Disruption of FOXP3–EZH2 Interaction Represents a Pathobiological Mechanism in Intestinal Inflammation

Adebowale O. Bamidele, Phyllis A. Svingen, Mary R. Sagstetter, Olga F. Sarmento, Michelle Gonzalez, Manuel B. Braga Neto, Subra Kugathasan, Gwen Lomberk, Raul A. Urrutia, William Alvis Faubion

Research output: Contribution to journalArticle

Abstract

Background & Aims: Forkhead box protein 3 (FOXP3)+ regulatory T cell (Treg) dysfunction is associated with autoimmune diseases; however, the mechanisms responsible for inflammatory bowel disease pathophysiology are poorly understood. Here, we tested the hypothesis that a physical interaction between transcription factor FOXP3 and the epigenetic enzyme enhancer of zeste homolog 2 (EZH2) is essential for gene co-repressive function. Methods: Human FOXP3 mutations clinically relevant to intestinal inflammation were generated by site-directed mutagenesis. T lymphocytes were isolated from mice, human blood, and lamina propria of Crohn's disease (CD) patients and non-CD controls. We performed proximity ligation or a co-immunoprecipitation assay in FOXP3-mutant+, interleukin 6 (IL6)-treated or CD-CD4+ T cells to assess FOXP3–EZH2 protein interaction. We studied IL2 promoter activity and chromatin state of the interferon γ locus via luciferase reporter and chromatin-immunoprecipitation assays, respectively, in cells expressing FOXP3 mutants. Results: EZH2 binding was abrogated by inflammatory bowel disease–associated FOXP3 cysteine 232 (C232) mutation. The C232 mutant showed impaired repression of IL2 and diminished EZH2-mediated trimethylation of histone 3 at lysine 27 on interferon γ, indicative of compromised Treg physiologic function. Generalizing this mechanism, IL6 impaired FOXP3–EZH2 interaction. IL6-induced effects were reversed by Janus kinase 1/2 inhibition. In lamina propria–derived CD4+T cells from CD patients, we observed decreased FOXP3–EZH2 interaction. Conclusions: FOXP3–C232 mutation disrupts EZH2 recruitment and gene co-repressive function. The proinflammatory cytokine IL6 abrogates FOXP3–EZH2 interaction. Studies in lesion-derived CD4+ T cells have shown that reduced FOXP3–EZH2 interaction is a molecular feature of CD patients. Destabilized FOXP3–EZH2 protein interaction via diverse mechanisms and consequent Treg abnormality may drive gastrointestinal inflammation.

LanguageEnglish (US)
Pages55-71
Number of pages17
JournalCMGH
Volume7
Issue number1
DOIs
StatePublished - Jan 1 2019

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Forkhead Transcription Factors
Inflammation
Crohn Disease
Interleukin-6
Cysteine
T-Lymphocytes
Inflammatory Bowel Diseases
Enhancer of Zeste Homolog 2 Protein
Mutation
Interferon-gamma
Interleukin-2
Janus Kinase 1
Mucous Membrane
Janus Kinase 2
Chromatin Immunoprecipitation
Essential Genes
Regulatory T-Lymphocytes
Site-Directed Mutagenesis
Luciferases
Immunoprecipitation

Keywords

  • Crohn's Disease
  • Epigenetics
  • Proinflammatory Cytokine
  • Regulatory T Cells

ASJC Scopus subject areas

  • Hepatology
  • Gastroenterology

Cite this

Bamidele, A. O., Svingen, P. A., Sagstetter, M. R., Sarmento, O. F., Gonzalez, M., Braga Neto, M. B., ... Faubion, W. A. (2019). Disruption of FOXP3–EZH2 Interaction Represents a Pathobiological Mechanism in Intestinal Inflammation. CMGH, 7(1), 55-71. https://doi.org/10.1016/j.jcmgh.2018.08.009

Disruption of FOXP3–EZH2 Interaction Represents a Pathobiological Mechanism in Intestinal Inflammation. / Bamidele, Adebowale O.; Svingen, Phyllis A.; Sagstetter, Mary R.; Sarmento, Olga F.; Gonzalez, Michelle; Braga Neto, Manuel B.; Kugathasan, Subra; Lomberk, Gwen; Urrutia, Raul A.; Faubion, William Alvis.

In: CMGH, Vol. 7, No. 1, 01.01.2019, p. 55-71.

Research output: Contribution to journalArticle

Bamidele, AO, Svingen, PA, Sagstetter, MR, Sarmento, OF, Gonzalez, M, Braga Neto, MB, Kugathasan, S, Lomberk, G, Urrutia, RA & Faubion, WA 2019, 'Disruption of FOXP3–EZH2 Interaction Represents a Pathobiological Mechanism in Intestinal Inflammation', CMGH, vol. 7, no. 1, pp. 55-71. https://doi.org/10.1016/j.jcmgh.2018.08.009
Bamidele AO, Svingen PA, Sagstetter MR, Sarmento OF, Gonzalez M, Braga Neto MB et al. Disruption of FOXP3–EZH2 Interaction Represents a Pathobiological Mechanism in Intestinal Inflammation. CMGH. 2019 Jan 1;7(1):55-71. https://doi.org/10.1016/j.jcmgh.2018.08.009
Bamidele, Adebowale O. ; Svingen, Phyllis A. ; Sagstetter, Mary R. ; Sarmento, Olga F. ; Gonzalez, Michelle ; Braga Neto, Manuel B. ; Kugathasan, Subra ; Lomberk, Gwen ; Urrutia, Raul A. ; Faubion, William Alvis. / Disruption of FOXP3–EZH2 Interaction Represents a Pathobiological Mechanism in Intestinal Inflammation. In: CMGH. 2019 ; Vol. 7, No. 1. pp. 55-71.
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AU - Sarmento, Olga F.

AU - Gonzalez, Michelle

AU - Braga Neto, Manuel B.

AU - Kugathasan, Subra

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AU - Urrutia, Raul A.

AU - Faubion, William Alvis

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