Development of immunostimulatory mRNA and lymphoid tissue-targeting carriers for cancer vaccine

• Project/Area Number: 18H03529
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 90120:Biomaterials-related
• Research Institution: Kyoto Prefectural University of Medicine (2020-2021); The University of Tokyo (2018-2019)
• Principal Investigator: 内田 智士 京都府立医科大学, 医学(系)研究科(研究院), 准教授 (20710726)
• Co-Investigator(Kenkyū-buntansha): 弓場 英司 大阪府立大学, 工学(系)研究科(研究院), 准教授 (80582296)
• Project Period (FY): 2018-04-01 – 2022-03-31
• Project Status: Declined (Fiscal Year 2021)
• Budget Amount: ¥17,160,000 (Direct Cost: ¥13,200,000、Indirect Cost: ¥3,960,000); Fiscal Year 2021: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2020: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2019: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2018: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
• Keywords: mRNAワクチン / がんワクチン / 細胞性免疫 / 2本鎖RNA / 脂質性ナノ粒子 / 高分子ミセル / アジュバント / ガンワクチン / 2本鎖mRNA

Outline of Annual Research Achievements

mRNAワクチンは効果的に細胞性免疫を誘導できるため、がんワクチンへ有望である。mRNAワクチンでは、主に脂質性ナノ粒子を構成する脂質性ナノ粒子を構成する脂質に免疫賦活化アジュバントとしての活性が付与されてきたが、脂質は体内動態の制御が困難であり、我々はより動態制御が容易なmRNAにアジュバント活性を付与した。本研究では、mRNAへのアジュバント活性の導入方法と、mRNA送達システムの構築を行っている。これまでに、mRNAに2本鎖構造を搭載することで、アジュバントとしての活性を併せ持つ設計を最適化し、さらに、免疫原性の低い脂質性ナノ粒子と併用することで、優れたワクチン効果が得られることを、マウスにて実証していた。
本年度の研究では、モデル抗原を発現するがん細胞を移植したマウスモデルにおいて、前年度までに構築したmRNAワクチンシステムにより、高い抗がん活性が得られることを示した。すなわち、独自に設計したシステムのがんワクチンへの有効性が実証された。
また、より安全性の高い投与を目指し、脂質を用いないmRNA送達方法も検討している。これまでに、我々は表面がポリエチレングリコールで覆われた高分子ミセルを用いることで、安全なmRNA送達が可能であることを示してきた。一方、この高分子ミセルは、安全性が高いものの、RNA分解酵素に対する耐性が低いことが課題であった。これに対して、本年度、mRNA修飾など様々な方法論でこの課題を克服することに成功した。さらに、高分子ミセルを用いて独自の2本鎖構造を有するmRNを送達することで、マウスに対して優れた細胞性免疫誘導活性を得ることができることも実証した。
以上のように、本年度までに、独自に開発したシステムが、モデル抗原を用いた評価において、優れた効果を示すことを実証した。

