腸内共生菌によるエピゲノム修飾を介した粘膜免疫制御メカニズムの解明

• Project/Area Number: 13J02667
• Research Category: Grant-in-Aid for JSPS Fellows
• Allocation Type: Single-year Grants
• Section: 国内
• Research Field: Immunology
• Research Institution: Chiba University
• Principal Investigator: 尾畑 佑樹 千葉大学, 大学院医学研究院, 特別研究員(DC1)
• Project Period (FY): 2013 – 2014
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥2,000,000 (Direct Cost: ¥2,000,000); Fiscal Year 2014: ¥1,000,000 (Direct Cost: ¥1,000,000); Fiscal Year 2013: ¥1,000,000 (Direct Cost: ¥1,000,000)
• Keywords: 腸内細菌 / 制御性T細胞 / 炎症性腸疾患 / DNAメチル化

Outline of Annual Research Achievements

ヒトの消化管の粘膜表面には、およそ100兆個にもおよぶ腸内細菌が生息している。消化管に慢性炎症を引き起こす炎症性腸疾患は、腸内細菌に対して宿主免疫系が過剰に応答することで発症すると考えられている。大腸における制御性T細胞(Treg)は、腸内細菌に対する免疫応答の抑制に重要な役割を果たしている。しかしながら、機能的なTregが大腸において維持される仕組みは不明であった。
著者らは、大腸Tregにおいて腸内細菌の定着依存的に発現増加する遺伝子としてUhrf1を同定した。Uhrf1はDNAメチル化の維持に必須な役割を果たしており、DNAメチル化は遺伝子発現の抑制に寄与している。そこで、Uhrf1を欠損したTregと正常なTregにおいて、DNAメチローム解析(MeDP-seq : Methylated DNA-precipitation-sequencing)およびトランスクリプトーム解析を行い、Uhrf1の標的分子の探索を試みた。これらデータの統合解析を行った結果、Uhrf1は細胞周期制御因子であるCdkn1aの遠位プロモーター領域のDNAメチル化を促すことで、その発現を負に抑制していることが示唆された。T細胞特異的にUhrf1を欠損したマウス(CD4^<Cre>Uhrf1^<F/F>)よりT細胞を取得し、Tregを分化誘導すると、G1-S移行期において細胞周期が停止することで、細胞増殖に異常をきたすことが判明した。一方で、siRNAを用いてCdkn1aをノックダウンしたUhrf1欠損TregではG1期における細胞周期の停止が軽減され、S期の細胞の割合が増加した。さらに著者らは、Uhrf1とCdkn1aの二重欠損マウス(CD4^<Cre>Uhrf1^<F/F>Cdkn1a^<-/->)を作成し、大腸Tregの割合を比較した。CD4^<Cre>Uhrf1^<F/F>では、抹消誘導型のTregの割合が激減する。一方で、CD4^<Cre>Uhrf1^<F/F>Cdkn1a^<-/->では、この割合が完全ではないものの有意に改善された。これらin vitroおよびin vivoの結果より、Uhrf1はDNAメチル化を介してCdkn1aの発現をエピジェネティックに抑制することで、Tregの細胞増殖を維持していることが明らかとなった。

