ＣＡＰ１遺伝子変異による急性骨髄性白血病の発症や再発機序の解明

2014 Fiscal Year Annual Research Report

• Project/Area Number: 24591412
• Research Institution: Nippon Medical School
• Principal Investigator: 山口 博樹 日本医科大学, 医学部, 准教授 (90297937)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Keywords: CAP1 / 急性骨髄性白血病 / 遺伝子変異

Outline of Annual Research Achievements

本研究はCAP1変異を認めるAMLの臨床的特徴や、CAP1変異の白血病発症や再発の機序を明らかにすることを目的としている。
オリゴ合成にてwild typeのCAP1cDNAを合成し、AML症例で発見されたCAP1変異をwild type CAP1cDNAよりPCR法とmutagenesis法を基にクローニングした。そしてCAP1GFP発現AcGFP1ベクターでCAP1のwild typeとmutant typeを発現させたHela細胞、 HEK293T細胞、ヒト血球系GM‐CSF依存性細胞株のMO7e細胞やF-36P細胞を作製した。
CAP1のwild typeとmutant typeを発現させたそれぞれの細胞に対してStaurosporin(STS)やetoposideにてアポトーシスを誘導しCAP1の細胞内局在を免疫染色法と細胞分画ごとのwestern blottingで検討したところ、wild typeはアポトーシスを誘導で細胞質からミトコンドリアに局在が変化をしたが、mutant typeは細胞質から移動しなかった。
Hela細胞に対してwild type CAP1に対するshRNAを2か所に設定し、pRNA-U6.1/NeoにてshRNAを導入し、90%以上knockdownされたクローンを作成した。そしてHela細胞にwild type、mutant type、wild/mutant type(1:1)のCAP1-GFPを発現させ、STSにて処理を行いアポトーシスの誘導をした。wild typeと比較してROSの蓄積、cytochrome c放出のwestern blotting、Caspase-3/7の活性はwild/mutant type(1:1)で1/2以下、mutant typeで1/5以下しかアポトーシスが誘導されてないことが示された。