Research Progress Status

翌年度、交付申請を辞退するため、記入しない。

Strategy for Future Research Activity

翌年度、交付申請を辞退するため、記入しない。

Research Products

• [Journal Article] Co-encapsulation of Cas9 mRNA and guide RNA in polyplex micelles enables genome editing in mouse brain (2021)
  • Author(s): S. Abbasi, S. Uchida, K. Toh, T.A. Tockary, A. Dirisala, K. Hayashi, S. Fukushima, K. Kataoka
  • Journal Title: Journal of Controlled Release Volume: 332 Pages: 260-268
  • DOI: 10.1016/j.jconrel.2021.02.026
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Treatment of ischemic neuronal death by introducing brain-derived neurotrophic factor mRNA using polyplex nanomicelle (2021)
  • Author(s): Y. Fukushima, S. Uchida, H. Imai, H. Nakatomi, K. Kataoka, N. Saito, K. Itaka
  • Journal Title: Biomaterials Volume: 270 Pages: 120681-120681
  • DOI: 10.1016/j.biomaterials.2021.120681
  • Peer Reviewed
• [Journal Article] mRNA loading into ATP-responsive polyplex micelles with optimal density of phenylboronate ester crosslinking to balance robustness in the biological milieu and intracellular translational efficiency (2021)
  • Author(s): Yoshinaga Naoto、Uchida Satoshi、Dirisala Anjaneyulu、Naito Mitsuru、Osada Kensuke、Cabral Horacio、Kataoka Kazunori
  • Journal Title: J. Control. Release Volume: 330 Pages: 317-328
  • DOI: 10.1016/j.jconrel.2020.12.033
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Transient stealth coating of liver sinusoidal wall by anchoring two-armed PEG for retargeting nanomedicines (2020)
  • Author(s): Dirisala Anjaneyulu、Uchida Satoshi、Toh Kazuko、Li Junjie、Osawa Shigehito、Tockary Theofilus A.、Liu Xueying、Abbasi Saed、Hayashi Kotaro、Mochida Yuki、Fukushima Shigeto、Kinoh Hiroaki、Osada Kensuke、Kataoka Kazunori
  • Journal Title: Science Advances_ Volume: 6
  • DOI: 10.1126/sciadv.abb8133
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Bundling of mRNA strands inside polyion complexes improves mRNA delivery efficiency in vitro and in vivo (2020)
  • Author(s): Koji Kyoko、Yoshinaga Naoto、Mochida Yuki、Hong Taehun、Miyazaki Takuya、Kataoka Kazunori、Osada Kensuke、Cabral Horacio、Uchida Satoshi
  • Journal Title: Biomaterials Volume: 261 Pages: 120332-120332
  • DOI: 10.1016/j.biomaterials.2020.120332
  • Peer Reviewed
• [Journal Article] Guanidine-phosphate interactions stabilize polyion complex micelles based on flexible catiomers to improve mRNA delivery (2020)
  • Author(s): T. Miyazaki, S. Uchida, H. Hatano, Y. Miyahara, A. Matsumoto, H. Cabral
  • Journal Title: European Polymer Journal Volume: 140 Pages: 110028-110028
  • DOI: 10.1016/j.eurpolymj.2020.110028
  • Peer Reviewed
• [Journal Article] Polymeric Nanocarriers with Controlled Chain Flexibility Boost mRNA Delivery In Vivo through Enhanced Structural Fastening (2020)
  • Author(s): Miyazaki Takuya、Uchida Satoshi、Nagatoishi Satoru、Koji Kyoko、Hong Taehun、Fukushima Shigeto、Tsumoto Kouhei、Ishihara Kazuhiko、Kataoka Kazunori、Cabral Horacio
  • Journal Title: Adv Healthc Mater Volume: 9 Pages: 2000538-2000538
  • DOI: 10.1002/adhm.202000538
  • Peer Reviewed
• [Journal Article] Nanomedicine-Based Approaches for mRNA Delivery (2020)
  • Author(s): S. Uchida, F. Perche, C. Pichon, H. Cabral
  • Journal Title: Mol. Pharm._ Volume: 17 Pages: 3654-3684
  • DOI: 10.1021/acs.molpharmaceut.0c00618
