Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
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DNA Demethylation of the Foxp3 Enhancer Is Maintained through Modulation of Ten-Eleven-Translocation and DNA Methyltransferases

  • Nair, Varun Sasidharan (Department of Pathology, Hallym University College of Medicine) ;
  • Song, Mi Hye (Department of Pathology, Hallym University College of Medicine) ;
  • Ko, Myunggon (School of Life Sciences, Ulsan National Institute of Science and Technology) ;
  • Oh, Kwon Ik (Department of Pathology, Hallym University College of Medicine)
  • Received : 2016.11.15
  • Accepted : 2016.11.24
  • Published : 2016.12.31
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Abstract

Stable expression of Foxp3 is ensured by demethylation of CpG motifs in the Foxp3 intronic element, the conserved non-coding sequence 2 (CNS2), which persists throughout the lifespan of regulatory T cells (Tregs). However, little is known about the mechanisms on how CNS2 demethylation is sustained. In this study, we found that Ten-Eleven-Translocation (Tet) DNA dioxygenase protects the CpG motifs of CNS2 from re-methylation by DNA methyltransferases (Dnmts) and prevents Tregs from losing Foxp3 expression under inflammatory conditions. Upon stimulation of Tregs by interleukin-6 (IL6), Dnmt1 was recruited to CNS2 and induced methylation, which was inhibited by Tet2 recruited by IL2. Tet2 prevented CNS2 re-methylation by not only the occupancy of the CNS2 locus but also by its enzymatic activity. These results show that the CNS2 methylation status is dynamically regulated by a balance between Tets and Dnmts which influences the expression of Foxp3 in Tregs.

