Project Details
Projekt
Functional role of DNA Methylation in human regulatory T cells.
Applicant
Professorin Dr. Julia Polansky-Biskup
Subject Area
Cell Biology
Immunology
Immunology
Term
from 2017 to 2021
Project identifier
Deutsche Forschungsgemeinschaft (DFG) - Project number 388741410
The generation and maintenance of CD4+ regulatory T cells (Tregs) is guided by Treg-specific differentially methylated regions (Treg-DMRs) which act as epigenetic regulator elements and control the Treg-specific transcriptional profile. The demethylated state of a Treg-DMR usually correlates with expression of the target gene. However, proof for a causal relationship between DMR de-methylation and target gene expression in the endogenous chromatin context of a living cell is so far lacking, mainly due to technical limitations. This also holds true for the Treg-DMR 'TSDR', which is regulating expression of the Treg-specific transcription factor FOXP3.In the present project, we want to modify the methylation state of the TSDR in primary human T cells using various CRISPR/Cas9-mediated 'epi-/genetic editing' methods and analyze the effects on TSDR activity and on the Treg-phenotype. With this, we want to clarify whether DNA demethylation is causal for TSDR activity and whether certain key CpG motifs are of special importance. In addition, the influence of the endogenous chromatin structure on the maintenance of the TSDR methylation state should be clarified. By analyzing additional Treg-DMRs for causality and chromatin influence, we want to clarify whether both features are of general importance or dependent on the target gene. We hope to be able to proof for the first time a causal relationship between the methylation state and the transcriptional activity of Treg-DMRs and its implications for the Treg-phenotype. In this effort, we might additionally identify cell type-determining DMRs or key CpGs which could gain clinical relevance in the future, mainly in epigenetic editing approaches for the induction of therapeutic Treg populations.
DFG Programme
Research Grants