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] mRNA as a Tool for Gene Transfection in 3D Cell Culture for Future Regenerative Therapy (2020)
  • Author(s): S. Uchida, K. Yanagihara, A. Matsui, K. Kataoka, K. Itaka
  • Journal Title: Micromachines Volume: 11 Pages: 426-426
  • DOI: 10.3390/mi11040426
  • Peer Reviewed / Open Access
• [Journal Article] A chemically unmodified agonistic DNA with growth factor functionality for in vivo therapeutic application (2020)
  • Author(s): Ueki Ryosuke、Uchida Satoshi、Kanda Naoto、Yamada Naoki、Ueki Ayaka、Akiyama Momoko、Toh Kazuko、Cabral Horacio、Sando Shinsuke
  • Journal Title: Science Advances_ Volume: 6
  • DOI: 10.1126/sciadv.aay2801
  • Peer Reviewed / Open Access
• [Journal Article] A 50-nm-Sized Micellar Assembly of Thermoresponsive Polymer-Antisense Oligonucleotide Conjugates for Enhanced Gene Knockdown in Lung Cancer by Intratracheal Administration. (2020)
  • Author(s): B. S. Kim, S. Osawa, M. Naito, S. Ogura, R. Kamegawa, H. Ishida, H. J. Kim, S. Uchida, K. Miyata
  • Journal Title: Advanced Therapeutics Volume: 3 Pages: 1900123-1900123
  • DOI: 10.1002/adtp.201900123
  • Peer Reviewed
• [Journal Article] Single-stranded DNA-packaged polyplex micelle as adeno-associated-virus-inspired compact vector to systemically target stroma-rich pancreatic cancer. (2019)
  • Author(s): T. A. Tockary, W. Foo, A. Dirisala, Q. Chen, S. Uchida, S. Osawa, Y. Mochida, X. Liu, H. Kinoh, H. Cabral, K. Osada, K. Kataoka
  • Journal Title: ACS Nano Volume: 13 Pages: 12732-12742
  • DOI: 10.1021/acsnano.9b04676
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Enhancement of motor function 1 recovery after spinal cord injury in mice by delivery of brain derived neurotrophic factor mRNA. (2019)
  • Author(s): S. T. Crowley, Y. Fukushima, S. Uchida, K. Kataoka, K. Itaka
  • Journal Title: Mol Ther Nucleic Acids Volume: 17 Pages: 465-476
  • DOI: 10.1016/j.omtn.2019.06.016
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Bundling mRNA strands to prepare nano-assemblies with enhanced stability towards RNase for in vivo delivery. (2019)
  • Author(s): N. Yoshinaga, E. Cho, K. Koji, Y. Mochida, M. Naito, K. Osada, K. Kataoka, H. Cabral, S. Uchida
  • Journal Title: Angewandte Chemie International Edition Volume: 58 Pages: 11360-11363
  • DOI: 10.1002/anie.201905203
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] In vivo rendezvous of small nucleic acid drugs with charge-matched block catiomers to target cancers (2019)
  • Author(s): Watanabe Sumiyo、Hayashi Kotaro、Toh Kazuko、Kim Hyun Jin、Liu Xueying、Chaya Hiroyuki、Fukushima Shigeto、Katsushima Keisuke、Kondo Yutaka、Uchida Satoshi、Ogura Satomi、Nomoto Takahiro、et al.
  • Journal Title: Nat. Commun. Volume: 10 Pages: 1-13
  • DOI: 10.1038/s41467-019-09856-w
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] PEG-OligoRNA Hybridization of mRNA for Developing Sterically Stable Lipid Nanoparticles toward In Vivo Administration (2019)
  • Author(s): Kurimoto Shota、Yoshinaga Naoto、Igarashi Kazunori、Matsumoto Yu、Cabral Horacio、Uchida Satoshi
  • Journal Title: Molecules Volume: 24 Pages: 1303-1303
  • DOI: 10.3390/molecules24071303
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Treatment of intervertebral disk disease by administration of messenger RNA encoding a cartilage-anabolic transcription factor. (2019)
