Author:
Araujo Alan,Antunes Maísa,Mattos Matheus,Diniz Ariane,Alvarenga Débora,Nakagaki Brenda,Carvalho Érika,Lacerda Viviane,Carvalho-Gontijo Raquel,Goulart Jorge,Mafra Kassiana,Freitas-Lopes Maria,Oliveira Hortência,Dutra Camila,David Bruna,Mendes Silva Aristóbolo,Quesniaux Valerie,Ryffel Bernhard,Oliveira Sergio,Barber Glen,Mansur Daniel,Cunha Thiago,Rezende Rafael,Oliveira André,Menezes Gustavo
Abstract
Hepatocytes may rupture after a drug overdose, and their intracellular contents act as damage-associated molecular patterns (DAMPs) that lead to additional leukocyte infiltration, amplifying the original injury. Necrosis-derived DNA can be recognized as a DAMP, activating liver non-parenchymal cells (NPCs). We hypothesized that NPCs react to DNA by releasing interferon (IFN)-1, which amplifies acetaminophen (APAP)-triggered liver necrosis. We orally overdosed different knockout mouse strains to investigate the pathways involved in DNA-mediated amplification of APAP-induced necrosis. Mice were imaged under intravital confocal microscopy to estimate injury progression, and hepatocytes and liver NPCs were differentially isolated for gene expression assays. Flow cytometry (FACS) using a fluorescent reporter mouse estimated the interferon-beta production by liver leukocytes under different injury conditions. We also treated mice with DNase to investigate the role of necrosis DNA signaling in IFN-1 production. Hepatocytes released a large amount of DNA after APAP overdose, which was not primarily sensed by these cells. However, liver NPCs promptly sensed such environmental disturbances and activated several DNA sensing pathways. Liver NPCs synthesized and released IFN-1, which was associated with concomitant hepatocyte necrosis. Ablation of IFN-1 recognition in interferon α/β receptor (IFNAR−/−) mice delayed APAP-mediated liver necrosis and dampened IFN-1 sensing pathways. We demonstrated a novel loop involving DNA recognition by hepatic NPCs and additional IFN-1 mediated hepatocyte death.
Funder
Conselho Nacional de Desenvolvimento Científico e Tecnológico
Fundação de Amparo à Pesquisa do Estado de Minas Gerais
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior