Analysis of residual disease using newly developed single cell RNA sequencing.

• Project/Area Number: 17K09926
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Hematology
• Research Institution: Kyoto University
• Principal Investigator: Nakagawa Masahiro 京都大学, 医学研究科, 特定講師 (10431850)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2019: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2017: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 単一細胞解析 / 白血病 / 骨髄異形成症候群 / シングルセル解析 / 単一細胞 / 遺伝子発現

Outline of Final Research Achievements

We have developed a new single-cell analysis platform that can identify mutations with high sensitivity for the analysis of refractory clones in intratumor heterogeneity, which is a major factor in the therapy resistance of hematological malignancies. Using this method, hematopoietic progenitors derived from the bone marrow of healthy individuals or patients with hematological malignancies were analyzed. We found that apparently normal hematopoietic progenitors show aberrant expression profiles compared to normal cells in healthy controls without driver mutations. These results suggest that a favorable environment for the clone is important for their expansion and was considered to be an important finding for the realization of a new therapeutic intervention.

Academic Significance and Societal Importance of the Research Achievements

本研究で開発した、高感度にドライバー変異を同定可能な単一細胞RNAシーケンス技術の適用範囲は極めて広い。変異細胞を区別して解析可能なため、腫瘍の初期発生から、多様な変異を有する細胞が混在する腫瘍内多様性の解析などで、変異の及ぼす細胞自律性の影響だけで無く、腫瘍細胞と環境の相互作用などの非細胞自律性の解析も可能となる。腫瘍のクローン進化のさらなる理解と新規の治療法の開発に向けた大きな意義ある一歩であると考えられる。

Research Products

• [Presentation] Analysis of mechanisms underlying clonal evolution of AML by a new single-cell sequencing platform (2019)
  • Author(s): Ryosaku Inagaki, Masahiro M Nakagawa, Yasuhito Nannya, Xingxing Qi, Zhao Lanying, June Takeda, Akinori Yoda, Ayana Kon, Hisashi Tsurumi, Hideki Makishima, and Seishi Ogawa
  • Organizer: 61th ASH Annual Meeting
  • Int'l Joint Research
• [Presentation] Analysis of mechanisms underlying clonal evolution in MDS to sAML transformation by single-cell sequencing (2019)
  • Author(s): Ryosaku Inagaki, Masahiro M Nakagawa, Yasuhito Nannya, Xingxing Qi, Zhao Lanying, June Takeda, Akinori Yoda, Ayana Kon, Hideki Makishima, and Seishi Ogawa
  • Organizer: 第78回日本癌学会学術総会
• [Presentation] Analysis of mechanisms underlying clonal evolution of AML by a new single-cell sequencing platform (2019)
  • Author(s): Ryosaku Inagaki, Masahiro M Nakagawa, Yasuhito Nannya, Xingxing Qi, Zhao Lanying, June Takeda, Akinori Yoda, Ayana Kon, Hisashi Tsurumi, Hideki Makishima, and Seishi Ogawa
  • Organizer: 第81回日本血液学会学術総会
• [Presentation] Intratumor heterogeneity and clonal evolution of myeloid malignancies revealed by single-cell RNA/DNA sequencing. (2019)
  • Author(s): Masahiro M Nakagawa, Ryosaku Inagaki, Yasuhito Nannya,, Zhao Lanying, June Takeda1, Xingxing Qi, Akinori Yoda, Ayana Kon, Hisashi Tsurumi, Hideki Makishima, Shuichi Matsuda and Seishi Ogawa
  • Organizer: 第78回日本癌学会学術総会
• [Presentation] Pathogenesis of clonal hematopoiesis revealed by single-cell RNA/DNA sequencing. (2019)
  • Author(s): Masahiro M Nakagawa, Ryosaku Inagaki, Yasuhito Nannya, Yutaka Kuroda, Zhao Lanying, June Takeda1, Xingxing Qi, Akinori Yoda, Ayana Kon, Hisashi Tsurumi, Hideki Makishima, Shuichi Matsuda and Seishi Ogawa
  • Organizer: 第81回日本血液学会学術総会
• [Presentation] Mechanisms of the clonal evolution of MDS as revealed by single-cell sequencing. (2018)
  • Author(s): Masahiro M Nakagawa, Ryosaku Inagaki, Yasuhito Nannya, Zhao Lanying, June Takeda, Akinori Yoda, Ayana Kon, Tetsuichi Yoshizato, Hideki Makishima, and Seishi Ogawa
  • Organizer: 第77回日本癌学会学術総会
• [Presentation] Mechanisms of the clonal evolution of MDS as revealed by single-cell sequencing. (2018)
  • Author(s): Masahiro M Nakagawa, Ryosaku Inagaki, Yasuhito Nannya, Zhao Lanying, June Takeda, Akinori Yoda, Ayana Kon, Tetsuichi Yoshizato, Hisashi Tsurumi, Hideki Makishima, and Seishi Ogawa
  • Organizer: 第80回日本血液学会学術総会
• [Presentation] Analysis of clonal evolution/heterogeneity of MDS by simultaneous detection of both mutation and gene expression by single cell sequencing (2018)
  • Author(s): Masahiro Marshall Nakagawa, Ryosaku Inagaki, Yasuhito Nannya, Lanying Zhao, Yutaka Kuroda, June Takeda, Xingxing Qi, Akinori Yoda, Ayana Kon, Hisashi Tsurumi, Hideki Makishima, Shuichi Matsuda and Seishi Ogawa
  • Organizer: 60th ASH Annual Meeting
  • Int'l Joint Research
• [Presentation] Mechanisms of the clonal evolution of myeloid malignancies as revealed by single-cell sequencing. (2017)
  • Author(s): Masahiro M Nakagawa, Ryosaku Inagaki, Yasuhito Nannya, Keisuke Kataoka, Tetsuichi Yoshizato, Hideki Makishima, Kenichi Yoshida, and Seishi Ogawa
  • Organizer: 日本癌学会
• [Presentation] Mechanisms of the clonal evolution of MDS as revealed by single-cell sequencing. (2017)
  • Author(s): Masahiro M Nakagawa, Ryosaku Inagaki, Yasuhito Nannya, Keisuke Kataoka, Tetsuichi Yoshizato, Hideki Makishima, Kenichi Yoshida, and Seishi Ogawa
  • Organizer: 日本血液学会