マウスES細胞を材料とした人工胸腺の開発

• Project/Area Number: 20060035
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: The Institute of Physical and Chemical Research
• Principal Investigator: 河本 宏 The Institute of Physical and Chemical Research, 免疫発生研究チーム, チームリーダー (00343228)
• Project Period (FY): 2008 – 2009
• Project Status: Completed (Fiscal Year 2009)
• Budget Amount: ¥10,000,000 (Direct Cost: ¥10,000,000); Fiscal Year 2009: ¥5,000,000 (Direct Cost: ¥5,000,000); Fiscal Year 2008: ¥5,000,000 (Direct Cost: ¥5,000,000)
• Keywords: 人工胸腺 / ES細胞 / Foxn1 / 内胚葉 / 胸腺上皮細胞

Research Abstract

本研究は、免疫不全患者に対し、人工胸腺を構築することによりT細胞の再生を促進するという治療戦略の基盤となる技術の開発を目的とする。人工胸腺のもとになる細胞としてES細胞を用いる。20年度は、胸腺上皮細胞が誘導されるとGFP陽性になるES細胞(Foxn1-Cre-Lox-GFP-ES細胞)を用いて、内胚葉系細胞の誘導を誘導した上でWnt、 BMP4などを添加して分化誘導をかけてみたが、GFP陽性細胞の誘導にはいたらなかった。そこで21年度は、Foxn1プロモーター下に直接GFPを発現するES細胞を作製した。このES細胞はThomas Boehm(Max-Planck Institute of Immunobiology、ドイツ)より入手したFoxn1-GFPマウスから樹立したものである。この細胞を主に用いてさらなる条件検討を加えることを試みた。前年度と同じくWnt、 BMP4などを添加した他、retinoic acidなども球したが、残念ながらGFP陽性細胞の誘導にはいたらなかった。しかし、平行して行った胸腺上皮細胞の分化増殖シグナルを検索する研究においては、大変重要な知見がえられた。この研究では、Foxn1-Creを用いて胸腺上皮特異的にstat3を欠損させたマウスを作成した。すなわち、Foxn1-Creとstat3-flox/floxマウスをかけあわせた。Foxn1-Cre:stat3-flox/floxマウスでは出生後の胸腺において胸腺皮質は正常であったが胸腺髄質が著明な低形成を呈することを見いだした。一方、新生児期のマウスの髄質は正常に形成されていた。これは、stat3を介するシグナル経路が出生後の胸腺髄質の発達に必要であることを示している。この知見は胸腺上皮細胞の分化誘導系の開発において重要な情報である。

