Maintenance DNA methylation is essential for regulatory T cell development and stability of suppressive function
Kathryn A. Helmin, Luisa Morales-Nebreda, Manuel A. Torres Acosta, Kishore R. Anekalla, Shang-Yang Chen, Hiam Abdala-Valencia, Yuliya Politanska, Paul Cheresh, Mahzad Akbarpour, Elizabeth M. Steinert, Samuel E. Weinberg, Benjamin D. Singer
Kathryn A. Helmin, Luisa Morales-Nebreda, Manuel A. Torres Acosta, Kishore R. Anekalla, Shang-Yang Chen, Hiam Abdala-Valencia, Yuliya Politanska, Paul Cheresh, Mahzad Akbarpour, Elizabeth M. Steinert, Samuel E. Weinberg, Benjamin D. Singer
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Research Article Autoimmunity Immunology

Maintenance DNA methylation is essential for regulatory T cell development and stability of suppressive function

Abstract

Tregs require Foxp3 expression and induction of a specific DNA hypomethylation signature during development, after which Tregs persist as a self-renewing population that regulates immune system activation. Whether maintenance DNA methylation is required for Treg lineage development and stability and how methylation patterns are maintained during lineage self-renewal remain unclear. Here, we demonstrate that the epigenetic regulator ubiquitin-like with plant homeodomain and RING finger domains 1 (Uhrf1) is essential for maintenance of methyl-DNA marks that stabilize Treg cellular identity by repressing effector T cell transcriptional programs. Constitutive and induced deficiency of Uhrf1 within Foxp3+ cells resulted in global yet nonuniform loss of DNA methylation, derepression of inflammatory transcriptional programs, destabilization of the Treg lineage, and spontaneous inflammation. These findings support a paradigm in which maintenance DNA methylation is required in distinct regions of the Treg genome for both lineage establishment and stability of identity and suppressive function.

Authors

Kathryn A. Helmin, Luisa Morales-Nebreda, Manuel A. Torres Acosta, Kishore R. Anekalla, Shang-Yang Chen, Hiam Abdala-Valencia, Yuliya Politanska, Paul Cheresh, Mahzad Akbarpour, Elizabeth M. Steinert, Samuel E. Weinberg, Benjamin D. Singer

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Figure 5

Induced loss of Uhrf1 in Tregs results in generation of ex-Foxp3 cells.

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Induced loss of Uhrf1 in Tregs results in generation of ex-Foxp3 cells. ...
(A) Representative flow cytometry contour plots gated on splenic CD3ε+CD4+tdTomato+ cells showing the percentage of ex-Foxp3 (Foxp3-GFP–) and Treg (Foxp3-GFP+) cells after 4 weeks of tamoxifen started at either 4 or 8 weeks of age. (B) Summary data of the percentage of ex-Foxp3 and Tregs within the tdTomato+ population for the experiments shown in A. (C) Ratio of ex-Foxp3 to Tregs after 4 weeks of tamoxifen started at either 4 or 8 weeks of age. For mice started on tamoxifen at 4 weeks of age, n = 5 (iUhrf1+/+) and n = 18 (iUhrf1fl/fl); for mice started on tamoxifen at 8 weeks of age, n = 4 (iUhrf1+/+) and n = 8 (iUhrf1fl/fl). (D) Total cell numbers of ex-Foxp3 and Tregs. For mice started on tamoxifen at 4 weeks of age, n = 5 (iUhrf1+/+) and n = 7 (iUhrf1fl/fl); for mice started on tamoxifen at 8 weeks of age, n = 4 (iUhrf1+/+) and n = 7 (iUhrf1fl/fl). *q < 0.05; **p or q < 0.01; †p or q < 0.001; NS, not significant by Mann-Whitney U test (C) or the 2-stage linear step-up procedure of Benjamini, Krieger, and Yekutieli with Q = 5% (B and D); exact p and q values are in Supplemental Data.