Research Progress Status

本研究課題は平成26年度が最終年度のため記入しない。

Strategy for Future Research Activity

本研究課題は平成26年度が最終年度のため記入しない。

Research Products

• [Journal Article] Unique Lamina Propria Stromal Cells Imprint the Functional Phenotype of Mucosal Dendritic Cells (2015)
  • Author(s): Ildefonso Vicente-Suarez, Alexander Larange, Colin Reardon, Michael Matho, Sonia Feau, Yunji Park, Yuuki Obata, Rebecca Gold, Yiran Wang-Zhu, Chris Lena, Dirk Zajonc, Stephen Schoenberger, Mitch Kronenberg and Hilde Cheroutre.
  • Journal Title: Mucosal Immunol. Volume: 8 Issue: 1 Pages: 141-151
  • DOI: 10.1038/mi.2014.51
  • Peer Reviewed
• [Journal Article] Visualization of the entire differentiation process of murine M cells : suppression of their maturation in cecal patches. (2015)
  • Author(s): Shunsuke Kimura, Megumi Yamakami-Kimura, Yuuki Obata, Koji Hase, Hiroshi Kitamura, Hiroshi Ohno and Toshihiko Iwanaga.
  • Journal Title: Mucosal Immunol. Volume: 8 Issue: 3 Pages: 650-660
  • DOI: 10.1038/mi.2014.99
  • Peer Reviewed
• [Journal Article] Epigenetic modifications of the immune system in health and disease (2015)
  • Author(s): Yuuki Obata, Yukihiro Furusawa and Koji Hase.
  • Journal Title: Immunology and Cell Biology Volume: 93 Issue: 3 Pages: 226-232
  • DOI: 10.1038/icb.2014.114
  • Peer Reviewed
• [Journal Article] Mucosal barriology: The molecular machinery and physiological significance of multiple epithelial barriers (2015)
  • Author(s): Yuuki Obata and Koji Hase
  • Journal Title: Inflammation and Regeneration Volume: 35 Issue: 1 Pages: 003-013
  • DOI: 10.2492/inflammregen.35.003
  • NAID: 130004943877
  • ISSN: 1880-8190, 1880-9693
  • Peer Reviewed
• [Journal Article] The epigenetic regulator Uhrf1 facilitates proliferation and maturation of colonic regulatory T cells (2014)
  • Author(s): Yuuki Obata*, Yukihiro Furusawa*, Takaho A. Endo, Jafar Sharif, Daisuke Takahashi, Koji Atarashi, Manabu Nakayama, Satoshi Onawa, Yumiko Fujimura, Masumi Takahashi, Tomokatsu Ikawa, Takeshi Otsubo, Yuki I. Kawamura, Taeko Dohi, Shoji Tajima, Osamu Ohara, Kenya Honda, Shohei Hori, Hiroshi Ohno, Haruhiko Koseki and Koji Hase. 【* : 筆頭筆者】
  • Journal Title: Nature Immunology Volume: 15 Issue: 6 Pages: 571-579
  • DOI: 10.1038/ni.2886
  • Peer Reviewed / Open Access
• [Journal Article] Epithelial-stromal interaction via Notch signaling is essential for the full maturation of gut-associated lymphoid tissues (2014)
  • Author(s): Yuuki Obata, Shunsuke Kimura, Gaku Nakato, Keito Iizuka, Yurika Miyagawa, Yutaka Nakamura, Yukihiro Furusawa, Machiko Sugiyama, Keiichiro Suzuki, Masashi Ebisawa, Yumiko Fujimura, Hisahiro Yoshida, Toshihiko Iwanaga, Koji Hase and Hiroshi Ohno.
  • Journal Title: EMBO Reports Volume: 12 Issue: 12 Pages: 1297-1304
  • DOI: 10.15252/embr.201438942
  • Peer Reviewed
• [Journal Article] Pitfalls in global normalization of ChIP-seq data in CD4+ T cells treated with butyrate : A possible solution strategy. (2014)
  • Author(s): Yukihiro Furusawa, Takaho A. Endo, Yuuki Obata, Osamu Ohara, Hiroshi Ohno and Koji Hase.
  • Journal Title: Genomics Data Volume: 2 Pages: 176-180
  • Peer Reviewed
• [Journal Article] Commensal microbiota regulates T cell fate decision in the gut (2014)
  • Author(s): Yukihiro Furusawa, Yuuki Obata and Koji Hase.
  • Journal Title: Seminars in Immunopathology Volume: 37 Issue: 1 Pages: 017-025
  • DOI: 10.1007/s00281-014-0455-3
  • Peer Reviewed
• [Journal Article] Commensal microbe-derived butyrate induces colonic regulatory T cells (2013)
  • Author(s): Yukihiro Furusawa, Yuuki Obata, Shinji Fukuda, Takaho A. Endo, Gaku Nakato, Daisuke Takahashi, Yumiko Nakanishi, Chikako Uetake, Keiko Kato, Tamotsu Kato, Masumi Takahashi, Noriko N. Fukuda, Shinnosuke Murakami, Eiji Miyauchi, Shingo Hino, Koji Atarashi, Satoshi Onawa, Yumiko Fujimura, Trevor Lockett, Julie M. Clarke, David L. Topping, Masaru Tomita, Shohei Hori, Osamu Ohara, Tatsuya Morita, Haruhiko Koseki, J un Kikuchi, Kenya Honda, Koji Hase and Hiroshi Ohno
  • Journal Title: Nature Volume: 506 Issue: 7480 Pages: 446-450
  • DOI: 10.1038/nature12721
  • Peer Reviewed
• [Journal Article] AP-1B-Mediated Protein Sorting Regulates Polarity and Proliferation of Intestinal Epithelial Cells in Mice (2013)
  • Author(s): Koji Hase, Fubito Nakatsu, Masumi Ohmae, Kahushi Sugihara, Noriko Shioda, Daisuke Takahashi, Yuuki Obata, Yukihiro Furusawa, Yumiko Fujimura, Taisuke Yamashita, Shinji Fukuda, Hiroshi Okamoto, Masahide Asano, Shigenobu Yonemura and Hiroshi Ohno.