Research Products

• [Journal Article] Thymic cysts originate from Foxn1 positive thymic medullary epithelium (2010)
  • Author(s): Vroegindeweij, E., S Crobach, M Itoic, R Satoh, S Zuklys, C Happee, W TV Germeraad, J J Cornelissen, T Cupedo, G A Hollander, H Kawamoto, W van Ewijk
  • Journal Title: Mol Immunol 47 Pages: 1106-113
  • Peer Reviewed
• [Journal Article] Generation of functional NKT cells in vitro from embryonic stem cells bearing rearranged invariant Vα14-Jα18 TCRα gene (2010)
  • Author(s): Watarai H, A Rybouchkin, N Hongo, Y Nagata, S Sakata, E Sekine, N Dashtsoodol, T Tashiro, S-I Fujii, K Shimizu, K Mori, K.Masuda, H Kawamoto, H Koseki, M Taniguchi
  • Journal Title: Blood 115 Pages: 230-237
  • Peer Reviewed
• [Journal Article] In vitro human G-CSF mobilized CD34+ derived T-cell progenitors fully mature in vivo (2010)
  • Author(s): Meek B, S Cloosen, C Borsotti, J Van Elssen, J Vanderlocht, M Schnijderberg, M van der Poel, B Leewis, R Hesselink, M G Manz, Y Katsura, H Kawamoto, W TV Germeraad, G MJ Bos
  • Journal Title: Blood 115 Pages: 261-264
  • Peer Reviewed
• [Journal Article] Innate production of Th2 cytokines by adipose tissue-associated c-Kit+Sca-1+ lymphocytes (2010)
  • Author(s): Moro K., T.Yamada, M.Tanabe, T.Takeuchi, T.Ikawa, H.Kawamoto J-I.Furusawa, M.Ohtani, H.Fujii, S.Koyasu
  • Journal Title: Nature 463 Pages: 540-544
  • Peer Reviewed
• [Journal Article] Direct hematological toxicity and illegitimate chromosomal recombination caused by the systemic activation of CreERT2 (2009)
  • Author(s): Higashi, A.Y., T.Ikawa, M.Muramatsu, A.N.Economides, A.Niwa, T.Okuda, A.J.Murphy, J.Rojas, T.Heike, T.Nakahata, H.Kawamoto, T.Kita, Motoko Yanagita
  • Journal Title: J.Immunol. 182 Pages: 5633-5640
  • Peer Reviewed
• [Journal Article] A novel gene essential for the development of single positive thymocytes (2009)
  • Author(s): Kakugawa K., T.Yasuda, I.Miura, A.Kobayashi, H.Fukiage, R.Satoh, M.Matsuda, H.Koseki, S.Wakana, H.Kawamoto, H.Yoshida
  • Journal Title: Mol.Cell.Biol 29 Pages: 5128-5135
  • Peer Reviewed
• [Journal Article] Ectopically expressed PIR-B on T cells constitutively binds to MHC class I and attenuates T helper type 1 responses (2009)
  • Author(s): Imada M, Masuda K, Satoh R, Ito Y, Goto Y, Matsuoka T, Endo S, Nakamura A, Kawamoto H, Takai T
  • Journal Title: Int.Immnol 21 Pages: 1151-1156
  • NAID: 10027389800
  • Peer Reviewed
• [Journal Article] Adult T cell progenitors retain myeloid potential (2009)
  • Author(s): Wada H, Masuda K, Satoh R, Kakugawa K, Ikawa, T, Katsura Y, Kawamoto H.
  • Journal Title: Nature 452 Pages: 768-772
  • Peer Reviewed
• [Journal Article] Lipid-mediated Presentation of MHC class II Molecules guides thymocytes to the CD4 lineage (2009)
  • Author(s): Komaniwa S, H Hayashi, H Kawamoto, T Ikawa, Satoshi B.Sato, Y Katsura, K Udaka.
  • Journal Title: Eur.J.Immunology 39 Pages: 96-122
  • Peer Reviewed
• [Journal Article] Cascading suppression of transcriptional silencers by ThPOK seals helper Tcell fate. (2008)
  • Author(s): Muroi, S., Y Naoe, C Miyamoto, K Akiyama, T Ikawa, K Masuda, H Kwamoto, I Taniuchi.
  • Journal Title: Nature Immonology 9 Pages: 1131-1121
  • Peer Reviewed
• [Journal Article] Id2 deficient mice develop myeloproliferative disorder whereas forced Id2 expression interferes with the development of BCR/ABL induced myeloid leukemia. (2008)
  • Author(s): Ko J., N Patel, T Ikawa, H Kwamoto, O Frank, R Rivera, RA van Etten, C Murre
  • Journal Title: PNAS 105 Pages: 12967-12972
  • Peer Reviewed
• [Journal Article] Aire contorols the differentiation of thymic epithelial cells in the medulla for the estabilishment of self-tolerance. (2008)
