骨髄造血幹細胞ニッチの構造と機能の解明

2013 Fiscal Year Annual Research Report

• Project/Area Number: 24249056
• Research Institution: The University of Tokyo
• Principal Investigator: 中内 啓光 東京大学, 医科学研究所, 教授 (40175485)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Keywords: 骨髄ニッチ / TGF-beta / 休眠状態

Research Abstract

1） 骨髄中の造血幹細胞ニッチの構造解析
申請者等は不活性化型TGF-betaが活性化できうるGFAP陽性細胞を中心としてヒト骨髄中における造血幹細胞ニッチの同定を行った。具体的には、ヒト造血生検から組織切片を作製し、造血に重要な因子であるSDF-1,SCF,TPO, Ang-1さらには、VE-cadherin, Osteopontinに対して反応する抗体を用い免疫染色を行った。結果、GFAP陽性細胞の周囲にはSDF-1,SCF,Ang-1陽性細胞が存在し、さらには血管の近傍にその構造が構築されていることが観察された。また、面白いことに、マウスとは異なる細胞がSDF-1やSCFを分泌していることが明らかとなった。これら結果は、GFAPを含むユニークな細胞集団がヒト造血幹細胞を維持する骨髄ニッチであるのではないかと示唆された。
2） 骨髄中の造血ニッチの関連機能分子の同定
ES細胞やiPS細胞を免疫不全マウスに移植することによりテラトーマが形成されるが、そのテラトーマ内には造血幹細胞が浸潤し骨髄ニッチ様の構造があると考えられた。申請者は様々な遺伝子をiPS細胞に導入し、より効率的にテラトーマ内に造血幹細胞がホーミングする遺伝子のスクリーニングを行った。現在、遺伝子導入iPS細胞を作製し免疫不全マウスに移植しており、今後、造血幹細胞のホーミングや増殖などの表現系する予定である。3） 造血幹細胞の細胞周期と冬眠を制御するTGF-bシグナリング機構の解明
TGF-b受容体は１型と２型が存在するが、造血幹細胞においては１型が発現していない。しかし、造血幹細胞においてTGF-bシグナルは重要な役割を果たしている。現在、造血幹細胞のcDNAライブラリーを作製しウイルスベクター系に移行中である。さらに、shRNAライブラリーにおいてもTGF-bシグナルが抑制する分子のスクリーニングを同時に進行しており、今後の結果が期待される。

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

当初の目標は８割は達成しており、将来に続く研究成果となっている。

Strategy for Future Research Activity

現在に至るまでに、マウスだけではなく、ヒトにおける造血幹細胞ニッチを理解することに成功している。今後はより詳細なメカニズムを明らかにすると共に、疾患を対象としたニッチの特性を利用した治療方法にもアプローチしていきたい。

