Integrated analyses including general epigenetic analysis in pediatric acute myeloid leukemia

• Project/Area Number: 20K08744
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 54010:Hematology and medical oncology-related
• Research Institution: Gunma Institute of Public Health and Environmental Sciences
• Principal Investigator: Hayashi Yasuhide 群馬県衛生環境研究所, 研究企画係, 研究員 (30238133)
• Co-Investigator(Kenkyū-buntansha): 外松 学 群馬県衛生環境研究所, 研究企画係, 研究員 (70251113) ; 河崎 裕英 群馬県衛生環境研究所, 研究企画係, 研究員 (80278621) ; 山田 佳之 群馬県衛生環境研究所, 研究企画係, 研究員 (80309252)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2022: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2021: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2020: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 小児白血病 / 次世代シーケンサー / 網羅的メチル化解析 / ７モノソミー / RAS関連遺伝子 / TP53遺伝子 / 予後因子 / ターゲット / 7モノソミー / RB1遺伝子 / ターゲットシーケンス / 急性骨髄性白血病 / 遺伝子 / マイクロアレイ / トランスクリプトーム / メチル化アレイ

Outline of Research at the Start

小児急性骨髄性白血病（AML）の次世代シーケンサーによるトランスクリプトーム解析、網羅的メチル化解析とAML関連343遺伝子のTarget シーケンス解析による統合的解析により、新規遺伝子をみいだし、性状を明らかにし、それを通じて白血病の発症と進展機序を明らかにする。特にメチル化で見出した層別化に関与する遺伝子を解析の症例を増やして検討し、小児ＡＭＬの予後の指標を見出して治療法の決定に貢献し、分子標的療法に通じる分子遺伝学的基盤を構築する。

Outline of Final Research Achievements

The prognosis of pediatric acute myeloid leukemia (AML) has improved via stratification therapy. RAS pathway alterations were detected in 80 (24.4%) of 328 patients: NF1 (n=7, 2.1%), PTPN11 (n=15, 4.6%), CBL (n=6, 1.8%), NRAS (n=44, 13.4%), KRAS (n=12, 3.7%). NF1 alterations were frequently detected in cases with complex karyotype (P=0.031) and were found to be independent predictors of poor overall survival in multivariate analysis (P=0.007). We detected recurrent internal tandem duplication in upstream binding transcription factor (UBTF-ITD) in 1.2% (6/503) of pediatric AML patients with de novo AML. Among AML cases with 7 monosomy, KMT2C was frequently found in 15 cases (4.5%) and associated with poor prognosis. Genome-wide DNA methylation patterns in 64 pediatric AML patients were categorized into 4 clusters. Our results suggest that DNA methylation levels at specific CpG sites are useful to support genetic alterations and gene expression patterns of AML cases.

Academic Significance and Societal Importance of the Research Achievements

難治性の小児急性骨髄性白血病（AML）の遺伝子研究で次世代シーケンスを用いてRAS関連遺伝子、TP53とRB1遺伝子、7番染色体上のKMT2C遺伝子等が予後因子であることを明らかにした。遺伝子の縦列重複を同定した遺伝子の中でUBTF-ITDが予後不良因子であることを見出した。さらに網羅的メチル化解析で高メチル化群は著しく予後不良であった。新たに予後と相関する遺伝子の多数の発見は小児AMLの治療の層別化と予後の改善に貢献するだけではなくて、がんの発生に根源的に関与すると思われる小児がんで見出したデータは成人のがんの発生の基盤となり、すべてのがんの研究と治療にも貢献すると思われた。

