Tumor-tropic liposome-mediated therapeutic delivery of mRNA for the treatment of adolescent and young adult cancers.

• Project/Area Number: 20K08227
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 52050:Embryonic medicine and pediatrics-related
• Research Institution: Shinshu University
• Principal Investigator: Saito Shoji 信州大学, 学術研究院医学系(医学部附属病院), 講師 (10623762)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2022: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2021: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2020: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: リポソーム / ｍRNA医薬 / Caspase 9 / アポトーシス / 遺伝子治療 / ナノ粒子 / mRNA医薬 / 自殺遺伝子 / 乳がん / AYA世代がん / 小児がん / 新規がん治療法

Outline of Research at the Start

近年、ナノ粒子を利用した薬剤デリバリーシステム（DDS）が注目されている。標的病変に優先的に集積する傾向を生かして、安全かつ有効に薬剤の効果を引き出すことが可能となってきた。われわれは、腫瘍細胞の細胞膜の特異性に注目することで、腫瘍細胞に優先的にとりこまれ集積されるナノ粒子を開発した。本研究では、このナノ粒子に治療遺伝子を搭載し、腫瘍細胞内で治療遺伝子を発現させることで、腫瘍細胞の遺伝子を改変し特異的な抗腫瘍効果をもたらす、革新的がん治療法の開発を目的としている。最終的には、この治療技術を用いて、難治性小児・思春期・若年成人がんの治療成績向上に貢献することを目的とする。

Outline of Final Research Achievements

Cancer gene therapy can be potential therapeutics for advanced/refractory cancers. Combining tumor-tropic lipid nanoparticles (LNPs) and inducible caspase-9 (iC9) mRNA, we aimed to develop a novel treatment strategy for refractory breast cancer. LNP’s anti-tumor effects were tested in vitro in three breast cancer cell lines: MDA-MB231, SKBR3, and MCF-7. LNPs could efficiently deliver encapsulated GFP mRNA to all three cancer cell lines. Furthermore, LNPs encapsulated with iC9 mRNA (iC9-LNPs) and CID showed cytotoxic activity against all cancer cell lines in vitro. Interestingly, susceptibility to iC9 gene therapy was heterogeneous among cancer cell lines. Quantification of apoptosis-related genes suggested that a high BAX/Bcl-2 ratio might be associated with iC9-LNP+CID susceptibility. Thus, cancer gene therapy using iC9-LNPs and CID could be a promising alternative for the treatment of breast cancers, especially for aggressive breast cancers.

Academic Significance and Societal Importance of the Research Achievements

本研究において、難治性乳がんに対する、腫瘍指向性脂質ナノ粒子(LNP)と誘導性カスパーゼ9(iC9)mRNAを組み合わせた新規遺伝子治療法の開発に成功した。本研究では乳がんを対象に治療効果を検証したが、本遺伝子治療法は、理論的にはがん種によらず有効性が示せるため、さまざまながん種に対する治療法に応用できる。また、アポトーシス関連の解析を行ったところ、乳がんにおいてはより悪性度の高いがん種において感受性が高いことを示した。このため、本疾患は難治性がんに対する有望な治療法として応用できる可能性がある。