Research Products

• [Journal Article] Nov/CCN3 regulates long-term repopulating activity of murine hematopoietic stem cells via integrin αvβ3. (2014)
  • Author(s): Ishihara J, Umemoto T, Yamato M, Shiratsuchi Y, Takaki S, Petrich BG, Nakauchi H, Eto K, Kitamura T, Okano T.
  • Journal Title: Int J Hematol. Volume: 99 Pages: 393-406
  • DOI: 10.1007/s12185-014-1534-x. Epub 2014 Feb 22.
  • Peer Reviewed
• [Journal Article] Haploinsufficiency of Sf3b1 leads to compromised stem cell function but not to myelodysplasia. (2014)
  • Author(s): Matsunawa M, Yamamoto R, Sanada M, Sato-Otsubo A, Shiozawa Y, Otsu M, Isono K, Koseki H, Nakauchi H, Ogawa S.
  • Journal Title: Leukemia. Volume: 73 Pages: 1-7
  • DOI: 10.1038/leu.2014.73.
  • Peer Reviewed
• [Journal Article] Expandable Megakaryocyte Cell Lines Enable Clinically Applicable Generation of Platelets from Human Induced Pluripotent Stem Cells. (2014)
  • Author(s): Nakamura S, Takayama N, Hirata S, Seo H, Endo H, Ochi K, Fujita KI, Koike T, Harimoto KI, Dohda T, Watanabe A, Okita K, Takahashi N, Sawaguchi A, Yamanaka S, Nakauchi H, Nishimura S, Eto K.
  • Journal Title: Cell Stem Cell. Volume: 14 Pages: 535-48
  • DOI: 10.1016/j.stem.2014.01.011.
  • Peer Reviewed
• [Journal Article] The TIF1β-HP1 System Maintains Transcriptional Integrity of Hematopoietic Stem Cells. (2014)
  • Author(s): Miyagi S, Koide S, Saraya A, Wendt GR, Oshima M, Konuma T, Yamazaki S, Mochizuki-Kashio M, Nakajima-Takagi Y, Wang C, Chiba T, Kitabayashi I, Nakauchi H, Iwama A.
  • Journal Title: Stem Cell Reports. Volume: 2 Pages: 145-52
  • DOI: 10.1016/j.stemcr.2013.12.008. eCollection 2014.
  • Peer Reviewed
• [Journal Article] Stage-specific roles for Cxcr4 signaling in murine hematopoietic stem/progenitor cells in the process of bone marrow repopulation. (2014)
  • Author(s): Lai CY, Yamazaki S, Okabe M, Suzuki S, Maeyama Y, Iimura Y, Onodera M, Kakuta S, Iwakura Y, Nojima M, Otsu M, Nakauchi H.
  • Journal Title: Stem Cells. Volume: - Pages: -
  • DOI: 10.1002/stem.1670.
  • Peer Reviewed
• [Journal Article] Homeodomain transcription factor meis1 is a critical regulator of adult bone marrow hematopoiesis. (2014)
  • Author(s): Ariki R, Morikawa S, Mabuchi Y, Suzuki S, Nakatake M, Yoshioka K, Hidano S, Nakauchi H, Matsuzaki Y, Nakamura T, Goitsuka R.
  • Journal Title: PLoS One. Volume: 9 Pages: e87646
  • DOI: 10.1371/journal.pone.0087646. eCollection 2014.
  • Peer Reviewed
• [Journal Article] Enzyme augmentation therapy enhances the therapeutic efficacy of bone marrow transplantation in mucopolysaccharidosis type II mice. (2014)
  • Author(s): Akiyama K, Shimada Y, Higuchi T, Ohtsu M, Nakauchi H, Kobayashi H, Fukuda T, Ida H, Eto Y, Crawford BE, Brown JR, Ohashi T.
  • Journal Title: Mol Genet Metab. Volume: 111 Pages: 139-46
  • DOI: 10.1016/j.ymgme.2013.09.013. Epub 2013 Sep 21.
  • Peer Reviewed
• [Journal Article] Derivation of Embryonic Stem Cell Lines from Parthenogenetically Developing Rat Blastocysts. (2014)
  • Author(s): Hirabayashi M, Goto T, Tamura C, Sanbo M, Hara H, Kato-Itoh M, Sato H, Kobayashi T, Nakauchi H, Hochi S.
  • Journal Title: Stem Cells Dev. Volume: 23 Pages: 107-14
  • DOI: 10.1089/scd.2013.0200. Epub 2013 Nov 7.
  • Peer Reviewed
• [Journal Article] Immortalization of Erythroblasts by c-MYC and BCL-XL Enables Large-Scale Erythrocyte Production from Human Pluripotent Stem Cells. (2013)
  • Author(s): Hirose S, Takayama N, Nakamura S, Nagasawa K, Ochi K, Hirata S, Yamazaki S, Yamaguchi T, Otsu M, Sano S, Takahashi N, Sawaguchi A, Ito M, Kato T, Nakauchi H, Eto K.
  • Journal Title: Stem Cell Reports. Volume: 1 Pages: 499-508
  • DOI: 10.1016/j.stemcr.2013.10.010.
  • Peer Reviewed
• [Journal Article] Profiling of MicroRNA in Human and Mouse ES and iPS Cells Reveals Overlapping but Distinct MicroRNA Expression Patterns. (2013)
  • Author(s): Razak SR, Ueno K, Takayama N, Nariai N, Nagasaki M, Saito R, Koso H, Lai CY, Murakami M, Tsuji K, Michiue T, Nakauchi H, Otsu M, Watanabe S.
  • Journal Title: PLoS One. Volume: 8 Pages: e73532.
  • DOI: 10.1371/journal.pone.0073532.
  • Peer Reviewed
• [Journal Article] Top-down motif engineering of a cytokine receptor for directing ex vivo expansion of hematopoietic stem cells. (2013)