Research Products

• [Journal Article] UBTF-internal tandem duplication as a novel poor prognostic factor in pediatric acute myeloid leukemia. (2023)
  • Author(s): Kaburagi T, Shiba N, Yamato G, Yoshida K, Tabuchi K, Ohki K, Ishikita E, Hara Y, Shiraishi Y, Kawasaki H, Sotomatsu M, Takizawa T, Taki T, Kiyokawa N, Tomizawa D, Horibe K, Miyano S, Taga T, Adachi S, Ogawa S, Hayashi Y.
  • Journal Title: Genes Chromosomes Cancer Volume: 62 Issue: 4 Pages: 202-209
  • DOI: 10.1002/gcc.23110
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] TP53 and RB1 alterations characterize poor prognostic subgroups in pediatric acute myeloid leukemia. (2023)
  • Author(s): Hara Y, Shiba N, Yoshida K, Yamato G, Kaburagi T, Shiraishi Y, Ohki K, Shiozawa Y, Kawamura M, Kawasaki H, Sotomatsu M, Takizawa T, Matsuo H, Shimada A, Kiyokawa N, Tomizawa D, Taga T, Ito E, Horibe K, Miyano S, Adachi S, Taki T, Ogawa S, Hayashi Y.
  • Journal Title: Genes Chromosomes Cancer Volume: 62 Issue: 7 Pages: 412-422
  • DOI: 10.1002/gcc.23147
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Genome-wide DNA Methylation Analysis in Pediatric Acute Myeloid Leukemia. (2022)
  • Author(s): Yamato G, Kawai T, Shiba N, Ikeda J, Hara Y, Ohki K, Tsujimoto SI, Kaburagi T, Yoshida K, Shiraishi Y, Miyano S, Kiyokawa N, Tomizawa D, Shimada A, Sotomatsu M, Arakawa H, Adachi S, Taga T, Horibe K, Ogawa S, Hata K, Hayashi Y.
  • Journal Title: Blood advances Volume: in press Issue: 11 Pages: 3207-3219
  • DOI: 10.1182/bloodadvances.2021005381
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] YM155 and chrysin cooperatively suppress survivin expression in SMARCB1/INI1-deficient tumor cells (2022)
  • Author(s): Yoshino Yuki、Goto Hiroaki、Ito Mieko、Tsurusaki Yoshinori、Takita Junko、Hayashi Yasuhide、Yanagimachi Masakatsu
  • Journal Title: Medical Oncology Volume: 39 Issue: 12 Pages: 234-234
  • DOI: 10.1007/s12032-022-01843-4
  • Peer Reviewed
• [Journal Article] Clinical significance of RAS pathway alterations in pediatric acute myeloid leukemia. (2021)
  • Author(s): Kaburagi T, Yamato G, Shiba N, Yoshida K, Hara Y, Tabuchi K, Shiraishi Y,Ohki K, Sotomatsu M, Arakawa H, Matsuo H, Shimada A, Taki T, Kiyokawa N, Tomizawa D, Horibe K, Miyano S, Taga T, Adachi S, Ogawa S, Hayashi Y.
  • Journal Title: Haematologica Volume: － Issue: 3 Pages: 583-592
  • DOI: 10.3324/haematol.2020.269431
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] C/EBPβ induces B‐cell acute lymphoblastic leukemia and cooperates with <i>BLNK</i> mutations (2021)
  • Author(s): Kurata Morito、Onishi Iichiro、Takahara Tomoko、Yamazaki Yukari、Ishibashi Sachiko、Goitsuka Ryo、Kitamura Daisuke、Takita Junko、Hayashi Yasuhide、Largaesapda David A、Kitagawa Masanobu、Nakamura Takuro
