DNA demethylation increases NETosis

Hiroyuki Yasuda; Yutaka Takishita; Akihiro Morita; Tomonari Tsutsumi; Masahiko Tsuchiya; Eisuke F Sato

doi:10.1016/j.abb.2020.108465

DNA demethylation increases NETosis

Arch Biochem Biophys. 2020 Aug 15:689:108465. doi: 10.1016/j.abb.2020.108465. Epub 2020 Jun 17.

Authors

Hiroyuki Yasuda¹, Yutaka Takishita¹, Akihiro Morita¹, Tomonari Tsutsumi¹, Masahiko Tsuchiya², Eisuke F Sato³

Affiliations

¹ Department of Biochemistry, Faculty of Pharmaceutical Sciences, Suzuka University of Medical Science, 3500-3, Minamitamagaki, Suzuka-city, Mie, 513-8670, Japan.
² Department of Anesthesiology, Osaka City University Medical School, 1-5-7, Asahi-machi, Abeno, Osaka, 545-8586, Japan.
³ Department of Biochemistry, Faculty of Pharmaceutical Sciences, Suzuka University of Medical Science, 3500-3, Minamitamagaki, Suzuka-city, Mie, 513-8670, Japan. Electronic address: efsato@suzuka-u.ac.jp.

PMID: 32561201
DOI: 10.1016/j.abb.2020.108465

Abstract

Neutrophil extracellular traps (NETs) occur during the development of autoimmune diseases, cancer and diabetes. A novel form of cell death that is induced by NETs is called NETosis. Although these diseases are known to have an epigenetic component, epigenetic regulation of NETosis has not previously been explored. In the present study, we investigated the effects of epigenetic change, especially DNA demethylation, on NETosis in neutrophil-like cells differentiated from HL-60 cells, which were incubated for 72 h in the presence of 1.25% DMSO. DMSO-differentiated neutrophil-like cells tended to have increased methylation of genomic DNA. NETosis in the neutrophil-like cells was induced by the treatment with A23187, calcium ionophore, and increased by the addition of the DNMT inhibitor 5-azacytidine (Aza) during differentiation. Interestingly, Aza-stimulated neutrophil-like cell induced NETosis without treatment with A23187. Although reactive oxygen species (ROS), especially superoxide and hypochlorous acid, are important in NETosis induction, treatment with Aza decreased production of ROS, while mitochondria ROS scavenger tended to decrease Aza-induced NETosis. Moreover, the genomic DNA in Aza-stimulated neutrophil-like cell was demethylated, and the expression of peptidylarginine deiminase4 (PAD4) and citrullinated histone H3 (R2+R8+R17) was increased, but myeloperoxidase expression was unaffected. Additionally, PAD4 inhibition tended to decrease Aza-induced NETosis. The DNA demethylation induced by the DNMT inhibitor in neutrophil-like cells enhanced spontaneous NETosis through increasing PAD4 expression and histone citrullination. This study establishes a relationship between NETosis and epigenetics for the first time, and indicates that various diseases implicated to have an epigenetic component might be exacerbated by excessive NETosis also under epigenetic control.

Keywords: DNA methylation; Histone citrullination; NETosis; Neutrophil extracellular trap; Peptidylarginine deiminase 4.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Death*
Cell Differentiation
DNA / genetics
DNA Demethylation*
Epigenesis, Genetic
Extracellular Traps / genetics*
HL-60 Cells
Humans
Neutrophils / cytology*
Neutrophils / metabolism

Substances

DNA