B細胞系列への運命決定を制御する転写ネットワークの解明

• Project Area: Molecular mechanisms of cell fate determination in the cells that undergo stepwise differentiation to multiple pathways
• Project/Area Number: 25118732
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: The Institute of Physical and Chemical Research
• Principal Investigator: 伊川 友活 独立行政法人理化学研究所, 統合生命医科学研究センター, 上級研究員 (60450392)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: B細胞分化 / 運命決定 / 転写因子 / エピジェネティクス / リンパ球

Outline of Annual Research Achievements

B細胞を含むすべての血液・免疫細胞は造血幹細胞から作られる。その過程で多能性の造血幹細胞は徐々に分化能が限定されていき、最終的にB細胞にしかなれない前駆細胞に運命決定される。E2A、EBF1、PAX5など様々な転写因子がB細胞への運命決定に関わっているが、詳細は明らかでない。特にこれらの転写因子同士がどのようなネットワークを形成し、B細胞への運命決定を制御しているのか不明である。申請者らは最近このB細胞への運命決定に必要な転写ネットワークを調べる新しい分化誘導系を開発した。
そこで本研究では、培養細胞の遺伝子発現を網羅的に解析し転写因子間ネットワークのモデルを構築することを目的とした。本年度は研究計画に基づき、多能前駆細胞株（iLS細胞）からB細胞系列へ分化誘導後の経時サンプルを用いて網羅的な遺伝子発現解析（RNA-seq）およびエピジェネティック解析（ChIP-seq）を行った。さらにこのデータを元にバイオインフォマティクス解析を行った。
RNA-seq解析から転写因子の発現パターンによっていくつかのクラスターに分けられることが明らかとなった。培養0.5~1時間で誘導される転写因子（Nfil3, Nr4aなど)、培養2～4時間で誘導される転写因子（Tead1, Sox9など）、更にその後に誘導される転写因子（Ebf1, Pax5など）が存在することが明らかとなった。すなわち、Ebf1やPax5などB細胞特異的転写因子が発現する以前に、転写因子群が一過的かつダイナミックに次々と誘導されることが明らかとなった。これらのいわば転写カスケードがB細胞への運命決定を制御していると考えられる。

