Tregs epigenetically reprogrammed from autoreactive effector T cells mitigate established autoimmunity
Tyler R. Colson, James J. Cameron, Hayley I. Muendlein, Mei-An Nolan, Jamie L. Leiriao, James H. Kim, Alexander N. Poltorak, Xudong Li
Tyler R. Colson, James J. Cameron, Hayley I. Muendlein, Mei-An Nolan, Jamie L. Leiriao, James H. Kim, Alexander N. Poltorak, Xudong Li
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Research Article Immunology Inflammation

Tregs epigenetically reprogrammed from autoreactive effector T cells mitigate established autoimmunity

Abstract

Reprogramming autoreactive CD4+ effector T (Teff) cells into immunosuppressive Tregs represents a promising strategy for treating established autoimmune diseases. However, the stability and function of such reprogrammed Tregs under inflammatory conditions remain unclear. Here, we show that demethylation of core Treg identity genes in Teff cells yields lineage-stable effector T cell reprogrammed Tregs (ER-Tregs). A single adoptive transfer of ER-Tregs not only prevents autoimmune neuroinflammation in mice when given before disease onset but also arrests its progression when administered after onset. Compared with Foxp3-overexpressing Teff cells, induced Tregs from naive precursors, and endogenous Tregs, ER-Tregs provide superior protection against autoimmune neuroinflammation. This enhanced efficacy stems from their inherited autoantigen specificity and selectively preserved effector cell transcriptional programs, which together bolster their fitness in inflammatory environments and enhance their suppressive capacity. Our results establish epigenetic reprogramming of autoreactive Teff cells as an effective approach to generate potent, stable Tregs for the treatment of refractory autoimmune conditions.

Authors

Tyler R. Colson, James J. Cameron, Hayley I. Muendlein, Mei-An Nolan, Jamie L. Leiriao, James H. Kim, Alexander N. Poltorak, Xudong Li

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Figure 7

Elevated expression of specific Teff genes contributes to ER-Treg fitness and suppressive function in EAE.

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Elevated expression of specific Teff genes contributes to ER-Treg fitnes...
(A–C) EAE was induced in Rag1–/– mice 1 day after transfer of MOG/CFA-primed CD45.1+ CD4+ Tconv cells with or without cotransfer of CD45.2+ ER-Tregs or CD45.2+ nTregs isolated from MOG/CFA-primed mice and cultured in the presence of IL-2 for 3 days. Flow cytometry analyses were conducted at 22 dpi. n = 6 per group. (A) EAE disease curve. (B) Flow cytometry of transferred Treg frequencies. (C) Flow cytometry of RORγt expression in transferred Tregs. (D) Flow cytometry of in vivo competitive fitness between fluorescent reporter transduced 2D2 ER-Tregs and nTregs, cotransferred at a 1:1 ratio 1 day prior to CFA/MOG immunization and analyzed 5 dpi. (E–I) EAE was induced in Rag1–/– mice 1 day after transfer of MOG/CFA-primed CD45.1+ CD4+ Tconv cells with or without cotransfer of CD45.2+Foxp3Thy1.1R26Cas9 ER-Tregs transduced with sgRNA-RV targeting Stat3 (sgStat3), Maf (sgMaf), or a nontargeting sgRNA-RV (sgNT). Flow cytometry analyses were conducted at 22 dpi. n = 6–7 per group. (E) EAE disease curve. (F) Flow cytometry of the frequencies (left) and numbers (right) of CD4+ T cells in the spinal cord. (G) Numbers of GM-CSF+ CD4+ T cells in the spinal cord. (H) Flow cytometry of c-Maf and RORγt expression in draining LN ER-Tregs.. (I) Flow cytometry of the frequencies and numbers of transferred ER-Tregs in CD4+ T cells in the spinal cord (upper) and draining LNs (lower). (J) Flow cytometry of IFN-γ and GM-CSF expression in CTVloCD4+ Teff cells cocultured for 3 days with APCs and MOG in the presence or absence of sgMaf-RV or sgNT-RV transduced Foxp3Thy1.1R26Cas9 ER-Tregs. Data are shown as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, 1-way ANOVA and Holm-Šídák test in A, E, and F–J and unpaired 2-tailed t test in B–D.