Preparation of and drug discovery studies by hepatobiliary pancreatic cancer stem cell models derived from organoid and induced pluripotent stem cells

• Project/Area Number: 19K06457
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 42040:Laboratory animal science-related
• Research Institution: Okayama University
• Principal Investigator: Iwasaki Yoshiaki 岡山大学, 保健管理センター, 教授 (00314667)
• Co-Investigator(Kenkyū-buntansha): 笠井 智成 岡山大学, 中性子医療研究センター, 准教授 (30530191)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2021: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 肝胆膵がん / がん幹細胞 / オルガノイド / 人工多能性幹細胞 / 多能性幹細胞 / 動物モデル

Outline of Research at the Start

これまでわれわれが確立してきた、iPS細胞からがんの元や、がんの転移・再発の原因となる細胞（がん幹細胞）のモデルを作製する手法を用いて、肝がんのがん幹細胞モデルを作製してきた。しかしながら、iPS細胞から肝細胞への分化の効率、がん幹細胞モデルの安定的維持、動物モデル作製の際のがん幹細胞の移植効率に問題があった。そこで本研究では、オルガノイド（試験管内でつくられる立体的なミニ臓器）作製技術の応用により、これら問題の解決を図るとともに、がん幹細胞モデル作製技術と組み合わせて、肝臓・胆のう・胆管・膵臓のがん幹細胞動物モデルを作製し、新しい薬剤の開発に応用する。

Outline of Final Research Achievements

We induced differentiation of mouse induced pluripotent stem cells into liver and bile duct progenitors and pancreatic progenitors via embryonic endoderm, and induced tumorigenesis of cells at each stage using a method to generate cancer stem cell models. Organoids at each stage of differentiation were also generated and induced to become cancerous in the same manner. These cells and organoids were transplanted subcutaneously and into the liver of mice to form masses. The efficiency of organoid and tumor formation was higher in undifferentiated cells, but the efficiency of tumor formation was still low, requiring further investigation of conditions for induction of differentiation and tumorigenesis. On the other hand, organoids of mouse liver and bile duct progenitor cells were induced to become cancerous and transplanted into mouse liver, but the conditions at each step need to be improved for stable tumor formation.

Academic Significance and Societal Importance of the Research Achievements

遺伝的に明確かつ均一ながん幹細胞モデルの作成は、がんの生物学的な解析などの基礎的な研究に重要であり、その解析成果による臨床応用も期待される。さらに、がん幹細胞動物モデルの作成は、診断及び創薬などの臨床応用に直結する可能性を秘めている。本研究ではこれらがん幹細胞およびそのオルガノイドモデルの作成からがん幹細胞動物モデルの作成を試みたものであり、その過程に影響を及ぼす種々の要因が明らかになった。これらの要因の更なる究明によるより効率的ながん幹細胞モデルの作成が今後期待される。

Research Products

• [Journal Article] Temporal and Locational Values of Images Affecting the Deep Learning of Cancer Stem Cell Morphology (2022)
  • Author(s): Hanai Yumi, Ishihata Hiroaki, Zhang Zaijun, Maruyama Ryuto, Kasai Tomonari, Kameda Hiroyuki, Sugiyama Tomoyasu
  • Journal Title: Biomedicines Volume: 10 Issue: 5 Pages: 941-941
  • DOI: 10.3390/biomedicines10050941
  • Peer Reviewed / Open Access
• [Journal Article] A novel model of liver cancer stem cells developed from induced pluripotent stem cells (2020)
  • Author(s): Afify SM, Sanchez Calle A, Hassan G, Kumon K, Nawara HM, Zahra MH, Mansour HM, Khayrani AC, Alam MJ, Du J, Seno A, Iwasaki Y, Seno M
  • Journal Title: Br J Cancer Volume: 122 Issue: 9 Pages: 1378-1390
  • DOI: 10.1038/s41416-020-0792-z
  • NAID: 120006949462
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Metastasis of Cancer Stem Cells Developed in the Microenvironment of Hepatocellular Carcinoma (2019)
  • Author(s): Afify SM, Hassan G, Osman A, Calle AS, Nawara HM, Zahra MH, El-Ghlban S, Mansour H, Alam MJ, Abu Quora HA, Du J, Seno A, Iwasaki Y, Seno M.
  • Journal Title: Bioengineering Volume: 6 Issue: 3 Pages: 73-73
  • DOI: 10.3390/bioengineering6030073
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Method to Convert Stem Cells Into Cancer Stem Cells (2019)
  • Author(s): Afify SM, Chen L, Yan T, Calle AS, Nair N, Murakami C, Zahra MH, Okada N, Iwasaki Y, Seno A, Seno M.
  • Journal Title: Methods Protoc Volume: 2 Issue: 3 Pages: 71-71
  • DOI: 10.3390/mps2030071
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Cancer Stem Cell Induction From Mouse Embryonic Stem Cells (2019)
  • Author(s): Seno A, Murakami C, El-Aarag B, Iwasaki Y, Ohara T, Seno M.
  • Journal Title: Oncol Lett Volume: 18 Pages: 2756-2762
  • DOI: 10.3892/ol.2019.10614
  • NAID: 120006823176
  • Peer Reviewed / Int'l Joint Research