# Epigenetically Controlled ZEB2 Expression Promotes the Cytotoxic Potential of CMV‐Specific CD8+ T Cells

## TL;DR

This paper shows that ZEB2 gene expression, controlled by DNA methylation changes, is important for the cytotoxic function of virus-specific CD8+ T cells.

## Contribution

The study reveals that ZEB2 expression is epigenetically regulated and essential for the cytotoxic function of CMV-specific CD8+ T cells.

## Key findings

- ZEB2 differentially methylated regions (DMRs) undergo demethylation during T cell differentiation.
- ZEB2 knockout reduces cytotoxic capacity in CMV-specific CD8+ T cells.
- ZEB2 expression correlates with T cell effector and memory differentiation.

## Abstract

Zinc finger E‐box binding protein 2 (ZEB2) is a key factor in the differentiation of naïve CD8+ T cells into effector and memory T cells. However, the precise regulatory role of ZEB2 in cytotoxic CD8+ T cells remains unknown. Our recent DNA methylation analysis of cytomegalovirus (CMV)‐specific human CD8+ T cells revealed two differentially methylated regions (DMRs) within the ZEB2 locus. In the present study, we show that these ZEB2 DMRs undergo pronounced demethylation during T cell differentiation. In particular, terminally differentiated CD8+ T cells and cytotoxic CD4+ T cells show an almost complete demethylation. Demethylation of the ZEB2 DMRs correlates strongly with ZEB2 expression in all T cell subsets. Furthermore, DNA methylation patterns remain stable during long‐term in vitro culture. ZEB2 knockout in CD8+ effector T cells results in altered gene expression profiles, affecting genes related to cell–cell adhesion and impairing the cytotoxic capacity in CMV‐specific killing assays. Our data show that ZEB2 expression contributes to the differentiation of naïve CD8+ T cells into effector and memory T cells and regulates the functional properties of virus‐specific cytotoxic CD8+ T cells.

Differentially methylated regions (DMRs) within the ZEB2 locus progressively demethylate during CD4+ and CD8+ T cell differentiation into cytotoxic CD4+ and CD8+ TEMRA subsets, respectively. This epigenetic remodeling tightly correlates with elevated ZEB2 expression, and ZEB2 knockout (KO) impairs the effector and cytotoxic function of CD4+ and CD8+ T cells.