Research Products

• [Journal Article] nhibition of S100A6 induces graft versus leukemia effects in MLL/AF4-positive ALL in human PBMC-SCID mice. (2014)
  • Author(s): Tamai.H, Miyake K, Yamaguchi H, Shimada T, Dan K, Inokuchi K.
  • Journal Title: Bone Marrow Transplantation Volume: 49(5) Pages: 699-703
  • DOI: 10.1038/bmt.2014.18. Epub 2014 Mar 3.
  • Peer Reviewed
• [Journal Article] Granulocyte colony-stimulating factor combined regimen in cord blood transplantation for acute myeloid leukemia: a nationwide retrospective analysis in Japan. (2014)
  • Author(s): 1.Konuma T, Ooi J, Uchida N, Ogawa H, Ohashi K, Kanamori H, Aotsuka N, Onishi Y, Yamaguchi H, Kozai Y, Nagamura-Inoue T, Kato K, Suzuki R, Atsuta Y, Kato S, Asano S, Takahashi S.
  • Journal Title: Haematologica Volume: 99(12) Pages: e264-8
  • DOI: 10.3324/haematol.2014.114504. Epub 2014 Sep 12.
  • Peer Reviewed / Open Access
• [Journal Article] 急性骨髄性白血病とエピジェネティクス制御関連遺伝子変異 (2014)
  • Author(s): 脇田知志、山口博樹
  • Journal Title: 日本臨床 Volume: 72(6) Pages: 1026-1032
• [Presentation] The prognostic and clinical feature of HSCT patients with chemotherapy refractory acute myeloid leukemia (2015)
  • Author(s): Keiko Fukunaga, Hiroki Yamaguchi, Yusuke Fujiwara, Shunsuke Yui, Takeshi Ryotokuji, Tsuneaki Hirakawa, Masahiro Okabe, Satoshi Wakita, Hayato Tamai, Muneo Okamoto, Kazutaka Nakayama, Koiti Inokuchi
  • Organizer: 第37回 日本造血細胞移植学会総会
  • Place of Presentation: 神戸
  • Year and Date: 2015-03-05 – 2015-03-07
• [Presentation] Decision Analysis of Allogeneic Hematopoietic Stem Cell Transplantation Versus Chemotherapy in Cytogenetically Standard-Risk Acute Myeloid Leukemia in First Complete Remission: The Impact of FLT3-ITD Profile (2014)
  • Author(s): Saiko Kurosawa, Hiroki Yamaguchi, et al.
  • Organizer: 56rd American Society of Hematology
  • Place of Presentation: San Francisco, USA
  • Year and Date: 2014-12-06 – 2014-12-08
• [Presentation] The D816V c-kit mutation confers higher proliferation activity by JAK-STAT and Src family kinase pathways compared to N822K c-kit mutation in core-binding factor acute myeloid leukemia (2014)
  • Author(s): Ikuko Omori, Hiroki Yamaguchi, Tomoaki Kitano, Noriko Miyake, Koichi Miyake, Koiti Inokuchi
  • Organizer: 56rd American Society of Hematology
  • Place of Presentation: San Francisco, USA
  • Year and Date: 2014-12-06 – 2014-12-08
• [Presentation] The prognostic impact of cytogenetic and genetic abnormality for the achievement of the second complete remission in adult patients with acute myeloid leukemia (2014)
  • Author(s): Keiko Fukunaga, Hiroki Yamaguchi, Satoshi Wakita, Ikuko Omori, Shunsuke Yui, Takeshi Ryotokuji, Tsuneaki Hirakawa, Kunihito Arai, Tomoaki Kitano, Fumiko Kosaka, Koiti Inokuchi
  • Organizer: 第76回日本血液学会学術集会
  • Place of Presentation: 大阪
  • Year and Date: 2014-10-31 – 2014-11-02
• [Presentation] The clinical features of PTPN11 gene mutation in Japanese patients with de novo AML (2014)
  • Author(s): Shunsuke Yui, Hiroki Yamaguchi, et al
  • Organizer: 第76回日本血液学会学術集会
  • Place of Presentation: 大阪
  • Year and Date: 2014-10-31 – 2014-11-02
• [Presentation] The clinical features and prognostic impact of acute myeloid leukemia with Epigenetics Modifying Gene mutation (2014)
  • Author(s): Anna Kobayashi, Hiroki Yamaguchi, Kensuke Usuki, Yutaka Kobayashi, Kenji Tajika, Gomi Seiji, Makoto Watanabe, Kentaro Azuma, Yuko Sato, Takahiro Todoroki, Ikuko Omori, Fukunaga Keiko, Satoshi Wakita, Tsuneaki Hirakawa, Takeshi Ryotokuji, Kunihito Arai, Tomoaki Kitano1, Fumiko Kosaka, Hayato Tamai, Koiti Inokuchi
  • Organizer: 第76回日本血液学会学術集会
  • Place of Presentation: 大阪
  • Year and Date: 2014-10-31 – 2014-11-02
• [Presentation] The clinical features and prognostic impact of de novo acute myeloid leukemia with WT1mutation (2014)
  • Author(s): Makoto Watanabe, Hiroki Yamaguchi, Kensuke Usuki, Yutaka Kobayashi, Kenji Tajika, Seiji Gomi, Anna Kobayashi, Kentaro Azuma, Yuko Sato, Takahiro Todoroki, Ikuko Omori, Fukunaga Keiko, Satoshi Wakita, Tsuneaki Hirakawa, Takeshi Ryotokuji, Kunihito Arai, Tomoaki Kitano, Fumiko Kosaka, Hayato Tamai, Koiti Inokuchi
  • Organizer: 第76回日本血液学会学術集会
  • Place of Presentation: 大阪
  • Year and Date: 2014-10-31 – 2014-11-02
• [Presentation] THE DIFFERENCE OF RCSD1-ABL1- LEUKEMOGENICITY BETWEEN EXON3 OF RCSD1/EXON 4 OF ABL1 AND EXON 2 OF RCSD1/EXON 4 OF ABL1 (2014)
  • Author(s): Hayato Tamai , Koichi Miyake, Hiroki Yamaguchi, Kazuo Dan, Koiti Inokuchi
  • Organizer: 19th. European Hematology Association
  • Place of Presentation: Milan, Italy
  • Year and Date: 2014-06-12 – 2014-06-15
• [Presentation] The prognostic impact of cytogenetic and genetic abnormality for the achievement of the second complete remission in adult patients with acute myeloid leukemia after first relapse (2014)
  • Author(s): Keiko Fukunaga, Hiroki Yamaguchi, Satoshi Wakita, Ikuko Omori, Shunsuke Yui, Takeshi Ryotokuji, Tsuneaki Hirakawa, Kunihito Arai, Tomoaki Kitano, Fumiko Kosaka, Kazuo Dan, Koiti Inokuchi.
  • Organizer: 19th. European Hematology Association
  • Place of Presentation: Milan, Italy
  • Year and Date: 2014-06-12 – 2014-06-15
• [Presentation] The prognostic impact of cytogenetic and genetic abnormality for the achievement of the second complete remission in adult patients with acute myeloid leukemia (2014)
  • Author(s): Keiko Fukunaga, Hiroki Yamaguchi, Satoshi Wakita, Ikuko Omori, Shunsuke Yui, Takeshi Ryotokuji, Tsuneaki Hirakawa, Kunihito Arai, Tomoaki Kitano, Fumiko Kosaka, Kazuo Dan, Koiti Inokuchi
  • Organizer: The 6th JSH International Symposium
  • Place of Presentation: 浜松
  • Year and Date: 2014-05-23 – 2014-05-24