Development of GBM-therapeutic recombinant viruses that encodes miR-dependent genome-editing system.

• Project/Area Number: 20H03559
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 51030:Pathophysiologic neuroscience-related
• Research Institution: Hokkaido University
• Principal Investigator: Kondo Toru 北海道大学, 遺伝子病制御研究所, 教授 (30270573)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥13,650,000 (Direct Cost: ¥10,500,000、Indirect Cost: ¥3,150,000); Fiscal Year 2022: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2021: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2020: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
• Keywords: 脳腫瘍 / ゲノム編集 / マイクロRNA / 組換えウイルス / 癌幹細胞 / Glioblastoma (GBM) / miRNA / AAV / GBM initiating cell / 担癌マウス / miR

Outline of Research at the Start

研究代表者らが開発を進めているmiR依存的にGIC機能因子をゲノム編集するベクターシステムを組み込んだ複数の新規組換えウイルスを作製し、様々な正常細胞に影響がなくGICを特異的に傷害することを確認する。次に、これら組換えウイルスを担がんマウスに感染させ、それらの個体への影響と抗腫瘍効果を検討し、臨床試験へ適用可能な安全性と治療効果の高い新規グリオーマ治療用ウイルスの作製を目的として本研究を遂行する。

Outline of Final Research Achievements

We successfully generated novel adeno-associated virus (AAV) that encodes the expression cassettes of both Cas9 with complementary sequence of microRNA (miR), which is expressed in non-tumor cells including nerural stem cells (NSC) but not in glioblastoma (GBM) stem cells (GICs), and guide RNA for a GIC functional factor. We found that this AAV killed GICs but not NSCs in culture. We confirmed that the intravenously injected AAV prevented GIC tumorigenesis in GIC-transplanted mice and extended their survival. Thus, we succeeded to generate the novel genome-edited AAV that specifically kills GBM in a miR expression-dependent manner.

Academic Significance and Societal Importance of the Research Achievements

本研究は、細胞（種）やその状況に発現変化するmiRを用いて任意の標的細胞で外来遺伝子の発現制御が可能であることを証明した。同様の方法を用いることによりmiRにより制御を受ける全ての細胞を対象とした様々な研究への応用が可能である。更に、今回作製したmiRの発現逆依存的にGIC機能因子をゲノム編集するAAVは、GBMと適用可能な他の悪性腫瘍に対する新規遺伝子治療用ウイルスとなる可能性が高く、その社会的意義は非常に大きい。

Research Products

• [Journal Article] Selective vulnerability of human-induced pluripotent stem cells to dihydroorotate dehydrogenase inhibition during mesenchymal stem/stromal cell purification (2023)
  • Author(s): Al-Akashi Ziadoon、Zujur Denise、Kamiya Daisuke、Kato Tomohisa、Kondo Toru、Ikeya Makoto
  • Journal Title: Frontiers in Cell and Developmental Biology Volume: 11 Pages: 1089945-1089945
  • DOI: 10.3389/fcell.2023.1089945
  • Peer Reviewed / Open Access
• [Journal Article] Glioblastoma-initiating cell heterogeneity generated by the cell-of-origin, genetic/epigenetic mutation and microenvironment (2021)
  • Author(s): Kondo Toru
  • Journal Title: Seminars in Cancer Biology Volume: in press Pages: 176-183
  • DOI: 10.1016/j.semcancer.2020.12.003
  • Peer Reviewed
• [Presentation] Novel genome-editing AAVs that selectively eradicate glioblastoma-initiating cells (2022)
  • Author(s): Toru Kondo
  • Organizer: 6th European Congress on Neurology and Brain Disorders
  • Int'l Joint Research / Invited
• [Presentation] Novel genome-editing AAVs that selectively eradicate glioblastoma-initiating cells (2022)
  • Author(s): Toru Kondo
  • Organizer: The 81st Annual Meeting of the Japanese Cancer Association