria Souza,

    1. Departamento de Microbiologia, ICB, Universidade Federal de Minas Gerais, Minas Gerais, Brazil
    Search for more papers by this author
  • Daniel Santos Mansur,

    1. Departamento de Microbiologia, Imunologia e Parasitologia, CCB, Universidade Federal de Santa Catarina, Santa Catarina, Brazil
    Search for more papers by this author
  • Mauro Martins Teixeira,

    1. INRA, Pôle Santé Animale de Tours PSAT, UMR 1282, Infectiologie et Santé Publique, Nouzilly, France
    Search for more papers by this author
  • M. Fatima Leite,

    1. Departamento de Fisiologia e Biofísica, ICB, Universidade Federal de Minas Gerais, Minas Gerais, Brazil
    Search for more papers by this author
  • Gustavo Batista Menezes

    Corresponding author
    1. Laboratório de Imunobiofotônica, Departamento de Morfologia, ICB, Universidade Federal de Minas Gerais, Minas Gerais, Brazil
    • Address reprint requests to: Gustavo Batista Menezes, Ph.D., Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Antônio Carlos 6627, Pampulha, Belo Horizonte 31270-901, Minas Gerais, Brazil. E-mail: menezesgb@ufmg.br

    Search for more papers by this author

  • Potential conflict of interest: Nothing to report.

  • Supported by grants from CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico), FAPEMIG (Fundação de Amparo à Pesquisa de Minas Gerais), PRONEX, CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior) and Howard Hughes Medical Institute (HHMI).

  • See Editorial on Page 35

Abstract

Drug-induced liver injury (DILI) is an important cause of acute liver failure, with limited therapeutic options. During DILI, oncotic necrosis with concomitant release and recognition of intracellular content amplifies liver inflammation and injury. Among these molecules, self-DNA has been widely shown to trigger inflammatory and autoimmune diseases; however, whether DNA released from damaged hepatocytes accumulates into necrotic liver and the impact of its recognition by the immune system remains elusive. Here we show that treatment with two different hepatotoxic compounds (acetaminophen and thioacetamide) caused DNA release into the hepatocyte cytoplasm, which occurred in parallel with cell death in vitro. Administration of these compounds in vivo caused massive DNA deposition within liver necrotic areas, together with an intravascular DNA coating. Using confocal intravital microscopy, we revealed that liver injury due to acetaminophen overdose led to a directional migration of neutrophils to DNA-rich areas, where they exhibit an active patrolling behavior. DNA removal by intravenous DNASE1 injection or ablation of Toll-like receptor 9 (TLR9)-mediated sensing significantly reduced systemic inflammation, liver neutrophil recruitment, and hepatotoxicity. Analysis of liver leukocytes by flow cytometry revealed that emigrated neutrophils up-regulated TLR9 expression during acetaminophen-mediated necrosis, and these cells sensed and reacted to extracellular DNA by activating the TLR9/NF-κB pathway. Likewise, adoptive transfer of wild-type neutrophils to TLR9−/− mice reversed the hepatoprotective phenotype otherwise observed in TLR9 absence. Conclusion: Hepatic DNA accumulation is a novel feature of DILI pathogenesis. Blockage of DNA recognition by the innate immune system may constitute a promising therapeutic venue. (Hepatology 2015;61:348–360)

Ancillary