How does aberrant B lymphocyte produce a origin of multiple myeloma cells?

• Project/Area Number: 20K08738
• 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: Fukushima Medical University
• Principal Investigator: Sakai Akira 福島県立医科大学, 医学部, 教授 (70284221)
• Co-Investigator(Kenkyū-buntansha): 阿部 悠 福島県立医科大学, 医学部, 助教 (00722472) ; 津山 尚宏 福島県立医科大学, 医学部, 准教授 (10335747)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2022: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2021: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 成熟Bリンパ球 / 多発性骨髄腫 / iPS細胞 / 再プログラミング / 形質転換 / 骨髄先駆細胞 / 骨髄前駆細胞 / 染色体転座 / 造血前駆細胞

Outline of Research at the Start

我々は成熟Bリンパ球のがん化の機序を解明するため正常Bリンパ球からiPS細胞 (BiPSCs) を樹立した。これらBiPSCsはDNA２本鎖切断に関与するAIDを発現誘導でき、さらにゲノム編集により染色体転座t(11;14)を有しp53遺伝子を欠失するがストローマ細胞との共培養で造血前駆細胞に分化する。本研究では、多発性骨髄腫 (MM) の腫瘍起源となる異常Bリンパ球は、成熟Bリンパ球が再プログラミングによってポテンシャルを獲得し染色体異常を生じた細胞であることを、BiPSCsのin vitroでのBリンパ球への分化能および免疫不全マウスへの移植によって検証を行う。

Outline of Final Research Achievements

The onset of MM is often caused by a reciprocal chromosomal translocation (cTr) between chr 14 with IgH and chr 11 with CCND1. We propose that mature B cells gain potential to transform by reprograming, and then chromosomal aberrations cause the development of abnormal B cells as a myeloma-initiating cell during B cell redifferentiation. To study myeloma-initiating cells, we have already established normal B cell-derived induced pluripotent stem cells (BiPSCs). Here we established two BiPSCs with reciprocal cTr t(11;14) using the CRISPR/Cas9 system. Furthermore, p53 was deleted using the CRISPR/Cas9 system in BiPSC13 with t(11;14). These BiPSCs differentiated into hematopoietic progenitor cells (HPCs). However, unlike cord blood, those HPCs did not differentiated into B lymphocytes by co-culture with BM stromal cell. Therefore, further ingenuity is required to differentiate those BiPSCs-derived HPCs into B lymphocytes.

Academic Significance and Societal Importance of the Research Achievements

本研究では、多発性骨髄腫 (MM) の腫瘍起源となる異常Bリンパ球は、成熟Bリンパ球が再プログラミングによってポテンシャルを獲得し染色体異常を生じた細胞であることの検証を行う。確認できれば、生体内で加齢や慢性炎症による正常細胞のエピジェネティックな変化が細胞のがん化に繋がる機序の１つであることを支持する腫瘍化のモデルであり、がん発生の予防方法の開発に繋がる。

Research Products

• [Journal Article] Chromosomal translocation t(11;14) and p53 deletion induced by the CRISPR/Cas9 system in normal B cell-derived iPS cells (2021)
  • Author(s): Yusuke Azami, Naohiro Tsuyama, Yu Abe, Misaki Sugai-Takahashi, Ken-Ichi Kudo, Akinobu Ota, Karnan Sivasundaram , Moe Muramatsu, Tomonari Shigemura, Megumi Sasatani, Yuko Hashimoto, Shigehira Saji, Kenji Kamiya, Ichiro Hanamura, Takayuki Ikezoe, Masafumi Onodera, Akira Sakai.
  • Journal Title: Sci Rep Volume: 11 Issue: 1 Pages: 5216-5216
  • DOI: 10.1038/s41598-021-84628-5
  • Peer Reviewed / Open Access
• [Presentation] Attempts to induce t(11;14) during the redifferentiation of iPS cells into B cells. (2022)
  • Author(s): Misaki Takahashi, Naohiro Tsuyama, Ken-ichi Kudo, Yusuke Azami, Miwa Fukami, Takayuki Ikezoe, Masafumi Onodera, Akira Sakai.
  • Organizer: 第84回日本血液学会学術集会
• [Presentation] Analysis of differential gene expression in t(11;14) carrying iPS cells induced by genome editing. (2022)
  • Author(s): Naohiro TSUYAMA, Kenichi KUDO, Misaki SUGAI-TAKAHASHI, Yusuke AZAMI, Kenji KAMIYA, Akira SAKAI.
  • Organizer: 第65回日本放射線影響学会
• [Presentation] Study for exploring myeloma-initiating cell using normal B cell-derived iPS cells (2021)
  • Author(s): Yusuke Azami, Naohiro Tsuyama, Yu Abe, Misaki Sugai-Takahashi, Ken-ichi Kudo, Miwa Fukami, Akinobu Ota, Karnan Sivasundaram, Moe Muramatsu, Tomonari Shigemura, Megumi Sasatani, Yuko Hashimoto, Shigehira Saji, Kenji Kamiya, Ichiro Hanamura, Takayuki Ikezoe, Masafumi Onodera, Akira Sakai
  • Organizer: 第83回日本血液学会学術集会
• [Presentation] 正常Bリンパ球由来iPS細胞を用いた骨髄腫起源細胞の探索 (2021)
  • Author(s): 坂井 晃
  • Organizer: 第46回日本骨髄腫学会学術集会 会長講演
  • Invited
• [Presentation] Chromosomal translocation t(11;14) induced by Cre-loxP system in normal B cell-derived iPS cells (2020)
  • Author(s): Misaki Sugai, Naohiro Tsuyama, Yu Abe, Yusuke Azami, Akinobu Ota, Karnan Sivasundaram, Moe Muramatsu, Tomonari Shigemura, Megumi Sasatani, Yuko Hashimoto, Kenji Kamiya, Ichiro Hanamura, Takayuki Ikezoe, Masafumi Onodera, Akira Sakai.
  • Organizer: 第82回日本血液学会学術集会
• [Presentation] Chromosomal Translocation t(11;14) Induced by the Cre-loxP System in Normal B cell-derived iPS Cells for the Study of Myeloma-initiating Cells (2020)
  • Author(s): Misaki Sugai, Naohiro Tsuyama, Yu Abe, Yusuke Azami, Kenichi Kudo, Akinobu Ota, Karnan Sivasundaram, Moe Muramatsu, Tomonari Shigemura, Megumi Sasatani, Yuko Hashimoto, Kenji Kamiya, Ichiro Hanamura, Takayuki Ikezoe, Masafumi Onodera, Akira Sakai.
  • Organizer: 62th ASH Annual Meeting