ダウン症候群に伴う急性巨核球性白血病の多段階発症の分子機構

• Project Area: Molecular mechanisms of cell fate determination in the cells that undergo stepwise differentiation to multiple pathways
• Project/Area Number: 23118501
• 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: Hirosaki University
• Principal Investigator: 伊藤 悦朗 弘前大学, 医学(系)研究科(研究院), 教授 (20168339)
• Project Period (FY): 2011-04-01 – 2013-03-31
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥13,650,000 (Direct Cost: ¥10,500,000、Indirect Cost: ¥3,150,000); Fiscal Year 2012: ¥6,890,000 (Direct Cost: ¥5,300,000、Indirect Cost: ¥1,590,000); Fiscal Year 2011: ¥6,760,000 (Direct Cost: ¥5,200,000、Indirect Cost: ¥1,560,000)
• Keywords: ダウン症候群 / TAM / 巨核球性白血病 / 次世代シークエンサー / 発現クローニング / レトロウイルス

Outline of Annual Research Achievements

ダウン症候群に伴う急性巨核球性白血病（ML-DS）発症の過程で起こる多段階の遺伝子変異を解明し、細胞分化における運命決定メカニズムを明らかにすることを目的として本年度は以下の研究を進めた。
寛解時の検体が揃っているダウン症候群のTAMとAMKLの2５組（TAM 15例、AMKL 14例）の検体を用いて、全エクソンシーケンス解析を行った。今回の解析に用いた検体の内、4例はTAMとAMKLの両時期の検体が揃っていた。同一患者の寛解時検体から得られたデータと比較することで腫瘍細胞特異的な変異を同定した。サンガーシーケンスあるいはデイープシーケンスで確認できた変異数は129個であった。GATA1以外の変異数は平均してTAMでは0.7個、AMKLでは4.8個とAMKLで多い傾向にあった。注目すべきことに、DNA修復や転写調整に関わる同一経路 (pathway X) の分子をコードする遺伝子の変異が64％の症例で繰り返し認められた。
全エクソン解析の結果を確認するために、109例（TAM 41例、AMKL 49例、非ダウン症のAMKL 19例）に患者サンプルを増やしてターゲット・デイープシーケンス解析を行った。その結果、55%のAMKLにpathway Xの遺伝子変異が検出され、変異に重複が見られなかった。一方、TAMでは変異が見つかったのは41例中1例 (約2％)のみで、非ダウン症のAMKLでは19例中2例（約10％）であった。Tyrosine kinases あるいはRas pathwayの遺伝子変異は、AMKLでは41%、非ダウン症の AMKLで32%と両者に有意な差が認められなかった。TAMでは、41例中5例（12％）に変異が検出された。エピジェノム制御に関わる遺伝子変異は、AMKLでも非ダウン症のAMKLでも約20％に認められたが、TAMではほとんど検出されなかった。

