Increased EZH2 function in regulatory T cells promotes their capacity to suppress autoimmunity by driving effector differentiation prior to activation [RNA-seq]

The immunosuppressive function of regulatory T (Treg) cells is essential for maintaining immune homeostasis. Enhancer of zeste homolog 2 (EZH2), a histone H3 lysine 27 (H3K27) methyltransferase, plays a key role in maintaining Treg cell function upon CD28 co-stimulation, and Ezh2 deletion in Treg cells causes autoimmunity. Here we assessed whether increased EZH2 activity in Treg cells would improve Treg cell function. Using an Ezh2 gain-of-function mutation, Ezh2Y641F, we found that Treg cells expressing Ezh2Y641F displayed an increased effector Treg phenotype and were poised for improved homing to organ tissues. Expression of Ezh2Y641F in Treg cells led to more rapid remission from autoimmunity. H3K27me3 profiling and transcriptomic analysis revealed a redistribution of H3K27me3, which prompted a gene expression profile in naïve Ezh2Y641F Treg cells that recapitulated aspects of CD28-activated Ezh2WT Treg cells. Altogether, increased EZH2 activity promotes the differentiation of effector Treg cells that can better suppress autoimmunity. Overall design: RNA-seq was performed on Treg cells (sorted based on CD4+Foxp3GFP+RFP+CD62L+) from Foxp3-GFP-hCre;Ezh2Y641F/+;R26RFP (Ezh2-Y641F) or Foxp3-GFP-hCre;Ezh2+/+;R26RFP (WT) mice either collected after sorting (naive samples) or after 4 days of activation with anti-CD3 and anti-CD28 coated beads at a ratio of 1:3 (cell:bead) in the presence of 2000 IU/mL recombinant human IL-2 (activated samples).