Hepatic DNA deposition drives drug‐induced liver injury and ... : Hepatology (original) (raw)

Liver Injury/Regeneration

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

Marques, Pedro Elias1,†; Oliveira, André Gustavo1,2,†; Pereira, Rafaela Vaz1; David, Bruna Araújo1; Gomides, Lindisley Ferreira1; Saraiva, Adriana Machado1; Pires, Daniele Araújo1; Novaes, Júlia Tosta1; Patricio, Daniel O.3; Cisalpino, Daniel4; Menezes‐Garcia, Zélia3; Leevy, W. Matthew5; Chapman, Sarah Ellen5; Mahecha, GermánArturo6; Marques, Rafael Elias7; Guabiraba, Rodrigo8; Martins, Vicente Paulo9; Souza, Danielle Gloria4; Mansur, Daniel Santos3; Teixeira, Mauro Martins8; Leite, M. Fatima2; Menezes, Gustavo Batista*,1

1Laboratório de Imunobiofotônica, Departamento de Morfologia, ICB, Universidade Federal de Minas GeraisMinas GeraisBrazil

2Departamento de Fisiologia e BiofísicaICB, Universidade Federal de Minas GeraisMinas GeraisBrazil

3Departamento de Microbiologia, Imunologia e ParasitologiaCCB, Universidade Federal de Santa CatarinaSanta CatarinaBrazil

4Departamento de MicrobiologiaICB, Universidade Federal de Minas GeraisMinas GeraisBrazil

5Biological Imaging CoreUniversity of Notre Dame, Department of Biological SciencesSouth BendINUSA

6Departamento de MorfologiaICB, Universidade Federal de Minas GeraisMinas GeraisBrazil

7Laboratório de Imunofarmacologia, Departamento de Bioquímica e ImunologiaICB, Universidade Federal de Minas GeraisMinas GeraisBrazil

8INRA, Pôle Santé Animale de Tours PSAT, UMR 1282, Infectiologie et Santé PubliqueISP 213Nouzilly37380France

9Departamento de Biologia Celular (CEL)Instituto de Ciências Biológicas (IB), Universidade de Brasília (UnB)BrasíliaDFBrazil

*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: [email protected]

†These authors contributed equally to this work.

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)