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References

  1. Burzyn, D., Kuswanto, W., Kolodin, D., Shadrach, J.L., Cerletti, M., Jang, Y., Sefik, E., Tan, T.G., Wagers, A.J., Benoist, C., et al. (2013). A special population of regulatory T cells potentiates muscle repair. Cell 155, 1282-1295. https://doi.org/10.1016/j.cell.2013.10.054
  2. Campbell, D.J., and Koch, M.A. (2011). Phenotypical and functional specialization of FOXP3+ regulatory T cells. Nat. Rev. Immunol. 11, 119-130. https://doi.org/10.1038/nri2916
  3. Chaudhry, A., Rudra, D., Treuting, P., Samstein, R.M., Liang, Y., Kas, A., and Rudensky, A.Y. (2009). CD4+ regulatory T cells control TH17 responses in a Stat3-dependent manner. Science 326, 986-991. https://doi.org/10.1126/science.1172702
  4. Chung, Y., Tanaka, S., Chu, F., Nurieva, R.I., Martinez, G.J., Rawal, S., Wang, Y.H., Lim, H., Reynolds, J.M., Zhou, X.H., et al. (2011). Follicular regulatory T cells expressing Foxp3 and Bcl-6 suppress germinal center reactions. Nat. Med. 17, 983-988. https://doi.org/10.1038/nm.2426
  5. Cipolletta, D., Feuerer, M., Li, A., Kamei, N., Lee, J., Shoelson, S.E., Benoist, C., and Mathis, D. (2012). PPAR-gamma is a major driver of the accumulation and phenotype of adipose tissue Treg cells. Nature 486, 549-553. https://doi.org/10.1038/nature11132
  6. Dickson, K.M., Gustafson, C.B., Young, J.I., Zuchner, S., and Wang, G. (2013). Ascorbate-induced generation of 5-hydroxymethylcytosine is unaffected by varying levels of iron and 2-oxoglutarate. Biochem. Biophys. Res. Commun. 439, 522-527. https://doi.org/10.1016/j.bbrc.2013.09.010
  7. Feng, Y., Arvey, A., Chinen, T., van der Veeken, J., Gasteiger, G., and Rudensky, A.Y. (2014). Control of the inheritance of regulatory T cell identity by a cis element in the Foxp3 locus. Cell 158, 749-763. https://doi.org/10.1016/j.cell.2014.07.031
  8. Floess, S., Freyer, J., Siewert, C., Baron, U., Olek, S., Polansky, J., Schlawe, K., Chang, H.D., Bopp, T., Schmitt, E., et al. (2007). Epigenetic control of the foxp3 locus in regulatory T cells. PLoS Biol. 5, e38. https://doi.org/10.1371/journal.pbio.0050038
  9. Fontenot, J.D., Gavin, M.A., and Rudensky, A.Y. (2003). Foxp3 programs the development and function of CD4+CD25+ regulatory T cells. Nat. Immunol. 4, 330-336. https://doi.org/10.1038/ni904
  10. Han, J.A., An, J., and Ko, M. (2015). Functions of TET proteins in hematopoietic transformation. Mol. Cells 38, 925-935.
  11. Hansmann, L., Schmidl, C., Boeld, T.J., Andreesen, R., Hoffmann, P., Rehli, M., and Edinger, M. (2010). Isolation of intact genomic DNA from FOXP3-sorted human regulatory T cells for epigenetic analyses. Eur. J. Immunol. 40, 1510-1512. https://doi.org/10.1002/eji.200940154
  12. Hori, S., Nomura, T., and Sakaguchi, S. (2003). Control of regulatory T cell development by the transcription factor Foxp3. Science 299, 1057-1061. https://doi.org/10.1126/science.1079490
  13. Huehn, J., and Beyer, M. (2015). Epigenetic and transcriptional control of Foxp3+ regulatory T cells. Semin. Immunol. 27, 10-18. https://doi.org/10.1016/j.smim.2015.02.002
  14. Huehn, J., Polansky, J.K., and Hamann, A. (2009). Epigenetic control of FOXP3 expression: the key to a stable regulatory T-cell lineage? Nat. Rev. Immunol. 9, 83-89. https://doi.org/10.1038/nri2474
  15. Jeltsch, A., and Jurkowska, R.Z. (2014). New concepts in DNA methylation. Trends Biochem. Sci. 39, 310-318. https://doi.org/10.1016/j.tibs.2014.05.002
  16. Josefowicz, S.Z., Wilson, C.B., and Rudensky, A.Y. (2009). Cutting edge: TCR stimulation is sufficient for induction of Foxp3 expression in the absence of DNA methyltransferase 1. J. Immunol. 182, 6648-6652. https://doi.org/10.4049/jimmunol.0803320
  17. Kim, H.P., and Leonard, W.J. (2007). CREB/ATF-dependent T cell receptor-induced FoxP3 gene expression: a role for DNA methylation. J. Exp. Med. 204, 1543-1551. https://doi.org/10.1084/jem.20070109
  18. Kim, K.Y., Lee, G., Yoon, M., Cho, E.H., Park, C.S., and Kim, M.G. (2015). Expression analyses revealed thymic stromal cotransporter/ Slc46A2 is in stem cell populations and is a putative tumor suppressor. Mol. Cells 38, 548-561. https://doi.org/10.14348/molcells.2015.0044
  19. Ko, M., Huang, Y., Jankowska, A.M., Pape, U.J., Tahiliani, M., Bandukwala, H.S., An, J., Lamperti, E.D., Koh, K.P., Ganetzky, R., et al. (2010). Impaired hydroxylation of 5-methylcytosine in myeloid cancers with mutant TET2. Nature 468, 839-843. https://doi.org/10.1038/nature09586
  20. Koch, M.A., Tucker-Heard, G., Perdue, N.R., Killebrew, J.R., Urdahl, K.B., and Campbell, D.J. (2009). The transcription factor T-bet controls regulatory T cell homeostasis and function during type 1 inflammation. Nat. Immunol. 10, 595-602. https://doi.org/10.1038/ni.1731
  21. Li, X., Liang, Y., LeBlanc, M., Benner, C., and Zheng, Y. (2014). Function of a Foxp3 cis-element in protecting regulatory T cell identity. Cell 158, 734-748. https://doi.org/10.1016/j.cell.2014.07.030
  22. Linterman, M.A., Pierson, W., Lee, S.K., Kallies, A., Kawamoto, S., Rayner, T.F., Srivastava, M., Divekar, D.P., Beaton, L., Hogan, J.J., et al. (2011). Foxp3+ follicular regulatory T cells control the germinal center response. Nat. Med. 17, 975-982. https://doi.org/10.1038/nm.2425
  23. Malek, T.R. (2008). The biology of interleukin-2. Annu. Rev. Immunol. 26, 453-479. https://doi.org/10.1146/annurev.immunol.26.021607.090357