  • Author(s): C. -Y. Lin, S. T. Crowley, S. Uchida, Y. Komaki, K. Kataoka, K. Itaka
  • Journal Title: Molecular Therapy - Nucleic Acids Volume: 16 Pages: 162-171
  • DOI: 10.1016/j.omtn.2019.02.012
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Design concepts of polyplex micelles for in vivo therapeutic delivery of plasmid DNA and messenger RNA. (2019)
  • Author(s): S. Uchida, K. Kataoka
  • Journal Title: Journal of Biomedical Materials Research Part A Volume: 107 Pages: 978-990
  • DOI: 10.1002/jbm.a.36614
  • Peer Reviewed
• [Journal Article] Precise tuning of disulfide crosslinking in mRNA polyplex micelles for optimizing extracellular and intracellular nuclease tolerability. (2019)
  • Author(s): A. Dirisala, S. Uchida, T. A. Tockary, N. Yoshinaga, J. Li, S. Osawa, L. Gorantla, S. Fukushima, K. Osada, K. Kataoka
  • Journal Title: Journal of Drug Targeting Volume: 印刷中 Pages: 670-680
  • DOI: 10.1080/1061186x.2018.1550646
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Tunable non-enzymatic degradability of N-substituted polyaspartamide main chain by amine protonation and alkyl spacer length in side chains for enhanced messenger RNA transfection efficiency. (2019)
  • Author(s): M. Naito, Y. Otsu, R. Kamegawa, K. Hayashi, S. Uchida, H. -J. Kim, K. Miyata
  • Journal Title: Sci. Technol. Adv. Mater. Volume: 20 Pages: 105-115
  • DOI: 10.1080/14686996.2019.1569818
  • Peer Reviewed / Open Access
• [Journal Article] Induced packaging of mRNA into polyplex micelles by regulated hybridization with a small number of cholesteryl RNA oligonucleotides directed enhanced in vivo transfection. (2019)
  • Author(s): N. Yoshinaga, S. Uchida, M. Naito, K. Osada, H. Cabral, K. Kataoka
  • Journal Title: Biomaterials Volume: 197 Pages: 255-267
  • DOI: 10.1016/j.biomaterials.2019.01.023
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Prolonged engraftment of transplanted hepatocytes in the liver by transient pro-survival factor supplementation using ex vivo mRNA transfection. (2018)
  • Author(s): A. Matsui, S. Uchida, A. Hayashi, K. Kataoka, K. Itaka
  • Journal Title: J Control Release Volume: 285 Pages: 1-11
  • DOI: 10.1016/j.jconrel.2018.06.033
  • Peer Reviewed / Open Access
• [Presentation] メッセンジャーRNAのCOVID-19ワクチンへの応用とその後の展望 (2021)
  • Author(s): 内田 智士
  • Organizer: 2020年度 医療産業イノベーションフォーラム，
  • Invited
• [Presentation] 新型コロナウイルスや未来のパンデミックに対するmRNAナノワクチンの開発 (2020)
  • Author(s): 内田 智士
  • Organizer: 第7回COINSシンポジウム ～小さく寄り添い大きく守る、未来の医療はすぐそこに～
  • Invited
• [Presentation] RNAオリゴマーを用いたmRNA修飾法の開発とmRNA送達、ワクチンへの展開 (2020)
  • Author(s): 内田 智士
  • Organizer: 核酸医薬シンポジウム2020
  • Invited
• [Presentation] mRNA bundling inside polyion complexes for their physical and biological stabilization (2020)
  • Author(s): S. Uchida, K. Koji, N. Yoshinaga, Y. Mochida, T. Hong, H. Cabral
  • Organizer: 8th International mRNA Health Conference
  • Int'l Joint Research
• [Presentation] mRNA nanomachine for treating rare genetic disorders (2020)
  • Author(s): S. Uchida
  • Organizer: India-Japan Webinar on "Rare Genetic Disorders"
  • Int'l Joint Research / Invited
• [Presentation] RNA構造に着目したメッセンジャーRNA 医薬の機能化 (2020)
  • Author(s): 内田 智士
  • Organizer: 第69回 高分子討論会
  • Invited
• [Presentation] DNA、mRNAのex vivo導入による細胞治療の効果向上 (2020)
  • Author(s): 内田 智士
  • Organizer: 第36回日本DDS学会学術集会
  • Invited
