Graves’ disease-associated TSHR gene is demethylated and expressed in human regulatory T cells

Abstract

Epigenetic changes at specific genetic loci and the activation of transcriptional repressor FOXP3 are needed to establish and maintain the regulatory T cell (Treg) lineage. Here we studied the DNA methylation profiles in CD4+ CD25+ Tregs and CD4+ CD25- conventional T cells (Tconvs) from healthy individuals and identified a wide range of differentially methylated CpG sites (DMPs). Overall, Tregs had more hypomethylated DMPs and contained more CpG sites, which on the cell population level were less defined in their methylation status. We identified top hypomethylated CpGs in Tregs close to CENPM, IKZF2, and LYST and hypermethylated sites at the THEMIS, SCML4, and ADD3 genes. Among others, DMPs were enriched for the transcriptional repressor Kaiso binding motifs. Interestingly, in Tregs we found hypomethylation and increased expression of the TSHR gene, which is a risk gene for Graves' disease (GD). However, subsequent DNA methylation profiling in healthy individuals and GD patients revealed only 19 DMPs and no change at the TSHR locus, indicating that Tregs in GD patients share a similar methylation pattern with healthy controls. Together, we show Treg-specific hypomethylation and expression of the TSHR gene, prompting additional scenarios to explain the genetic link and role of anti-TSHR autoantibodies in GD.