The study of the prognostic factor search in pediatric acute myeloid leukemia using the next generation sequencing and MLPA method

2016 Fiscal Year Annual Research Report

• Project/Area Number: 26461599
• Research Institution: National Center for Child Health and Development
• Principal Investigator: 大木 健太郎 国立研究開発法人国立成育医療研究センター, 小児血液・腫瘍研究部, 室長 (50400966)
• Co-Investigator(Kenkyū-buntansha): 林 泰秀 群馬県衛生環境研究所, その他部局等, 研究員 (30238133) ; 朴 明子 群馬県衛生環境研究所, その他部局等, 研究員 (50450375) ; 外松 学 群馬県衛生環境研究所, その他部局等, 研究員 (70251113)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: 遺伝子 / 小児がん / 急性骨髄性白血病 / 次世代シーケンス解析 / MLPA解析

Outline of Annual Research Achievements

本研究の目的は、小児急性骨髄性白血病(AML)の発症や進展に関わる分子機構を解明し、治療の層別化や分子標的薬の創薬に結びつけることにより、予後の向上に結びつけることである。
補助事業期間中に我々は小児AML23症例で次世代シーケンサーによる全エクソン解析を行い、コヒーシン複合体（RAD21、SMC3）、BCORL1、ASXL2遺伝子に新規の変異を見出した。得られた複数の新規遺伝子異常について、日本小児白血病リンパ腫グループ(JPLSG)のAML-05臨床試験に登録された370例で標的深読み塩基配列解析を行った。また、小児AML47症例で全トランスクリプトーム解析を行うとともに、リアルタイムPCR法を用いてAML99研究における発現アレイ解析から抽出された複数の遺伝子の発現解析を行った。
上記解析の結果、予後不良例ではクロマチン修飾関連遺伝子等の異常による複雑なエピジェネティックな変化が起きていると想定され、平成28年度に小児AML64症例で、Infinium Methylation EPIC BeadChip arrayによる網羅的メチル化解析を行った。AML1-MTG8再構成やMLL遺伝子再構成などの治療層別化に関わる病型毎にメチル化状態は異なるクラスターに分けられ、複数の新規メチル化異常が認められた。今後、本研究で集積した大量の遺伝子データおよびJPLSGと群馬県立小児医療センターの白血病データベースによる臨床データの解析により、白血病の病型と関連のあるゲノムパターンの異常、メチル化の特徴を抽出し、遺伝子・ゲノム、エピゲノムの異常に基づいた、病型診断、予後予測が構築できる可能性について詳細な検討を行い、本研究で新たに見出された遺伝子異常について、機能解析を行うことにより、それらの腫瘍化との関連性のさらなる検証を行い、標的遺伝子の同定を試みる予定である。

