Gasdermin D Hypermethylation Inhibits Pyroptosis And LPS-Induced IL-1β Release From NK92 Cells
2019
INTRODUCTION: Although natural killer (NK) are major cells used to treat cancer patients,
recent clinical trials showed that NK92 cells can be also used for the same purpose due to
their high anti-tumor activity. Here, we examined whether these cells might be inflammatory
due to the release of interleukin-1β (IL-1β), and whether the anti-inflammatory molecules
dimethyl fumarate (DMF), or monomethyl fumarate (MMF) impair this activity.
METHODS: NK92 cells were examined for the synthesis and release of IL-1β utilizing RT-PCR
and ELISA assay, respectively. The expression of hydroxy-carboxylic acid receptors (HCA)1,
HCA2 and HCA3 was detected by immunoblotting, flow cytometry, immunofluorescence and
RT-PCR assays. The activation of caspase-1 and Gasdermin D (GSDMD) was evaluated by
immunoblot assay. Pyroptosis was demonstrated by immunofluorescence imaging. Expression
of DNA methyltransferases (DNMTs) mRNA was determined by whole transcriptome and
immunoblot analyses.
RESULTS: LPS-induced the release of IL-1β from NK92 cells, whereas DMF or MMF inhibited
this induction. The effect of these drugs was due to inhibiting the conversion of procaspase-1
into active caspase-1. NK92 cells highly expressed GSDMD, a pyroptotic-mediated molecule.
However, LPS induced the distribution of GSDMD into the cell membranes, corroborated with
the presence of pyroptotic bodies, an activity that was inhibited by DMF or MMF. These
molecule also inhibited the generation of GSDMD through DNMT-mediated hypermethylation
of the promoter region of GSDMD gene. These results were supported by increased expression
of DNMTs mRNA as determined by whole transcriptome analysis.
DISCUSSION: Our results are the first to show that NK92 cells utilize GSDMD pathway to
release IL-1β. Further, DMF and MMF which were previously shown to enhance NK cell
cytotoxicity, also inhibit the inflammatory effects of these cells, making them most suitable
for treating cancer patients.
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