CD4+FOXP3+lymphocytes are expanded within the intestinal inflammatory lesion of Crohn?s disease (CD); however, ongoing inflammation belies presumed anti-inflammatory function of this cell. FOXP3 is required for differentiation and function of T regulatory (TREG) cells. FOXP3-mediated gene repression is lost in intestinal T cells of CD patients. Indeed, ?FOXP3+ Crohn?s cell (FOXP3+CD)? bears a transcriptional signal more closely related to the pro-inflammatory TH17 cell. Derivation, function, and therapeutic implications of the FOXP3+CD cell remain poorly understood. Our long-term goal is to dissect epigenetic mechanisms regulating TREG cellular differentiation and function in the setting of GI inflammation. Consequently, the objective is to identify mechanisms responsible for activation of the TH17 phenotypic transcriptional network within intestinal FOXP3+CD cells and test therapeutic opportunities to restore their regulatory function in vivo. Our central hypothesis is that a shared chromatin configuration between TREG and TH17 cells allows for rapid activation of a TH17-like pro-inflammatory gene program in intestinal FOXP3+TREG cells. A set of TH17-relevant genes are accessible, yet not active in human TREG cells. A defining feature of the TH17 program was significant enrichment of CCCTC binding factor (CTCF) motifs. CTCF, a well-known topologically-associated domain (TAD) insulator protein, also mediates intra-TAD chromosomal looping and enhancer-promoter interaction regulating gene transcription. Bioinformatic analysis of this TH17 program indicates an inhibitory role for Polycomb Repressor Complex 1 (PRC1). Deletion of PRC1 in murine FOXP3+ cells led to secretion of prototypic TH17-like cytokines in FOXP3+ cells, and spontaneous colitis. The rationale is that with mechanistic insight into the biology of intestinal TREG cell in inflammation, one can apply targeted FOXP3+ CD cell-directed therapy including engineering of human TREG cells for adoptive cell therapy trials. To test the central hypothesis and obtain the overall objective we will address the following three questions:
Aim1 : Why are CTCF motifs associated with the TH17 program? Aim2: Why are TH17-associated genes accessible yet not active in TREG cells? Aim3: Why do FOXP3+CD cells express prototypic TH17 cytokines? Upon conclusion, we will understand the epigenetic pathways and 3D chromatin architecture spawning the FOXP3+CD cell and mechanisms shaping their development and function. This contribution is significant as the FOXP3+CD cell is expanded in the Crohn?s lesion, phenocopies TH17 cell which is implicated in Crohn?s disease (and other inflammatory diseases), and represents an important epigenetic drug target for systemic or re-engineered cellular therapy. There is general acknowledgement of risk of FOXP3+ TREG cells converting to pathogenic FOXP3+TH17-like cells upon adoptive transfer in Crohn?s trials; yet, this project is the first to articulate both signature and precise epigenetic events that regulate activation of the TH17 program in TREG cells.

Public Health Relevance

The proposed research is relevant to public health because it focuses on the development and maintenance of human FOXP3+TREG and TH17 cells, work which will open novel experimental therapeutics including cellular therapy in human Inflammatory Bowel Disease (IBD) and other TH17-related inflammatory diseases (i.e. rheumatologic, neurologic, and dermatologic conditions). Ongoing TREG cell therapy trials and emerging cancer trials of epigenetic regulators will benefit from insight into potential activation of the TH17 phenotype in FOXP3+ cells exposed to the inflammatory milieu. We believe that execution of the experimental plan will realize the new horizon of epigenetic therapies in human immune mediated diseases.

National Institute of Health (NIH)
National Institute of Allergy and Infectious Diseases (NIAID)
Research Project (R01)
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Special Emphasis Panel (ZRG1)
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Rothermel, Annette L
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Mayo Clinic, Rochester
United States
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Kosinsky, Robyn Laura; Chua, Robert Lorenz; Qui, Martin et al. (2018) Loss of RNF40 decreases NF-?B activity in colorectal cancer cells and reduces colitis burden in mice. J Crohns Colitis :
Dietz, Allan B; Dozois, Eric J; Fletcher, Joel G et al. (2017) Autologous Mesenchymal Stem Cells, Applied in a Bioabsorbable Matrix, for Treatment of Perianal Fistulas in Patients With Crohn's Disease. Gastroenterology 153:59-62.e2
Sarmento, Olga F; Svingen, Phyllis A; Xiong, Yuning et al. (2017) The Role of the Histone Methyltransferase Enhancer of Zeste Homolog 2 (EZH2) in the Pathobiological Mechanisms Underlying Inflammatory Bowel Disease (IBD). J Biol Chem 292:706-722
Velez, Gabriel; Lin, Marisa; Christensen, Trace et al. (2016) Evidence supporting a critical contribution of intrinsically disordered regions to the biochemical behavior of full-length human HP1?. J Mol Model 22:12
Mathison, Angela; Escande, Carlos; Calvo, Ezequiel et al. (2015) Phenotypic Characterization of Mice Carrying Homozygous Deletion of KLF11, a Gene in Which Mutations Cause Human Neonatal and MODY VII Diabetes. Endocrinology 156:3581-95
Sarmento, Olga F; Svingen, Phyllis A; Xiong, Yuning et al. (2015) A novel role for KLF14 in T regulatory cell differentiation. Cell Mol Gastroenterol Hepatol 1:188-202.e4
Papadakis, Konstantinos A; Krempski, James; Reiter, Jesse et al. (2015) Krüppel-like factor KLF10 regulates transforming growth factor receptor II expression and TGF-? signaling in CD8+ T lymphocytes. Am J Physiol Cell Physiol 308:C362-71
Dave, Maneesh; Hayashi, Yujiro; Gajdos, Gabriella B et al. (2015) Stem cells for murine interstitial cells of cajal suppress cellular immunity and colitis via prostaglandin E2 secretion. Gastroenterology 148:978-90
Papadakis, Konstantinos A; Krempski, James; Svingen, Phyllis et al. (2015) Krüppel-like factor KLF10 deficiency predisposes to colitis through colonic macrophage dysregulation. Am J Physiol Gastrointest Liver Physiol 309:G900-9
Dave, Maneesh; Mehta, Kathan; Luther, Jay et al. (2015) Mesenchymal Stem Cell Therapy for Inflammatory Bowel Disease: A Systematic Review and Meta-analysis. Inflamm Bowel Dis 21:2696-707

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