Research Progress Status

26年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

26年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Gene regulation. Transcribed enhancers lead waves of coordinated transcription in transitioning mammalian cells (2015)
  • Author(s): Arner E, Daub CO, Vitting-Seerup K, Andersson R, Lilje B, Drabløs F, Lennartsson A, Rönnerblad M, Hrydziuszko O, Vitezic M, Freeman TC, Alhendi AM, Arner P, Axton R, Baillie JK, Beckhouse A, Bodega B, Briggs J, Brombacher F, Davis M, Detmar M, Ehrlund A, Endoh M, Eslami A, Fagiolini M, et al.
  • Journal Title: Science Volume: 347 Issue: 6225 Pages: 1010-1014
  • DOI: 10.1126/science.1259418
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] The transcription repressors Bach2 and Bach1 promote B cell development by repressing myeloid program. (2014)
  • Author(s): Itoh-Nakadai A, Hikota R, Muto A, Kometani K, Matsui-Watanabe M, Sato Y, Kobayashi M, Nakamura A, Miura Y, Yano Y, Tashiro S, Sun J, Ikawa T, Ochiai K, Kurosaki T, and Igarashi K.
  • Journal Title: Nature Immunol. Volume: 15 Issue: 12 Pages: 1171-1180
  • DOI: 10.1038/ni.3024
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Zinc transporter SLC39A10/ZIP10 controls humoral immunity by modulating B-cell receptor signal strength. (2014)
  • Author(s): Hojyo S., T. Miyai, H. Fujishiro, M. Kawamura, T. Yasuda, A. Hijikata, B-H Bin, T. Irie, J. Tanaka, T. Atsumi, M. Murakami, M. Nakayama, O. Ohara, S. Himeno, H. Yoshida, H. Koseki, T. Ikawa, K. Mishima, and T. Fukada.
  • Journal Title: Proc Natl Acad Sci USA Volume: 111 Issue: 32 Pages: 11786-11791
  • DOI: 10.1073/pnas.1323557111
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Zinc transporter SLC39A10/ZIP10 facilitates antiapoptotic signaling during early B-cell development (2014)
  • Author(s): Miyai T, Hojyo S, Ikawa T, Kawamura M, Irie T, Ogura H, Hijikata A, Bin BH, Yasuda T, Kitamura H, Nakayama M, Ohara O, Yoshida H, Koseki H, Mishima K, Fukada T
  • Journal Title: Proceedings of the National Academy of Sciences of the United States of America Volume: 111 Issue: 32 Pages: 11780-11785
  • DOI: 10.1073/pnas.1323549111
  • Peer Reviewed / Open Access
• [Journal Article] Genetic and epigenetic control of early lymphocyte development (2014)
  • Author(s): Ikawa T
  • Journal Title: Curr Top Microbiol Immunol. Volume: 381 Pages: 1-20
  • DOI: 10.1007/82_2014_370
  • ISBN: 9783319073941, 9783319073958
  • Open Access / Acknowledgement Compliant
• [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] A promoter-level mammalian expression atlas. (2014)
  • Author(s): Forrest, A.R.R., Kawaji, H., Rehli, M., Baillie, J.K, de Hoon, M.J.L., Haberle, V., Lassmann, T., Kulakovskiy, I.V., Lizio, M., Itoh, M., Andersson, R., Mungall, C. J.,et al.
  • Journal Title: Nature Volume: 507 Issue: 7493 Pages: 462-470
  • DOI: 10.1038/nature13182
  • Peer Reviewed / Open Access
• [Journal Article] An atlas of active enhancers across human cell types and tissues. (2014)
  • Author(s): Anderson R, Gebhard C, Miguel-Escalada I, Hoof I, Bornholdt J, Boyd M, Chen Y, Zhao X, Schmidl C, Suzuki T, Collaborators(261) Enomoto H, etc.
  • Journal Title: Nature Volume: 507 Issue: 7493 Pages: 455-461
  • DOI: 10.1038/nature12787
  • Peer Reviewed / Open Access
• [Journal Article] Loss of mTOR complex 1 induces developmental blockage in early T-lymphopoiesis and eradicates T-cell acute lymphoblastic leukemia cells. (2013)
  • Author(s): Hoshii T, Kasada A, Hatakeyama T, Ohtani M, Tadokoro Y, Naka K, Ikenoue T, Ikawa T, Kawamoto H, Fehling HJ, Araki K, Yamamura K, Matsuda S, Hirao A
  • Journal Title: Proc Natl Acad Sci U S A. Volume: 111 Issue: 10 Pages: 3805-3810
  • DOI: 10.1073/pnas.1320265111
  • Peer Reviewed
• [Journal Article] MicroRNA-126-mediated control of cell fate in B-cell myeloid progenitors as a potential alternative to transcriptional factors. (2013)
  • Author(s): Okuyama K, Ikawa T, Gentner B, Hozumi K, Harnprasopwat R, Lu J, Yamashita R, Ha D, Toyoshima T, Chanda B, Kawamata T, Yokoyama K, Wang S, Ando K, Lodish HF, Tojo A, Kawamoto H, Kotani A
  • Journal Title: Proc Natl Acad Sci U S A. Volume: 110 Issue: 33 Pages: 13410-13415
  • DOI: 10.1073/pnas.1220710110
  • Peer Reviewed
• [Presentation] Gene regulatory network orchestrating B cell fate specification (2015)
  • Author(s): Tomokatsu Ikawa
  • Organizer: RIKEN EPIGENETICS in Kobe
  • Place of Presentation: 神戸市
  • Year and Date: 2015-02-13
• [Presentation] Epigenetic maintenance of T cell identity by Polycomb-mediated suppression of Pax5 (2014)
  • Author(s): Tomokatsu Ikawa
  • Organizer: 日本免疫学会
  • Place of Presentation: 京都市
  • Year and Date: 2014-12-11
• [Presentation] T細胞系列への運命決定とエピジェネティクス (2014)
  • Author(s): 伊川 友活
  • Organizer: LEGENDセミナー
  • Place of Presentation: 京都市
  • Year and Date: 2014-12-09
  • Invited
• [Presentation] Maintenance of T cell identity by Polycomb group proteins (2014)
  • Author(s): Ikawa T
  • Organizer: France-Japan Immunology meeting
  • Place of Presentation: フランス カシス
  • Year and Date: 2014-10-23
  • Invited
• [Presentation] Genetic and epigenetic control of early lymphocyte development (2014)
  • Author(s): Ikawa T
  • Organizer: Max Planck Institute seminar
  • Place of Presentation: ドイツ フライブルグ
  • Year and Date: 2014-10-20
  • Invited
• [Presentation] Polycomb group proteins Ring1A/B are required for the maintenance of T cell fate (2014)
  • Author(s): Ikawa T
  • Organizer: CSHL meeting
  • Place of Presentation: アメリカ コールドスプリングハーバー
  • Year and Date: 2014-04-22
• [Presentation] 細胞分化の研究に魅せられて (2014)
  • Author(s): 伊川 友活
  • Organizer: 新学術領域「細胞運命制御」若手の会
  • Place of Presentation: 浜松市
  • Year and Date: 2014-04-22
  • Invited
• [Presentation] A genetic network during the T cell lineage specification. (2013)
  • Author(s): Tomokatsu Ikawa
  • Organizer: The 6th International Workshop of Kyoto T cell conference 2013.
  • Place of Presentation: 京都市
• [Presentation] Roles of Polycomb group proteins in the maintenance of T cell fate. (2013)
  • Author(s): Tomokatsu Ikawa
  • Organizer: 15th International Congress of Immunology.
  • Place of Presentation: イタリア ミラノ
  • Invited
• [Presentation] Epigenetic regulation in T cell lineage commitment. (2013)
  • Author(s): Tomokatsu Ikawa
  • Organizer: The 5th LJI & IMS-RCAI Joint Workshop.
  • Place of Presentation: 横浜市
• [Presentation] Maintenance of T cell identity by Polycomb group complexes. (2013)
  • Author(s): Tomokatsu Ikawa
  • Organizer: 日本免疫学会
  • Place of Presentation: 千葉市
• [Presentation] A novel system to clarify the gene regulatory networks in B cell lineage commitment. (2013)
  • Author(s): Miyai Tomohiro, Tomokatsu Ikawa
  • Organizer: 日本免疫学会
  • Place of Presentation: 千葉市