Research Products

• [Journal Article] ZNF384-related fusion genes consist of a subgroup with a characteristic immunophenotype in childhood B-cell precursor acute lymphoblastic leukemia. (2017)
  • Author(s): Hirabayashi S, Ohki K, Nakabayashi K, Ichikawa H, Momozawa Y, Okamura K, Ogata-Kawata H, Sakamoto H, Kato M, Matsuda K, Yoshida T, Matsumoto K, Hata K, Kubo M, Matsubara Y, Fukushima T, Koh K, Manabe A, Ohara A, Kiyokawa N.
  • Journal Title: Haematologica. Volume: 102 Pages: 118-127
  • DOI: 10.3324/haematol.2016.151035
  • Peer Reviewed
• [Journal Article] Prognostic impact of specific molecular profiles in pediatric acute megakaryoblastic leukemia in non-Down syndrome. (2017)
  • Author(s): Hara Y, Shiba N, Ohki K, Tabuchi K, Yamato G, Park MJ, Tomizawa D, Kinoshita K, Shimada A, Arakawa H, Saito AM, Kiyokawa N, Tawa A, Horibe K, Taga T, Adachi S, Taki T, and Hayashi Y.
  • Journal Title: Genes, Chromosomes & Cancer Volume: 14 February 2017 Pages: in print
  • DOI: 10.1002/gcc.22444
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] ASXL2 mutations are frequently found in pediatric AML patients with t(8;21)/RUNX1-RUNX1T1 and associated with a better prognosis. (2017)
  • Author(s): Yamato G, Shiba N, Yoshida K, Shiraishi Y, Hara Y, Ohki K, Okubo J, Okuno H, Chiba K, Tanaka H, Kinoshita A, Moritake H, Kiyokawa N, Tomizawa D, Park MJ, Sotomatsu M, Taga T, Adachi S, Tawa A, Horibe K, Arakawa H, Miyano S, Ogawa S, Hayashi Y.
  • Journal Title: Genes, Chromosomes & Cancer. Volume: 14 February 2017 Pages: in print
  • DOI: 10.1002/gcc.22443
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Whole-exome sequencing reveals the spectrum of gene mutations and the clonal evolution patterns in paediatric acute myeloid leukaemia. (2016)
  • Author(s): Shiba N, Yoshida K, Shiraishi Y, Okuno Y, Yamato G, Hara Y, Nagata Y, Chiba K, Tanaka H, Terui K, Kato M, Park MJ, Ohki K, Shimada A, Takita J, Tomizawa D, Kudo K, Arakawa H, Adachi S, Taga T, Tawa A, Ito E, Horibe K, Sanada M, Miyano S, Ogawa S, Hayashi Y.
  • Journal Title: Br J Haematol. Volume: 175 Pages: 476-489
  • DOI: 10.1111/bjh.14247
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] High PRDM16 expression identifies a prognostic subgroup of pediatric acute myeloid leukaemia correlated to FLT3-ITD, KMT2A-PTD, and NUP98-NSD1: the results of the Japanese Paediatric Leukaemia/Lymphoma Study Group AML-05 trial. (2016)
  • Author(s): Shiba N, Ohki K, Kobayashi T, Hara Y, Yamato G, Tanoshima R, Ichikawa H, Tomizawa D, Park MJ, Shimada A, Sotomatsu M, Arakawa H, Horibe K, Adachi S, Taga T, Tawa A, Hayashi Y
  • Journal Title: Br J Haematol. Volume: 172 Pages: 581-591
  • DOI: 10.1111/bjh.13869
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] Incidence and clinical relevance of MEF2D- and ZNF384-relattsuyoshied fusion genes in childhood B-cell precursor acute lymphoblastic leukemia: TCCSG L0416/0616 study (2016)
  • Author(s): 大木健太郎, 平林真介, 加藤元博, 中林一彦, 秦健一郎, 梶原良介, 高橋浩之, 福島敬, 康勝好, 真部淳, 小原明, 清河信敬.
  • Organizer: 第58回日本小児血液・がん学会学術集会
  • Place of Presentation: 東京
  • Year and Date: 2016-12-15 – 2016-12-17
• [Presentation] Frequency and clinical features of RUNX1 mutations in pediatric acute myeloid leukemia patients. (2016)
  • Author(s): 大和玄季, 柴徳生, 吉田健一, 原勇介, 大木健太郎, 大久保淳, 朴明子, 外松学, 清河信敬, 富澤大輔, 多賀崇, 足立壮一, 多和昭雄, 堀部敬三, 荒川浩一, 小川誠司, 林泰秀
  • Organizer: 第58回日本小児血液・がん学会学術集会
  • Place of Presentation: 東京
  • Year and Date: 2016-12-15 – 2016-12-17
• [Presentation] Novel risk stratification of infant acute myeloid leukemia using MLL-rearrangements and CBFA2T3-GLIS2. (2016)
  • Author(s): 原勇介, 柴徳生, 大木健太郎, 大和玄季, 朴明子, 富澤大輔, 滝智彦, 木下明俊, 清河信敬, 荒川浩一, 多和昭雄, 堀部敬三, 多賀崇, 足立壮一, 林泰秀
  • Organizer: 第58回日本小児血液・がん学会学術集会
  • Place of Presentation: 東京
  • Year and Date: 2016-12-15 – 2016-12-17
• [Presentation] Genetic Abnormalities and Prognosis in Pediatric B-precursor Acute Lymphoblastic Leukemia without conventional genetic abnormalities; Tokyo Children’s Cancer Study Group. (2016)
  • Author(s): Ohki K, Hirabayashi S, Yaguchi A, Terada K, Ohkubo J, Shiba N, Kato M, Fukushima T, Koh K, Hayashi Y, Manabe A, Ohara A, Kiyokawa N.
  • Organizer: 58th ASH Annual Meeting and Exposition