  • Journal Title: Cancer Science Volume: 112 Issue: 12 Pages: 4920-4930
  • DOI: 10.1111/cas.15164
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Characteristics of genetic alterations of peripheral T‐cell lymphoma in childhood including identification of novel fusion genes: the Japan Children’s Cancer Group (JCCG) (2021)
  • Author(s): Ohki K、Kiyokawa N、Watanabe S、Iwafuchi H、Nakazawa A、Ishiwata K、Ogata‐Kawata Hiroko、Nakabayashi K、Okamura K、Tanaka F、Fukano R、Hata K、Mori T、Moriya Saito Akiko、Hayashi Y、Taga T、Sekimizu M、Kobayashi R、for the Japan Children’s Cancer Study Group (JCCG)
  • Journal Title: British Journal of Haematology Volume: 194 Issue: 4 Pages: 718-729
  • DOI: 10.1111/bjh.17639
  • Peer Reviewed
• [Journal Article] Predictive factors for the development of leukemia in patients with transient abnormal myelopoiesis and Down syndrome (2021)
  • Author(s): Yamato G, Deguchi T, Terui K, Toki T, Watanabe T, Imaizumi T, Hama A, Iwamoto S, Hasegawa D, Ueda T, Yokosuka T, Tanaka S, Yanagisawa R, Koh K, Saito AM, Horibe K, Hayashi Y, Adachi S, Mizutani S, Taga T, Ito E, Watanabe K, Muramatsu H.
  • Journal Title: Leukemia. Volume: 35 Issue: 5 Pages: 1480-1484
  • DOI: 10.1038/s41375-021-01171-y
  • Peer Reviewed
• [Journal Article] Association of high-risk neuroblastoma classification based on expression profiles with differentiation and metabolism (2021)
  • Author(s): Kimura Shunsuke、Sekiguchi Masahiro、Watanabe Kentaro、Hiwatarai Mitsuteru、Seki Masafumi、Yoshida Kenichi、Isobe Tomoya、Shiozawa Yusuke、Suzuki Hiromichi、Hoshino Noriko、Hayashi Yasuhide、Oka Akira、Miyano Satoru、Ogawa Seishi、Takita Junko
  • Journal Title: PLOS ONE Volume: 16 Issue: 1 Pages: 0245526-0245526
  • DOI: 10.1371/journal.pone.0245526
  • Peer Reviewed / Open Access
• [Journal Article] Clinical features of 35 Down syndrome patients with transient abnormal myelopoiesis at a single institution. (2021)
  • Author(s): Yamato G, Park MJ, Hayashi Y, et al.
  • Journal Title: Int J Hematol. Volume: 113 Issue: 5 Pages: 662-667
  • DOI: 10.1007/s12185-020-03066-7
  • Peer Reviewed
• [Journal Article] Droplet digital polymerase chain reaction assay for the detection of the minor clone of KIT D816V in paediatric acute myeloid leukaemia especially showing RUNX1-RUNX1T1 transcripts. (2021)
  • Author(s): Sasaki K, Tsujimoto S, Miyake M, Uchiyama Y, Ikeda J, Yoshitomi M, Shimosato Y, Tokumasu M, Matsuo H, Yoshida K, Ohki K, Kaburagi T, Yamato G, Hara Y, Takeuchi M, Kinoshita A, Tomizawa D, Taga T, Adachi S, Tawa A, Horibe K, Hayashi Y, Matsumoto N, Ito S, Shiba N.
  • Journal Title: British Journal of Haematology Volume: 194(2) Issue: 2 Pages: 414-422
  • DOI: 10.1111/bjh.17569
  • Peer Reviewed