  • Author(s): Yano M, Kuroda N, Han H, Horike M, Nishikawa Y, Kiyonari H, Maemura K, Yanagawa Y, Obata K, Takahashi S, Ikawa T, Satoh R, Kawamoto H, Muroi Y, Matsumoto M.
  • Journal Title: JEM 205 Pages: 2827-2838
  • Peer Reviewed
• [Journal Article] Notch activation in thymic epithelial cells induces development of thymic microenvironments.
  • Author(s): Kyoko Masuda, Wilfred T.V.Germeraad, Rumi Satoh, Manami Itoi, Tomokatsu Ikawa, Nagahiro Minato, Yoshimoto Katsura, Willem van Ewijk, Hiroshi Kawamoto
  • Journal Title: Molecular Immunology (in press)
  • Peer Reviewed
• [Presentation] Lineage restriction pathway in hematopoiesis : Revision of the classical concept of myeloid-lymphoid dichotomy (2010)
  • Author(s): 河本宏
  • Organizer: Immunology Seminar Series
  • Place of Presentation: ハーバード大学医学部(Boston, USA)
  • Year and Date: 2010-02-24
• [Presentation] 血液細胞の新しい分化経路モデル-混合型白血病細胞の起源についての考察- (2009)
  • Author(s): 河本宏
  • Organizer: 近畿血液学地方会
  • Place of Presentation: メルパルク京都(京都)
  • Year and Date: 2009-12-12
• [Presentation] T cell lineage determination : termination of myeloid potential in T cell progenitors (2009)
  • Author(s): 河本宏
  • Organizer: 日本免疫学会シンポジウム
  • Place of Presentation: 大阪国際会議場(大阪)
  • Year and Date: 2009-12-02
• [Presentation] 血液細胞の新しい分化経路モデル-混合型白血病細胞の起源についての考察- (2009)
  • Author(s): 河本宏
  • Organizer: 小児血液学会
  • Place of Presentation: 東京ベイホテル東急(浦安)
  • Year and Date: 2009-11-27
• [Presentation] 血液細胞の新しい分化経路モデル-多様な細胞種は、どういう順序でつくられるか- (2009)
  • Author(s): 河本宏
  • Organizer: 数理生物学会
  • Place of Presentation: 東大駒場数理センター(東京)
  • Year and Date: 2009-09-09
• [Presentation] Induction of self-renewal program in developing hematopoietic progenitors by inhibiting their differentiation (2009)
  • Author(s): 河本宏
  • Organizer: JST-CIRM workshop
  • Place of Presentation: 京都フジタホテル(京都)
  • Year and Date: 2009-09-01
• [Presentation] リンパ球の起源と分化 (2009)
  • Author(s): 河本宏
  • Organizer: 比較免疫学会シンポジウム
  • Place of Presentation: 日本大学(藤沢)
  • Year and Date: 2009-08-05
• [Presentation] リンパ球から人工万能細胞(iPS細胞)をつくる (2009)
  • Author(s): 河本宏
  • Organizer: 動物細胞工学会大会
  • Place of Presentation: 筑波大学(つくば)
  • Year and Date: 2009-07-24
• [Presentation] Retention and termination of myeloid potential in T cell progenitors (2009)
  • Author(s): 河本宏
  • Organizer: Kyoto T cell Conference
  • Place of Presentation: 京大会館(京都)
  • Year and Date: 2009-06-02
• [Presentation] Myeloid potential is retained in T cell progenitors : Revised scheme for hematopoiesis (2009)
  • Author(s): Hiroshi Kawamoto
  • Organizer: LIAI-RCAI Joint Workshop2009
  • Place of Presentation: 横浜
  • Year and Date: 2009-01-07
• [Presentation] 造血モデルの見直し : ミエロイド基本型モデルの提唱 (2008)
  • Author(s): Hiroshi Kawamoto
  • Organizer: 分子生物学会ワークショップ
  • Place of Presentation: 神戸
  • Year and Date: 2008-12-13
• [Presentation] T cell progenitors retain myeloid potential : Revised scheme for hematopoiesis (2008)
  • Author(s): Hiroshi Kawamoto
  • Organizer: 日本免疫学会シンポジウム
  • Place of Presentation: 京都
  • Year and Date: 2008-12-02
• [Presentation] 造血モデルの見直し : ミエロイド基本型モデルの提唱 (2008)
  • Author(s): Hiroshi Kawamoto
  • Organizer: 血液科学セミナー
  • Place of Presentation: 東京
  • Year and Date: 2008-11-15
• [Presentation] T cell progenitors retain myeloid potential : Revised scheme for hematopoiesis (2008)
  • Author(s): Hiroshi Kawamoto
  • Organizer: ThymUS workshop on T cell biology
  • Place of Presentation: San Juan, Puerto-rico
  • Year and Date: 2008-11-10
• [Presentation] Adult T cell progenitors retain myeloid potential (2008)
  • Author(s): Hiroshi Kawamoto
  • Organizer: Cold Spring Harbor meeting on Gene Express ion & Signaling in the Immune System
  • Place of Presentation: Cold Spring Harbor
  • Year and Date: 2008-04-23