  • Place of Presentation: San Diego, USA
  • Year and Date: 2016-12-03 – 2016-12-06
  • Int'l Joint Research
• [Presentation] Clusters of Childhood Mixed Phenotype Acute Leukemia Diagnosed By Nation-Wide Immunophenotyping in Japan. (2016)
  • Author(s): Deguchi T, Kiyokawa N, Ohki K, Hashii Y, Manabe A, Adachi S, Saito AM, Horibe K, Komada Y.
  • Organizer: 58th ASH Annual Meeting and Exposition
  • Place of Presentation: San Diego, USA
  • Year and Date: 2016-12-03 – 2016-12-06
  • Int'l Joint Research
• [Presentation] Retrospective Evaluation of Correlations Between Genetic Backgrounds and Stem Cell Transplantation for De Novo Pediatric Acute Myeloid Leukemia: A Study from the Japan Pediatric Leukemia/Lymphoma Study Group (JPLSG) AML-05 Clinical Trial. (2016)
  • Author(s): Ueno T, Yamato G, Shiba N, Hara Y, Ohki K, Ichikawa H, Park MJ, Shimada A, Sotomatsu M, Arakawa H, Horibe K, Adachi S, Taga T, Tawa A, Hayashi Y.
  • Organizer: 58th ASH Annual Meeting and Exposition
  • Place of Presentation: San Diego, USA
  • Year and Date: 2016-12-03 – 2016-12-06
  • Int'l Joint Research
• [Presentation] Identification of Two Distinct Poor Prognostic Subgroups Related to High Expression of BMP2 or PRDM16 in Pediatric AML. (2016)
  • Author(s): Hara Y, Shiba N, Yamato G, Okubo J, Ohki K, Park MJ, Sotomatsu M, Tomizawa D, Taki T, Kinoshita A, Kiyokawa N, Taga T, Arakawa H, Tawa A, Horibe K, Adachi S, Hayashi Y.
  • Organizer: 58th ASH Annual Meeting and Exposition
  • Place of Presentation: San Diego, USA
  • Year and Date: 2016-12-03 – 2016-12-06
  • Int'l Joint Research
• [Presentation] The Clinical Features and Prognostic Impact of PRDM16 gene Expression in Adult Acute Myeloid Leukemia. (2016)
  • Author(s): Yamato G, Yamaguchi H, Handa H, Shiba N, Wakita S, Inokuchi K, Hara Y, Ohki K, Okubo J, Sotomatsu M, Arakawa H, Hayashi Y.
  • Organizer: 58th ASH Annual Meeting and Exposition
  • Place of Presentation: San Diego, USA
  • Year and Date: 2016-12-03 – 2016-12-06
  • Int'l Joint Research
• [Presentation] Transcriptome Analysis Revealed the Entire Genetic Understanding of Pediatric Acute Myeloid Leukemia with a Normal Karyotype. (2016)
  • Author(s): Shiba N, Yashida K, Shiraishi Y, Hara Y, Yamato G, Kaburagi T, Sanada M, Ohki K, Park MJ, Tomizawa D, Chiba K, Tanaka H, Sotomatsu M, Arakawa H, Horibe K, Taga T, Adachi S, Tawa A, Miyano S, Ogawa S, Hayashi Y.
  • Organizer: 58th ASH Annual Meeting and Exposition
  • Place of Presentation: San Diego, USA
  • Year and Date: 2016-12-03 – 2016-12-06
  • Int'l Joint Research
• [Presentation] Transcriptome analysis of 92 pediatric acute myeloid leukemia patients. (2016)
  • Author(s): Norio Shiba, Kenichi Yoshida, Genki Yamato, Yusuke Hara, Masashi Sanada, Yuichi Shiraishi, Kentaro Ohki, Myoug-Ja Park, Taeko Kaburaki, Daisuke Tomizawa, Manabu Sotomatsu, Hirokazu Arakawa, Keizo Horibe, Takashi Taga, Souichi Adachi, Akio Tawa, Satoru Miyano, Seishi Ogawa, Yasuhida Hayashi
  • Organizer: The 78th Annual Meeting of the Japanese Society of Hematology
  • Place of Presentation: 横浜
  • Year and Date: 2016-10-13 – 2016-10-15
• [Presentation] Molecular features and their clinical impact in infant acute myeloid leukemia (2016)
  • Author(s): Yusuke Hara, Norio Shiba, Kentaro Ohki, Genki Yamato, Myoug-Ja Park, Daisuke Tomizawa, Tomohiko Taki, Akitoshi Kinoshita, Hirokazu Arakawa, Akio Tawa, Keizo Horibe, Takashi Taga, Souichi Adachi, Yasuhide Hayashi
  • Organizer: The 78th Annual Meeting of the Japanese Society of Hematology
  • Place of Presentation: 横浜
  • Year and Date: 2016-10-13 – 2016-10-15
• [Presentation] Clinical features of patients with ASXL1 and ASXL2 mutations in pediatric acute myeloid leukemia (2016)
  • Author(s): Genki Yamato, Norio Shiba, Kenichi Yoshida, Kentaro Ohki, Jun Okubo, Yusuke Hara, Akitoshi Kinoshita, Daisuke Tomizawa, Manabu Sotomatsu, Takashi Taga, Souichi Adachi, Akio Tawa, Keizo Horibe, Hirokazu Arakawa, Seishi Ogawa, Yasuhida Hayashi
  • Organizer: The 78th Annual Meeting of the Japanese Society of Hematology
  • Place of Presentation: 横浜
  • Year and Date: 2016-10-13 – 2016-10-15
• [Remarks] 国立研究開発法人 国立成育医療研究センターホームページ
  • URL: https://www.ncchd.go.jp