Pyrrothiogatain acts as an inhibitor of GATA family proteins and inhibits Th2 cell differentiation in vitro

Shunsuke Nomura,Makoto Kuwahara,Masakatsu Yamashita,Tatsuya Sawasaki,Hirotaka Takahashi,Junpei Suzuki

doi:10.1038/s41598-019-53856-1

Abstract

The transcription factor GATA3 is a master regulator that modulates T helper 2 (Th2) cell differentiation and induces expression of Th2 cytokines, such as IL-4, IL-5, and IL-13. Th2 cytokines are involved in the protective immune response against foreign pathogens, such as parasites. However, excessive production of Th2 cytokines results in type-2 allergic inflammation. Therefore, the application of a GATA3 inhibitor provides a new therapeutic strategy to regulate Th2 cytokine production. Here, we established a novel high-throughput screening system for an inhibitor of a DNA-binding protein, such as a transcription factor, and identified pyrrothiogatain as a novel inhibitor of GATA3 DNA-binding activity. Pyrrothiogatain inhibited the DNA-binding activity of GATA3 and other members of the GATA family. Pyrrothiogatain also inhibited the interaction between GATA3 and SOX4, suggesting that it interacts with the DNA-binding region of GATA3. Furthermore, pyrrothiogatain significantly suppressed Th2 cell differentiation, without impairing Th1 cell differentiation, and inhibited the expression and production of Th2 cytokines. Our results suggest that pyrrothiogatain regulates the differentiation and function of Th2 cells via inhibition of GATA3 DNA binding activity, which demonstrates the efficiency of our drug screening system for the development of novel small compounds that inhibit the DNA-binding activity of transcription factors.

Highlights

Transcription factors are key molecules that regulate gene expression and cell fate in response to cell signalling stimuli from extracellular environments[1,2]
The pathology of allergic asthma, which is a T helper 2 (Th2) cytokine-mediated disorder, is characterized by eosinophilia, goblet cell hyperplasia, and airway smooth muscle contraction[7]. These pathological events are due to the excessive differentiation and activation of Th2 cells[8], which are a major source of Th2 cytokines, such as IL-4, IL-5, and IL-139
We selected the GATA3 transcription factor because GATA3 is known as the master regulator for Th2 cell differentiation and production of Th2 cytokines[10,11] and GATA3-binding to its DNA sequence has already been reported[11]

Summary

Introduction

Transcription factors are key molecules that regulate gene expression and cell fate in response to cell signalling stimuli from extracellular environments[1,2]. As a final step of multiple signal transductions, transcription factors regulate gene expression via nuclear translocation, dimerization, and DNA binding on the target regulatory regions in the genome. The pathology of allergic asthma, which is a Th2 cytokine-mediated disorder, is characterized by eosinophilia, goblet cell hyperplasia, and airway smooth muscle contraction[7]. These pathological events are due to the excessive differentiation and activation of Th2 cells[8], which are a major source of Th2 cytokines, such as IL-4, IL-5, and IL-139. The inhibitor screening system identified factors inhibiting the DNA-binding activity of transcription factors and facilitated the development of a novel GATA family inhibitor, pyrrothiogatain. Pyrrothiogatain suppressed in vitro Th2 cell differentiation and the secretion of Th2 cytokines without impairing Th1 cell differentiation

Methods

Results

Conclusion