  • Author(s): Saka K, Kawahara M, Teng J, Otsu M, Nakauchi H, Nagamune T.
  • Journal Title: J Biotechnol. Volume: 168 Pages: 659-65
  • DOI: 10.1016/j.jbiotec.2013.09.012. Epub 2013 Sep 23.
  • Peer Reviewed
• [Journal Article] Generation of Col2a1-EGFP iPS Cells for Monitoring Chondrogenic Differentiation. (2013)
  • Author(s): Saito T, Yano F, Mori D, Ohba S, Hojo H, Otsu M, Eto K, Nakauchi H, Tanaka S, Chung UI, Kawaguchi H.
  • Journal Title: PLoS One. Volume: 8 Pages: e74137.
  • DOI: 10.1371/journal.pone.0074137. eCollection 2013.
  • Peer Reviewed
• [Journal Article] Do pluripotent stem cells exist in adult mice as very small embryonic stem cells? (2013)
  • Author(s): Miyanishi M, Mori Y, Seita J, Chen JY, Karten S, Chan CK, Nakauchi H, Weissman IL.
  • Journal Title: Stem Cell Reports. Volume: 1 Pages: 198-208
  • DOI: 10.1016/j.stemcr.2013.07.001.
  • Peer Reviewed
• [Journal Article] Clonal analysis unveils self-renewing lineage-restricted progenitors generated directly from hematopoietic stem cells. (2013)
  • Author(s): Yamamoto R, Morita Y, Ooehara J, Hamanaka S, Onodera M, Rudolph KL, Ema H, Nakauchi H.
  • Journal Title: Cell. Volume: 154 Pages: 1112-26.
  • DOI: 10.1016/j.cell.2013.08.007.
  • Peer Reviewed
• [Journal Article] Recurrent mutations in multiple components of the cohesin complex in myeloid neoplasms. (2013)
  • Author(s): Kon A, Shih LY, Minamino M, Sanada M, Nakauchi H. et al.
  • Journal Title: Nature Genet. Volume: 45 Pages: 1232-7
  • DOI: 10.1038/ng.2731. Epub 2013 Aug 18.
  • Peer Reviewed
• [Journal Article] An in vitro expansion system for generation of human iPS cell-derived hepatic progenitor-like cells exhibiting a bipotent differentiation potential. (2013)
  • Author(s): Yanagida A, Ito K, Chikada H, Nakauchi H, Kamiya A.
  • Journal Title: PLoS One. Volume: 8 Pages: e67541.
  • DOI: 10.1371/journal.pone.0067541. Print 2013.
  • Peer Reviewed
• [Journal Article] CAMT-iPS Cells Exhibiting Defective MPL Signaling Dysregulate Megakaryopoiesis and Erythropoiesis. (2013)
  • Author(s): Shinji Hirata, Naoya Takayama,Ryoko Jono-Ohnishi, Hiroshi Endo, Sou Nakamura, Takeaki Dohda, Masanori Nishi, Yuhei Hamazaki, Ei-ichi Ishii, Shin Kaneko, Makoto Otsu, Hiromitsu Nakauchi, Shinji Kunishima, and Koji Eto.
  • Journal Title: J Clin Invest. Volume: 123 Pages: 3802-14.
  • DOI: 10.1172/JCI64721.
  • Peer Reviewed
• [Journal Article] Two differential flows in a bioreactor promoted platelet generation from human pluripotent stem cell-derived megakaryocytes. (2013)
  • Author(s): Nakagawa Y, Nakamura S, Nakajima M, Endo H, Dohda T, Takayama N, Nakauchi H, Arai F, Fukuda T, Eto K.
  • Journal Title: Exp Hematology. Volume: 41 Pages: 742-8
  • DOI: 10.1016/j.exphem.2013.04.007. Epub 2013 Apr 22.
  • Peer Reviewed
• [Journal Article] Generation of engraftable hematopoietic stem cells from induced pluripotent stem cells by way of teratoma formation. (2013)
  • Author(s): Suzuki N, Yamazaki S, Yamaguchi T, Okabe M, Masaki H, Takaki S, Otsu M, Nakauchi H.
  • Journal Title: Mol Ther. Volume: 21 Pages: 1424-31
  • DOI: 10.1038/mt.2013.71. Epub 2013 May 14
  • Peer Reviewed
• [Journal Article] Generation of transgenic mouse line expressing Kusabira Orange throughout body, including erythrocytes, by random segregation of provirus method. (2013)
  • Author(s): Hamanaka S, Ooehara J, Morita Y, Ema H, Takahashi S, Miyawaki A, Otsu M, Yamaguchi T, Onodera M, Nakauchi H.
  • Journal Title: Biochem Biophys Res Commun. Volume: 435 Pages: 586-591
  • DOI: 10.1016/j.bbrc.2013.05.017. Epub 2013 May 17.
  • Peer Reviewed
• [Presentation] "Novel non-stepwise early differentiation pathways of HSCs revealed by 5-lineage in vivo tracing" (2013)
  • Author(s): Hiromitsu Nakauchi
  • Organizer: Gordon Research Conference “Stem Cells & Cancer”
  • Place of Presentation: Les Diablerets, Switzerland
  • Year and Date: 20130421-20130426
  • Invited
• [Presentation] Exploring heterogeneity and hierarchy within the most primitive hematopoietic stem cell compartment
  • Author(s): 中内啓光
  • Organizer: 日本分子生物学会シンポジウム
  • Place of Presentation: 兵庫
  • Invited
• [Book] 幹細胞研究と再生医療 (2013)
  • Author(s): 中内啓光（編集）
  • Total Pages: 238
  • Publisher: 南山堂
• [Book] 直伝！フローサイトメトリー (2013)
  • Author(s): 中内啓光（監修）清田純（編集）
  • Total Pages: 278
  • Publisher: 羊土社