Determination of therapeutic factors using extracellular vesicles secreted from 3D-cultured stem cells

• Project/Area Number: 17K20086
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Research Field: Biomedical engineering and related fields
• Research Institution: Kyoto Prefectural University of Medicine (2020); The University of Tokyo (2017-2019)
• Principal Investigator: Uchida Satoshi 京都府立医科大学, 医学(系)研究科(研究院), 准教授 (20710726)
• Project Period (FY): 2017-06-30 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000); Fiscal Year 2019: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2018: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2017: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: 細胞外小胞 / 3次元培養 / 細胞移植治療 / 再生医療 / 核酸医薬 / スフェロイド / 間葉系幹細胞 / 炎症 / miRNA / 骨再生治療 / 再生医学 / ナノバイオ / 薬学

Outline of Final Research Achievements

In cell transplantation therapy, extracellular vesicles (EVs) secreted by the cells play significant therapeutic roles. This motivated us to study EVs for determining potent therapeutic factors. We focused on EVs secreted from MSCs in three-dimensional culture, which showed enhanced therapeutic effect compared to MSCs in conventional two-dimensional culture. Comprehensive comparison between EVs secreted from MSCs in two- and three-dimensional culture allowed us to determine various microRNAs possessing therapeutic functionalities, including anti-inflammatory and osteogenetic effects.

Academic Significance and Societal Importance of the Research Achievements

本研究の成果は、細胞移植治療のメカニズム解明に繋がるだけでなく、細胞を使わない再生医療の実現に資する。細胞移植は優れた治療効果を示すものの、経済的コストや安全性に課題を残す。今回、移植細胞が分泌するEVに含まれる、高い治療効果を持つmiRNAを同定することに成功した。このmiRNAを治療薬として用いることで、細胞移植を行うことなく、同等の治療効果が得られる可能性がある。このことで、これまで重篤な疾患が主な対象であった再生医療を、より幅広い疾患へと応用できることが期待される。

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: J. Control. 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: Journal of Controlled 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 Issue: 26
  • 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 / Int'l Joint Research
• [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: Eur. Polym. J. Volume: 140 Pages: 110028-110028
  • DOI: 10.1016/j.eurpolymj.2020.110028
  • Peer Reviewed / Int'l Joint Research
• [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: Advanced Healthcare Materials Volume: 9 Issue: 16 Pages: 2000538-2000538
  • DOI: 10.1002/adhm.202000538
  • Peer Reviewed / Int'l Joint Research
• [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 Issue: 10 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 Issue: 4 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 Issue: 14
  • 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 Issue: 2 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 Issue: 11 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: Angew. Chem. Int. Ed. Volume: 58 Issue: 33 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: Nature Communications Volume: 10 Issue: 1 Pages: 1894-1894
  • 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 Issue: 7 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: Mol. Ther. 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: J. Biomed. Mater. Res. A Volume: 107 Issue: 5 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: J. Drug Target. Volume: 印刷中 Issue: 5-6 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 Issue: 1 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
• [Journal Article] Treatment of Bone Defects by Transplantation of Genetically Modified Mesenchymal Stem Cell Spheroids (2018)
  • Author(s): Yanagihara K, Uchida S, Ohba S, Kataoka K, Itaka K
  • Journal Title: Molecular Therapy - Methods & Clinical Development Volume: 9 Pages: 358-366
  • DOI: 10.1016/j.omtm.2018.04.006
  • Peer Reviewed / Open Access
• [Journal Article] Designing immunostimulatory double stranded messenger RNA with maintained translational activity through hybridization with poly A sequences for effective vaccination (2018)
  • Author(s): Uchida Satoshi、Yoshinaga Naoto、Yanagihara Kayoko、Yuba Eiji、Kataoka Kazunori、Itaka Keiji
  • Journal Title: Biomaterials Volume: 150 Pages: 162-170
  • DOI: 10.1016/j.biomaterials.2017.09.033
  • Peer Reviewed
• [Journal Article] Polyplex Micelles with Phenylboronate/Gluconamide Cross-Linking in the Core Exerting Promoted Gene Transfection through Spatiotemporal Responsivity to Intracellular pH and ATP Concentration (2017)
  • Author(s): Yoshinaga Naoto、Ishii Takehiko、Naito Mitsuru、Endo Taisuke、Uchida Satoshi、Cabral Horacio、Osada Kensuke、Kataoka Kazunori
  • Journal Title: Journal of the American Chemical Society Volume: 139 Issue: 51 Pages: 18567-18575
  • DOI: 10.1021/jacs.7b08816
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Glycaemic control boosts glucosylated nanocarrier crossing the BBB into the brain (2017)
  • Author(s): Anraku Y.、Kuwahara H.、Fukusato Y.、Mizoguchi A.、Ishii T.、Nitta K.、Matsumoto Y.、Toh K.、Miyata K.、Uchida S.、Nishina K.、Osada K.、Itaka K.、Nishiyama N.、Mizusawa H.、Yamasoba T.、Yokota T.、Kataoka K.
  • Journal Title: Nat Commun Volume: 8 Issue: 1 Pages: 2001-2009
  • DOI: 10.1038/s41467-017-00952-3
  • Peer Reviewed / Open Access
• [Journal Article] Systemic Delivery of Folate-PEG siRNA Lipopolyplexes with Enhanced Intracellular Stability for In Vivo Gene Silencing in Leukemia (2017)
  • Author(s): Lee Dian-Jang、Kessel Eva、Lehto Taavi、Liu Xueying、Yoshinaga Naoto、Padari K?rt、Chen Ying-Chen、Kempter Susanne、Uchida Satoshi、R?dler Joachim O.、Pooga Margus、Sheu Ming-Thau、Kataoka Kazunori、Wagner Ernst
  • Journal Title: Bioconjugate chemistry Volume: 28 Issue: 9 Pages: 2393-2409
  • DOI: 10.1021/acs.bioconjchem.7b00383
  • Peer Reviewed / Int'l Joint Research
• [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] 標題 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
• [Presentation] 高分子ミセルを用いたメッセンジャーRNAデリバリー (2017)
  • Author(s): 内田智士
  • Organizer: 理研シンポジウム：理研/iCONM/物材研 医工学ネットワーク
  • Invited
• [Presentation] 免疫賦活化能を向上させた2本鎖メッセンジャーRNAワクチンの開発 (2017)
  • Author(s): 内田 智士、吉永 直人、柳原 歌代子、弓場 獎司、位髙 啓史、片岡 一則
  • Organizer: 第39回日本バイオマテリアル学会大会
• [Presentation] mRNA搭載ナノミセルを用いた抗アミロイドβ単鎖抗体導入によるアルツハイマー病治療 (2017)
  • Author(s): 内田 智士、F. Perche、秋葉 宏樹、位髙 啓史、津本 浩平、片岡 一則
  • Organizer: 日本核酸医薬学会第3回年会
• [Presentation] 抗アミロイドβ単鎖抗体発現mRNA搭載ナノミセルを用いたアルツハイマー病治療 (2017)
  • Author(s): 内田 智士、F. Perche、秋葉 宏樹、位髙 啓史、津本 浩平、片岡 一則
  • Organizer: 第33回日本DDS学会学術集会
• [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
• [Patent(Industrial Property Rights)] ゲノム編集用ミセル (2020)
  • Inventor(s): 片岡 一則，内田 智士，アッバシ サエド
  • Industrial Property Rights Holder: 片岡 一則，内田 智士，アッバシ サエド
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2020-113434
  • Filing Date: 2020