• [Presentation] Preparation of nanoassemblies from several mRNA strands for enhancing nuclease stability. (2020)
  • Author(s): S. Uchida, N. Yoshinaga, E. Cho, K. Koji, K. Kataoka, H. Cabral
  • Organizer: International Symposium on Biomedical Materials for Drug/Gene Delivery
  • Int'l Joint Research
• [Presentation] Development of mRNA polyplex micelles through engineering polymers and mRNA for in vivo therapeutic applications (2020)
  • Author(s): S. Uchida
  • Organizer: IBS CNM Seminar
  • Int'l Joint Research / Invited
• [Presentation] Stabilization of mRNA polyplex micelles for in vivo delivery by engineering mRNA (2019)
  • Author(s): S. Uchida, N. Yoshinaga, H. Cabral, K. Kataoka
  • Organizer: 2nd G’L’owing Polymer Symposium in KANTO
• [Presentation] 複複数のmRNA鎖を束ねることで生体内酵素分解を抑制させたmRNAナノ集合体の開発 (2019)
  • Author(s): 内田 智士、吉永 直人、趙 オル、小路 恭子、長田 健介、片岡 一則、Horacio Cabral
  • Organizer: 第41回バイオマテリアル学会大会
• [Presentation] Nanoengineering mRNA through short RNA hybridization for biomedical application (2019)
  • Author(s): S. Uchida
  • Organizer: 1st Thailand-Australia-Japan Joint Research Meeting on Nanomedicine
  • Int'l Joint Research / Invited
• [Presentation] 複数のmRNA鎖を束ねたナノ集合体の開発と生体内投与 (2019)
  • Author(s): 内田 智士、吉永 直人、趙 オル、小路 恭子、長田 健介、片岡 一則、Horacio Cabral
  • Organizer: 日本核酸医薬学会第5回年会
• [Presentation] 複数のmRNA鎖からなるナノ集合体を用いたmRNA安定化とその生体内投与 (2019)
  • Author(s): 内田 智士、吉永 直人、趙 オル、小路 恭子、長田 健介、片岡 一則、Horacio Cabral
  • Organizer: 第35回日本DDS学会学術集会
• [Presentation] Hybridization of mRNA with a small number of short cholesteryl RNA oligonucleotides for tight mRNA packaging to polyplex micelles. (2019)
  • Author(s): S. Uchida, N. Yoshinaga, H. Cabral, K. Kataoka
  • Organizer: 19th Smposium for gene design and delivery
  • Int'l Joint Research
• [Presentation] コレステロール修飾相補鎖RNAを用いたmRNA搭載高分子ミセル安定化 (2018)
  • Author(s): 内田 智士、吉永 直人、長田 健介、位髙 啓史、Horacio Cabral、片岡 一則
  • Organizer: 第40回日本バイオマテリアル学会大会
• [Presentation] 免疫賦活化アジュバント機能一体型 2本鎖mRNAワクチン (2018)
  • Author(s): 内田 智士、位髙 啓史、片岡 一則
  • Organizer: 日本核酸医薬学会第4回年会
• [Presentation] 高分子ミセルを用いた メッセンジャー(m)RNA医薬の送達と 疾患治療への展開 (2018)
  • Author(s): 内田 智士
  • Organizer: 第34回日本ＤＤＳ学会学術集会
• [Presentation] Therapeutic application of mRNA delivery using polyplex micelles (2018)
  • Author(s): Satoshi Uchida, Keiji Itaka, Kazunori Kataoka
  • Organizer: Finland-Japan Workshop: The next generation medical engineering in biomaterials
  • Int'l Joint Research
• [Presentation] Incorporation of Immunostimulatory Property to Messenger RNA Molecule by Hybridizing RNA to Poly A Sequence for Effective Vaccination (2018)
  • Author(s): Satoshi Uchida, Keiji Itaka, Kazunori Kataoka
  • Organizer: 21st annual meeting ASGCT
  • Int'l Joint Research
• [Remarks] ホームページ
  • URL: http://www.f.kpu-m.ac.jp/y/medchem/uchida.html
• [Remarks] ホームページ
  • URL: http://www.bmc.t.u-tokyo.ac.jp/suchida.html
• [Remarks]
  • URL: http://www.bmc.t.u-tokyo.ac.jp/suchida.html
• [Patent(Industrial Property Rights)] ゲノム編集用ミセル (2020)
  • Inventor(s): 1.片岡 一則，内田 智士，アッバシ サエド
  • Industrial Property Rights Holder: 川崎市産業振興財団
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2020-113434
  • Filing Date: 2020
• [Patent(Industrial Property Rights)] mRNA送達用キャリアおよびこれを含む組成物 (2019)
  • Inventor(s): 片岡 一則，内田 智士，アンジャネユル ディリサラ
  • Industrial Property Rights Holder: 川崎市産業振興財団
  • Industrial Property Rights Type: 特許
  • Filing Date: 2019
  • Overseas