• [Journal Article] Impact of immunophenotypic characteristics on genetic subgrouping in childhood acute lymphoblastic leukemia: Tokyo Children’s Cancer Study Group ( TCCSG ) study L04 ‐16 (2020)
  • Author(s): Ohki Kentaro, Takahashi Hiroyuki, Fukushima Takashi, Kiyokawa Nobutaka. et al.
  • Journal Title: Genes, Chromosomes and Cancer Volume: － Issue: 10 Pages: 551-561
  • DOI: 10.1002/gcc.22858
  • Peer Reviewed
• [Journal Article] 白血病とリンパ腫の染色体・FISH分析と臨床への応用 (2020)
  • Author(s): 23.林 泰秀.
  • Journal Title: 日本臨牀 Volume: 78 Pages: 183-192
• [Journal Article] DNA methylation-based classification reveals difference between pediatric T-cell acute lymphoblastic leukemia and normal thymocytes (2019)
  • Author(s): Kimura Shunsuke、Ogawa Seishi., et al.
  • Journal Title: Leukemia Volume: 34 Issue: 4 Pages: 1163-1168
  • DOI: 10.1038/s41375-019-0626-2
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] UBTF-ITDは小児急性骨髄性白血病の予後因子である (2022)
  • Author(s): 鏑木多映子, 大和玄季, 柴徳生, 吉田健一, 石北悦子, 原勇介, 白石友一, 大木健太郎, 河崎裕英, 外松学, 滝沢琢己, 滝智彦, 清河信 敬, 富澤大輔, 堀部敬三, 宮野悟, 多賀崇, 足立壮一, 小川誠司, 林泰秀
  • Organizer: 第84回日本血液学会学術集会
• [Presentation] DNAメチル化パターンによるKMT2A-MLLT3陽性小児急性骨髄性白血病患者の予後予測 (2022)
  • Author(s): 大和玄季, 河合智子, 柴 徳生, 原勇介, 大木健太郎, 鏑木多映子, 石北悦子, 吉田健一, 白石友一, 宮野悟, 清河信敬, 富澤 大輔 , 嶋田明, 外松学, 滝沢琢己 , 足立壮一, 多賀崇 , 堀部敬三, 小川誠司, 秦健一郎, 林泰秀
  • Organizer: 第84回日本血液学会学術集会
• [Presentation] 1.Prognosis of pediatric acute myeloid leukemia with KMT2A-MLLT3 according to DNA methylation patterns: JCCG JPLSG AML-05 study. (2022)
  • Author(s): Yamato G, Kawai T, Shiba N, Hara Y, Ohki K, Kaburagi T, Ishikita E, Kawasaki H, Yoshida K, Shiraishi Y, Miyano S, Kiyokawa N, Tomizawa D, Shimada A, Sotomatsu M, Takizawa T, Adachi S, Taga T, Horibe K, Ogawa S, Hata K, Hayashi Y.
  • Organizer: 64th American Society of Hematology Annual Meeting. 米国
  • Int'l Joint Research
• [Presentation] 小児急性骨髄性白血病における全ゲノムDNAメチル化解析 -The JCCG-JPLSG AML-05 study- (2021)
  • Author(s): 1.大和玄季、河合智子, 柴 徳生, 原勇介, 大木健太郎, 鏑木多映子, 吉田健一, 白石友一, 宮野悟, ,小川誠司, 秦健一郎, 林泰秀。
  • Organizer: 第83回日本血液学会
• [Presentation] 一過性骨髄異常増殖症患者151例におけるサイトカイン解析 (2021)
  • Author(s): 2.大和玄季、朴明子、嶋田明、柴徳生、山田佳之、渡邉智之、照井君典、村松秀城、渡邉健一郎、林泰秀
  • Organizer: 第６3回日本小児血液・がん学会
• [Presentation] Cytokine analysis in 154 patients with transient abnormal myelopoiesis: JCCG JPLSG TAM-10 clinical study (2021)
  • Author(s): 3.Yamato G, Park MJ, Shimada A, Shiba N, Yamada Y, Terui K, Ito E, Muramatsu H, Watanabe K, Hayashi Y
  • Organizer: 63rd American Society of Hematology Annual Meeting
• [Presentation] 小児急性骨髄性白血病においてDOCK1遺伝子高発現は予後不良因子となり得る (2020)
  • Author(s): 吉富誠弘、富澤大輔、足立壮一、林 泰秀、多賀崇、柴徳生。
  • Organizer: 第６2回日本小児血液・がん学会
• [Presentation] 小児急性前骨髄性白血病に対するカスタムパネルを用いた網羅的遺伝子解析 -the JCCG study, JPLSG AML-P05-。 (2020)
  • Author(s): 大和玄季, 上野博雄、高橋博之、柴徳生、富澤大輔、足立壮一、多賀崇、林泰秀。 2020年11月20~22日、郡山、国内、WEB口演。
  • Organizer: 第６1回日本小児血液・がん学会