Research Products

• [Journal Article] Non-viral inducible caspase 9 mRNA delivery using lipid nanoparticles against breast cancer: An in-vitro study (2022)
  • Author(s): Nakashima Ikumi、Saito Shoji、Akahoshi Eiichi、Yagyu Shigeki、Sugano-Ishihara Mitsuko、Nakazawa Yozo
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 635 Pages: 144-153
  • DOI: 10.1016/j.bbrc.2022.09.105
  • Peer Reviewed / Open Access
• [Journal Article] AMLに対する抗体・免疫細胞療法の動向 (2022)
  • Author(s): 齋藤章治 中沢洋三
  • Journal Title: 血液内科 Volume: 85 Pages: 814-821
• [Journal Article] Investigation of risk factors associated with erythrocyte engraftment after ABO-incompatible hematopoietic stem cell transplantation (2021)
  • Author(s): Yanagisawa R, Nakazawa H, Nishina S, Saito S, Shigemura T, Tanaka M, Nakazawa Y.
  • Journal Title: Clin Transplant Volume: 27-Mar Issue: 6
  • DOI: 10.1111/ctr.14300
  • Peer Reviewed
• [Journal Article] Autologous non-human primate model for safety assessment of piggyBac transposon-mediated chimeric antigen receptor T cells on granulocyte-macrophage colony-stimulating factor receptor. (2020)
  • Author(s): Morokawa H, Yagyu S, Hasegawa A, Tanaka M, Saito S, Mochizuki H, Sakamoto K, Shimoi A, Nakazawa Y.
  • Journal Title: Clinical & Translational Immunology Volume: 22 Issue: 11
  • DOI: 10.1002/cti2.1207
  • Peer Reviewed / Open Access
• [Journal Article] Assessment of kidney function using inulin-based and estimated glomerular filtration rates before and after allogeneic hematopoietic stem cell transplantation in pediatric patients (2020)
  • Author(s): Matsuoka D, Hirabayashi K, Murase T, Saito S, Hidaka Y, Nakazawa Y.
  • Journal Title: Pediatr Blood Cancer Volume: Dec;67(12) Issue: 12
  • DOI: 10.1002/pbc.28733
  • Peer Reviewed
• [Journal Article] Transfusion outcome for volume- and plasma-reduced platelet concentrates for pediatric patients (2020)
  • Author(s): Tanaka M, Yanagisawa R, Yamanaka M, Konno S, Takemura K, Kojima S, Okura E, Morita D, Saito S, Komori K, Matsuzawa H, Sakashita K, Tozuka M, Nakazawa Y.
  • Journal Title: Transfus Apher Sci Volume: Aug;59(4) Issue: 4 Pages: 102776-102776
  • DOI: 10.1016/j.transci.2020.102776
  • Peer Reviewed
• [Journal Article] Thyroid tumor surveillance using ultrasound in childhood cancer survivors (2020)
  • Author(s): Furui Y, Morita D, Okura E, Komori K, Saito S, Hirabayashi K, Tanaka M, Nakazawa Y.
  • Journal Title: Pediatr Int Volume: May;62(5) Issue: 5 Pages: 562-568
  • DOI: 10.1111/ped.14179
  • Peer Reviewed
• [Journal Article] Comparative analysis of graft-versus-host disease prophylaxis with tacrolimus in combination with methylprednisolone or methotrexate after umbilical cord blood transplantation (2020)
  • Author(s): Shigemura T, Sakashita K, Okura E, Morita D, Komori K, Kurata T, Hirabayashi K, Saito S, Tanaka M, Yanagisawa R, Nakazawa Y. Comparative analysis of graft-versus-host disease prophylaxis with tacrolimus in combination with methylprednisolone or methotrexate after umbilical cord blood transplantation
  • Journal Title: Int J Hematol Volume: 111 Issue: 5 Pages: 702-710
  • DOI: 10.1007/s12185-020-02826-9
  • Peer Reviewed
• [Journal Article] Delayed methotrexate clearance and acute kidney injury after high-dose methotrexate chemotherapy concurrent with dasatinib in a patient with relapsed Philadelphia chromosome-positive acute lymphoblastic leukemia: A case report (2020)
  • Author(s): Morota H, Saito S, Uchida E, Okura E, Matsuoka D, Hidaka Y, Natsume T, Watanabe T, Shigemura T, Morita D, Tanaka M, Kurata T, Sakashita K, Nakazawa Y.
  • Journal Title: Pediatr Blood Cancer Volume: Apr;67(4) Issue: 4
  • DOI: 10.1002/pbc.28173
  • Peer Reviewed
• [Journal Article] Complete measurable residual disease response after combination chemotherapy with AML-type and ALL-type regimens in pediatric B/myeloid acute bilineal leukemia (2020)
  • Author(s): Saito S, Taira C, Matsuda K, Yanagisawa R, Morita D, Shigemura T, Yoshikawa K, Tanaka M, Sakashita K, Mukai S, Shimodaira S, Koike K, Nakazawa Y
  • Journal Title: Leuk Lymphoma Volume: 61 Issue: 4 Pages: 967-970
  • DOI: 10.1080/10428194.2019.1697813
  • Peer Reviewed
• [Presentation] Non-Viral Delivery of Therapeutic mRNA Using Tumor-Tropic Liposomes for the Treatment of Solid Tumors (2021)
  • Author(s): Saito S, Nakashima I, Akahoshi E, Ishihara-Sugano M, Yagyu S, Nakazawa Y.
  • Organizer: American Society of Gene and Cell Therapy 24th Annual meeting
  • Int'l Joint Research
• [Presentation] 難治性血液腫瘍に対する遺伝子・細胞治療の開発 (2021)
  • Author(s): 齋藤章治
  • Organizer: 第124回日本小児科学会学術集会
  • Invited
• [Presentation] Tumor-Tropic Liposome-Based Delivery of Therapeutic mRNA for T Cell Leukemias (2020)
  • Author(s): Saito, S; Nakashima, I; Hasegawa, A; Akahoshi, E; Sugano, M; Yagyu, S; Nakazawa, Y
  • Organizer: 第62回日本小児血液・がん学会
• [Presentation] Tumor-Tropic Liposome-Mediated Therapeutic Delivery of mRNA for T Cell Malignancies (2020)
  • Author(s): Saito, S; Nakashima, I; Hasegawa, A; Akahoshi, E; Ishihara-Sugano, M; Yagyu, S; Nakazawa, Y
  • Organizer: 63rd ASH Annual Meeting & Exposition
  • Int'l Joint Research
• [Presentation] Tumor-Tropic Liposome-Mediated Therapeutic Delivery of mRNA for T Cell Malignancies (2020)
  • Author(s): Saito, S; Nakashima, I; Hasegawa, A; Akahoshi, E; Ishihara-Sugano, M; Yagyu, S; Nakazawa, Y
  • Organizer: 23rd Annual Meeting of the American Society for Gene and Cell Therapy
  • Int'l Joint Research
• [Presentation] Tumor-tropic liposome-mediated therapeutic delivery of mRNA for pediatric / adolescent and young adult cancers. (2020)
  • Author(s): Saito, S; Nakashima, I; Hasegawa, A; Akahoshi, E; Sugano, M; Yagyu, S; Nakazawa, Y
  • Organizer: 第79回日本癌学会学術集会
• [Presentation] 1.Tumor-tropic liposome-mediated therapeutic delivery of mRNA for T cell malignancies (2020)
  • Author(s): Saito, S; Nakashima, I; Hasegawa, A; Akahoshi, E; Sugano, M; Yagyu, S; Nakazawa, Y
  • Organizer: 第82回日本血液学会学術集会