Analysis of genome structure of relapse/refractory acute leukemia by next generation sequencing

• Project/Area Number: 16K09836
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Hematology
• Research Institution: Hokkaido University
• Principal Investigator: Onozawa Masahiro 北海道大学, 大学病院, 助教 (70455632)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2018: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2017: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2016: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
• Keywords: ゲノム損傷 / 再発難治性白血病 / DNA二重鎖切断 / ゲノム修復 / 白血病 / ゲノム / 遺伝子 / 癌 / 内科

Outline of Final Research Achievements

This project aim to analyze the change of genome structure of acute leukemia due to DNA damage induced by irradiation or chemotherapy using next generation sequencing. Accumulation of somatic mutations due to DNA repair might contribute leukemogenesis or acquired resistance to chemotherapy through loss of expression of tumor suppressor. We confirmed that artificially induced site specific DNA double strand break were repaired by various length of deletion or templated sequence insertion. We analyze 2 sets of human leukemic cell lines and its daughter clone by whole genome sequencing. We examined several methodology to elute meaningful sequence reads which contained tumor specific somatic indels.

Academic Significance and Societal Importance of the Research Achievements

腫瘍細胞ではゲノム損傷修復機能が活性化しており、放射線や化学療法で治療しても、DNAのダメージを乗り越えて生き残る細胞が再発をきたすと考えられる。一塩基置換による遺伝子機能の活性化とは別に、放射線や化学療法によるDNA切断部位は欠失挿入による傷をゲノム上に残して修復される。正常細胞と比べて、腫瘍細胞のゲノム構造がどれほど傷ついた状態であるかを数値化することができれば、その値は治療反応性や予後と相関する可能性がある。このコンセプトを確かめるためには、現在の次世代シークエンスよりも、より長い塩基数をより正確に安価に読み取る技術と専用の解析アルゴリズムの開発が必要であることも明らかとなった。

Research Products

• [Journal Article] Development of a Fluorescence in Situ Hybridization Probe for Detecting IKZF1 Deletion Mutations in Patients with Acute Lymphoblastic Leukemia (2018)
  • Author(s): Hashiguchi Junichi、Onozawa Masahiro、Oguri Satoshi、Fujisawa Shinichi、Tsuji Masahisa、Okada Kohei、Nakagawa Masao、Hashimoto Daigo、Kahata Kaoru、Kondo Takeshi、Shimizu Chikara、Teshima Takanori
  • Journal Title: The Journal of Molecular Diagnostics Volume: 20 Issue: 4 Pages: 446-454
  • DOI: 10.1016/j.jmoldx.2018.02.005
  • NAID: 120006649182
  • Peer Reviewed
• [Journal Article] Wilms Tumor 1 Expression at Diagnosis Correlates With Genetic Abnormalities and Polymorphism But Is Not Independently Prognostic in Acute Myelogenous Leukemia: A Hokkaido Leukemia Net Study. (2018)
  • Author(s): Hidaka D, Onozawa , et al.
  • Journal Title: Clin Lymphoma Myeloma Leuk. Volume: 18 Issue: 11 Pages: e469-e479
  • DOI: 10.1016/j.clml.2018.07.291
  • Peer Reviewed
• [Journal Article] Quantitative detection of IKZF1 deletion by digital PCR in patients with acute lymphoblastic leukemia (2018)
  • Author(s): Hashiguchi J, Onozawa M, Okada K, Amano T, Hatanaka KC, Nishihara H, Sato N, Teshima T
  • Journal Title: Int J Lab Hematol Volume: 41 Issue: 2
  • DOI: 10.1111/ijlh.12945
  • Peer Reviewed
• [Journal Article] Templated Sequence Insertion Polymorphisms in the Human Genome. (2016)
  • Author(s): Masahiro Onozawa
  • Journal Title: Frontiers in Chemistry Volume: 4 Pages: 43-43
  • DOI: 10.3389/fchem.2016.00043
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Presentation] WT1 expression at onset of AML correlated collaborative cytogenetic abnormality and WT1 SNP status (2018)
  • Author(s): Masahiro Onozawa, et al.
  • Organizer: 第80回日本血液学会学術集会