  • Journal Title: Gastroenterology Volume: 145 Issue: 3 Pages: 625-635
  • DOI: 10.1053/j.gastro.2013.05.013
  • Peer Reviewed
• [Journal Article] Distinct roles for CXCR6^+ and CXCR6^- CD4^+ T cells in the pathogenesis of chronic colitis (2013)
  • Author(s): Yasushi Mandai, Dasisuke Takahashi, Koji Hase, Yuuki Obata, Yukihiro Furusawa, Masashi Ebisawa, Tomoo Nakagawa, Toru Sato, Tatsuro Katsuno, Yasushi Saito, Takeshi Shimaoka, Osamu Yokosuka, Kotaro Yokote and Hiroshi Ohno
  • Journal Title: PLoS ONE
  • Peer Reviewed
• [Presentation] Epigenetic regulation of colonic immune homeostasis by gut microbiota (2015)
  • Author(s): Yuuki Obata, Yukihiro Furusawa and Koji Hase.
  • Organizer: The 4th NIF (Network of lmmunology Frontiers) Winter School on Advanced lmmunology 2015
  • Place of Presentation: Singapore
  • Year and Date: 2015-01-18 – 2015-01-23
• [Presentation] DNA methylation machinery in regulatory T cells secures immunological homeostasis in the colon. (2014)
  • Author(s): Yuuki Obata, Yukihiro Furusawa, Shohei Hori, Dohi Taeko, Hiroshi Ohno, Haruhiko Koseki and Koji Hase
  • Organizer: 43^<rd> Annual meeting of the Japanese society for Immunology
  • Place of Presentation: 国立京都国際会館(京都府・京都市)
  • Year and Date: 2014-12-10 – 2014-12-12
• [Presentation] Intestinal microbiota regulates epithelial barrier and mucosal immunity. (2014)
  • Author(s): Yuuki Obata and Koji Hase.
  • Organizer: International Symposium on Epithelial barrier and Transport 2014
  • Place of Presentation: 立命館大学BKCローム記念館(滋賀県・草津市)
  • Year and Date: 2014-11-01 – 2014-11-02
  • Invited
• [Presentation] Epigenetic regulator Uhrf1 facilitates proliferation and maturation of colonic regulatory T cells (2014)
  • Author(s): Yuuki Obata, Yukihiro Furusawa and Koji Hase.
  • Organizer: The3^<rd>annual International Graduate School Stude nt Immunology Conference 2014(IGSIC2014)
  • Place of Presentation: Dallas, USA
  • Year and Date: 2014-09-11 – 2014-09-13
• [Presentation] Epigenetic regulation by Uhrf1 is critical for functional expansion of colonic regulatory T cells (2013)
  • Author(s): Yuuki Obata, Yukihiro Furusawa, Shohei Hori, Dohi Taeko, Hiroshi Ohno, Haruhiko Koseki and Koji Hase
  • Organizer: 42^<nd> Annual meeting of the Japanese society for Immunology
  • Place of Presentation: Chiba
• [Presentation] Commensal Microbiota Shapes The Gut Immune System Through Epigenetic Modifications. (2013)
  • Author(s): Yuukl Obata, Yukihiro Furusawa and Koji Hase.
  • Organizer: International student research forum 2013 (ISRF2013)
  • Place of Presentation: Gold coast, Australia.
• [Presentation] Intestinal epithelium-intrinsic Notch signaling maintains mucosal immune homeostasis by ensuring epithelial barrier functions. (2013)
  • Author(s): Yuuki Obata, Daisuke Takahashi, Yukihiro Furusawa, Shohei Hori, Hiroshi Ohno, Koji Hase
  • Organizer: 15^<th> International Congress of Immunology (ICI)
  • Place of Presentation: Milan, Italy.
• [Presentation] Commensal Microbiota Shapes The Gut Immune System Through Epigenetic Modifications (2013)
  • Author(s): Yukihiro Furusawa, Yuuki Obata, Jafar Sharif, Shohei Hori, Hiroshi Ohno, Haruhiko Koseki and Koji Hase
  • Organizer: 16^<th> International Congress of mucosal Immunology (ICMI)
  • Place of Presentation: Vancouver, Canada.
• [Presentation] Commensal Microbiota Shapes The Gut Immune System Through Epigenet is Modifications. (2013)
  • Author(s): Yuuki Obata, Yukihiro Furusawa Hiroshi Ohno, Haruhiko Koseki and Koji Hase
  • Organizer: Cell Symposia, Microbiome
  • Place of Presentation: Lisbon, Portugal
• [Book] 実験医学 生体バリアの破綻と疾患 (2015)
  • Author(s): 尾畑佑樹, 長谷耕二
  • Publisher: 羊土社
• [Book] 腸内細菌によるエピゲノム修飾を介した腸管免疫制御メカニズムの解明 (2014)
  • Author(s): 尾畑佑樹
  • Publisher: 千葉医学雑誌 90巻5号
• [Book] 炎症と免疫2014年7月号(Vol.22 no.4) (2014)
  • Author(s): 尾畑佑樹、中村有孝、古澤之裕、長谷耕二
  • Publisher: 先端医学社
• [Book] エピゲノム制御タンパク質Uhrf1は腸内細菌の定着にともなう制御性T細胞の増殖および機能的な成熟を保証することにより炎症応答を抑制する (2014)
  • Author(s): 尾畑佑樹、長谷耕二
  • Publisher: ライフサイエンス新着論文レビュー
  • Description: Online総説
• [Book] エピジェネティクスキーワード辞典 (2013)
  • Author(s): 長谷耕二、古澤之裕、尾畑佑樹
  • Publisher: 羊土社
• [Book] 月刊細胞「腸内細菌による宿主免疫誘導」 (2013)
  • Author(s): 尾畑佑樹、中村有孝、長谷耕二
  • Publisher: ニューサイエンス社
• [Book] 感染・炎症・免疫「上皮バリア機構の破綻と腸管炎症の発症機構」 (2013)
  • Author(s): 長谷耕二、尾畑佑樹
  • Publisher: 医薬の門社