Research Progress Status

24年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

24年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Naturally occurring oncogenic GATA1 mutants with internal deletions in transient abnormal myelopoiesis in Down syndrome (2013)
  • Author(s): Toki T, Kanezaki R, Kobayashi E, Kaneko H, Suzuki M, Wang R, Terui K, Kanegane H, Maeda M, Endo M, Mizuochi T, Adachi S, Hayashi Y, Yamamoto M, Shimizu R, Ito E.
  • Journal Title: Blood Volume: 121(16) Issue: 16 Pages: 3181-3184
  • DOI: 10.1182/blood-2012-01-405746
  • Peer Reviewed
• [Journal Article] Duodenal follicular lymphoma lacks AID but expresses BACH2 and has memory B-cell characteristics (2013)
  • Author(s): Takata K, Sato Y, Nakamura N, Tokunaka M, Miki Y, Yukie Kikuti Y, Igarashi K, Ito E, Harigae H, Kato S, Hayashi E, Oka T, Hoshii Y, Tari A, Okada H, Mohamad AA, Maeda Y, Tanimoto M, Kinoshita T, Yoshino T
  • Journal Title: Mod Pathol Volume: 26(1) Issue: 1 Pages: 22-31
  • DOI: 10.1038/modpathol.2012.127
  • NAID: 130004505772
  • Peer Reviewed
• [Journal Article] Rabbit antithymocyte globulin and cyclosporine as first-line therapy for children with acquired aplastic anemia (2012)
  • Author(s): Takahashi Y, Muramatsu H, Sakata N, Hyakuna N, Hamamoto K, Kobayashi R, Ito E, Yagasaki H, Ohara A, Kikuchi A, Morimoto A, Yabe H, Kudo K, Watanabe K, Ohga S, Kojima S
  • Journal Title: Blood Volume: 121(5) Issue: 5 Pages: 862-3
  • DOI: 10.1182/blood-2012-11-465633
  • Peer Reviewed
• [Journal Article] NEMO in T cells of patients with X-1 linked anhidrotic ectodermal dysplasia with immunodeficiency. (2012)
  • Author(s): Kawai T, Ito E, Heike T et al.
  • Journal Title: Blood Volume: 119 Pages: 5458-5466
  • Peer Reviewed
• [Journal Article] Identification of TRIB1 R107L gain-of-function mutation in human acute megakaryocytic leukemia (2012)
  • Author(s): Yokoyama T, Toki T, Aoki Y, Kanezaki R, Park MJ, Kanno Y, Takahara T, Yamazaki Y, Ito E, Hayashi Y, Nakamura T.
  • Journal Title: Blood Volume: 119 Issue: 11 Pages: 2608-2611
  • DOI: 10.1182/blood-2010-12-324806
  • Peer Reviewed
• [Journal Article] A novel mutation of ribosomal protein s10 gene in a Japanese patient with diamond-blackfan anemia (2012)
  • Author(s): Yazaki M, Kamei M, Ito Y, Konno Y, Wang R, Toki T, Ito E
  • Journal Title: J Pediatr Hematol Oncol Volume: 34(4) Issue: 4 Pages: 293-5
  • DOI: 10.1097/mph.0b013e31824a20ab
  • Peer Reviewed
• [Journal Article] Frequent somatic mosaicism of NEMO in T cells of patients with X-1 linked anhidrotic ectodermal dysplasia with immunodeficiency (2012)
  • Author(s): Kawai T, Ito E, 他21名
  • Journal Title: Blood Volume: (In press)
  • Peer Reviewed
• [Journal Article] Identification of TRIB1 R107L gain-of-function mutation in human acute megakaryocytic leukemia (2012)
  • Author(s): Yokoyama T, Toki T, Ito E, 他7名
  • Journal Title: Blood Volume: 119(11) Pages: 2608-2611
  • Peer Reviewed
• [Journal Article] Extensive gene deletions in Japanese patients with Diamond-Blackfan anemia (2012)
  • Author(s): Kuramitsu M, Toki T, Ito E, 他22名
  • Journal Title: Blood Volume: 119(10) Pages: 2376-84
  • Peer Reviewed
• [Journal Article] Extensive gene deletions in Japanese patients with Diamond-Blackfan anemia (2011)
  • Author(s): Kuramitsu M, Matsubara A, Morio T, Takagi M, Toki T, Terui K, RuNan W, Kanno H, Ohga S, Ohara A, Kitoh T, Sugita K, Kudo K, Matsubayashi T, Mizue N, Ozeki M, Masumi A, Momose H, Takizawa K, Mizukami T, Yamaguchi K, Ogawa S, Ito E, Hamaguchi I
  • Journal Title: Blood Volume: (in press) Issue: 10 Pages: 2376-84
  • DOI: 10.1182/blood-2011-07-368662
  • Peer Reviewed
• [Journal Article] Japan Cord Blood Bank Network. Cord blood transplantation from unrelated donors for children with acute lymphoblastic leukemia in Japan : the impact of methotrexate on clinical outcomes (2011)
  • Author(s): Kato K, Ito E, 他12名
  • Journal Title: Biol Blood Marrow Transplant Volume: 17(12) Pages: 1814-21
  • Peer Reviewed
• [Journal Article] Relapse of aplastic anemia in children after immunosuppressive therapy : a report from the Japan Childhood Aplastic Anemia Study Group (2011)
  • Author(s): Kamio T, Ito E, 他18名
  • Journal Title: Haematologica Volume: 96(6) Pages: 814-819
  • Peer Reviewed
• [Journal Article] CD7-positive acute myelomonocytic leukemia with trisomy 21 as a sole acquired chromosomal abnormality in two adolescents (2011)
  • Author(s): Kudo K, Ito E, 他4名
  • Journal Title: Leuk Res Volume: 35(9)
  • Peer Reviewed
• [Presentation] Genetic Basis of Myeloid Proliferation Relatd to Down syndrome (2012)
  • Author(s): Yoshida K, Toki T, Ito E, Ogawa S et al.
  • Organizer: 第54回アメリカ血液学会
  • Place of Presentation: 米国・アトランタ
  • Year and Date: 2012-12-08
• [Presentation] Comparison of Clinical Outcome Between Children with Aplastic Anemia and Refractory Cytopenia of Childhood Who Received Immunosuppressive Therapy with Antithymocyte Globulin and Cyclosporine (2011)
  • Author(s): Hama A, Ito E, 他13名
  • Organizer: 全米血液学会
  • Place of Presentation: San Diego, CA, USA
  • Year and Date: 2011-12-11
• [Presentation] GATA1 mutants lacking Rb-binding motif observed in transient abnormal myelopoiesis in Down syndrome (2011)
  • Author(s): Toki T, Ito E, 他12名
  • Organizer: 全米血液学会
  • Place of Presentation: San Diego, CA, USA
  • Year and Date: 2011-12-10
• [Presentation] Lower incidence of CRLF2 rearrangements in Down syndrome-associated acute lymphoblastic leukemia in Japan (2011)
  • Author(s): Hanada I, Toki T, Ito E, 他12名
  • Organizer: 全米血液学会
  • Place of Presentation: San Diego, CA, USA
  • Year and Date: 2011-12-10
• [Presentation] 先天性赤芽球瘻(Diamond-Blackfan貧血)の効果的診断法の確立関する研究 (2011)
  • Author(s): 伊藤悦朗, 照井君典, 土岐力, 他8名
  • Organizer: 第53回日本小児血液・がん学会学術集会
  • Place of Presentation: 前橋