  24. Mantei, A., Rutz, S., Janke, M., Kirchhoff, D., Jung, U., Patzel, V., Vogel, U., Rudel, T., Andreou, I., Weber, M., et al. (2008). siRNA stabilization prolongs gene knockdown in primary T lymphocytes. Eur. J. Immunol. 38, 2616-2625. https://doi.org/10.1002/eji.200738075
  25. Miyao, T., Floess, S., Setoguchi, R., Luche, H., Fehling, H.J., Waldmann, H., Huehn, J., and Hori, S. (2012). Plasticity of Foxp3(+) T cells reflects promiscuous Foxp3 expression in conventional T cells but not reprogramming of regulatory T cells. Immunity 36, 262-275. https://doi.org/10.1016/j.immuni.2011.12.012
  26. Moran-Crusio, K., Reavie, L., Shih, A., Abdel-Wahab, O., Ndiaye-Lobry, D., Lobry, C., Figueroa, M.E., Vasanthakumar, A., Patel, J., Zhao, X., et al. (2011). Tet2 loss leads to increased hematopoietic stem cell self-renewal and myeloid transformation. Cancer Cell 20, 11-24. https://doi.org/10.1016/j.ccr.2011.06.001
  27. Ohkura, N., Hamaguchi, M., Morikawa, H., Sugimura, K., Tanaka, A., Ito, Y., Osaki, M., Tanaka, Y., Yamashita, R., Nakano, N., et al. (2012). T cell receptor stimulation-induced epigenetic changes and Foxp3 expression are independent and complementary events required for Treg cell development. Immunity 37, 785-799. https://doi.org/10.1016/j.immuni.2012.09.010
  28. Ohnmacht, C., Park, J.H., Cording, S., Wing, J.B., Atarashi, K., Obata, Y., Gaboriau-Routhiau, V., Marques, R., Dulauroy, S., Fedoseeva, M., et al. (2015). MUCOSAL IMMUNOLOGY. The microbiota regulates type 2 immunity through RORgammat(+) T cells. Science 349, 989-993. https://doi.org/10.1126/science.aac4263
  29. Piper, C., Pesenacker, A.M., Bending, D., Thirugnanabalan, B., Varsani, H., Wedderburn, L.R., and Nistala, K. (2014). T cell expression of granulocyte-macrophage colony-stimulating factor in juvenile arthritis is contingent upon Th17 plasticity. Arthritis Rheumatol. 66, 1955-1960. https://doi.org/10.1002/art.38647
  30. Polansky, J.K., Kretschmer, K., Freyer, J., Floess, S., Garbe, A., Baron, U., Olek, S., Hamann, A., von Boehmer, H., and Huehn, J. (2008). DNA methylation controls Foxp3 gene expression. Eur. J. Immunol. 38, 1654-1663. https://doi.org/10.1002/eji.200838105
  31. Sakaguchi, S., Vignali, D.A., Rudensky, A.Y., Niec, R.E., and Waldmann, H. (2013). The plasticity and stability of regulatory T cells. Nat. Rev. Immunol. 13, 461-467. https://doi.org/10.1038/nri3464
  32. Sasidharan Nair, V., Song, M.H., and Oh, K.I. (2016). Vitamin C Facilitates Demethylation of the Foxp3 Enhancer in a Tet- Dependent Manner. J. Immunol. 196, 2119-2131. https://doi.org/10.4049/jimmunol.1502352
  33. Sefik, E., Geva-Zatorsky, N., Oh, S., Konnikova, L., Zemmour, D., McGuire, A.M., Burzyn, D., Ortiz-Lopez, A., Lobera, M., Yang, J., et al. (2015). MUCOSAL IMMUNOLOGY. Individual intestinal symbionts induce a distinct population of RORgamma(+) regulatory T cells. Science 349, 993-997. https://doi.org/10.1126/science.aaa9420
  34. Toker, A., Engelbert, D., Garg, G., Polansky, J.K., Floess, S., Miyao, T., Baron, U., Duber, S., Geffers, R., Giehr, P., et al. (2013). Active demethylation of the Foxp3 locus leads to the generation of stable regulatory T cells within the thymus. J. Immunol. 190, 3180-3188. https://doi.org/10.4049/jimmunol.1203473
  35. Wang, Y., Su, M.A., and Wan, Y.Y. (2011). An essential role of the transcription factor GATA-3 for the function of regulatory T cells. Immunity 35, 337-348. https://doi.org/10.1016/j.immuni.2011.08.012
  36. Wang, L., Liu, Y., Beier, U.H., Han, R., Bhatti, T.R., Akimova, T., and Hancock, W.W. (2013). Foxp3+ T-regulatory cells require DNA methyltransferase 1 expression to prevent development of lethal autoimmunity. Blood 121, 3631-3639. https://doi.org/10.1182/blood-2012-08-451765
  37. Wieczorek, G., Asemissen, A., Model, F., Turbachova, I., Floess, S., Liebenberg, V., Baron, U., Stauch, D., Kotsch, K., Pratschke, J., et al. (2009). Quantitative DNA methylation analysis of FOXP3 as a new method for counting regulatory T cells in peripheral blood and solid tissue. Cancer Res. 69, 599-608. https://doi.org/10.1158/0008-5472.CAN-08-2361
  38. Yang, X.P., Ghoreschi, K., Steward-Tharp, S.M., Rodriguez-Canales, J., Zhu, J., Grainger, J.R., Hirahara, K., Sun, H.W., Wei, L., Vahedi, G., et al. (2011). Opposing regulation of the locus encoding IL-17 through direct, reciprocal actions of STAT3 and STAT5. Nat. Immunol. 12, 247-254. https://doi.org/10.1038/ni.1995
  39. Yang, R., Qu, C., Zhou, Y., Konkel, J.E., Shi, S., Liu, Y., Chen, C., Liu, S., Liu, D., Chen, Y., et al. (2015). Hydrogen Sulfide Promotes Tet1- and Tet2-Mediated Foxp3 Demethylation to Drive Regulatory T Cell Differentiation and Maintain Immune Homeostasis. Immunity 43, 251-263. https://doi.org/10.1016/j.immuni.2015.07.017
  40. Yue, X., Trifari, S., Aijo, T., Tsagaratou, A., Pastor, W.A., Zepeda-Martinez, J.A., Lio, C.W., Li, X., Huang, Y., Vijayanand, P., et al. (2016). Control of Foxp3 stability through modulation of TET activity. J. Exp. Med. 213, 377-397. https://doi.org/10.1084/jem.20151438
  41. Zheng, Y., Josefowicz, S., Chaudhry, A., Peng, X.P., Forbush, K., and Rudensky, A.Y. (2010). Role of conserved non-coding DNA elements in the Foxp3 gene in regulatory T-cell fate. Nature 463, 808-812. https://doi.org/10.1038/nature08750

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