Development of synthetic basement membrane using laminin active peptides

• Project/Area Number: 18K06637
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 47030:Pharmaceutical hygiene and biochemistry-related
• Research Institution: Tokyo University of Pharmacy and Life Science
• Principal Investigator: Nomizu Motoyoshi 東京薬科大学, 薬学部, 教授 (00311522)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2018: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: バイオマテリアル / ペプチド / 細胞接着 / ラミニン / 基底膜 / 細胞外マトリックス / インテグリン / ジストログリカン / αジストログリカン / マトリックス

Outline of Final Research Achievements

Five laminin active peptides that bind to integrins were identified from the laminin α5 chain G domain and β1-3 chain peptide library. We found some laminin active peptides that specifically induced neural differentiation using the peptide-matrix method. We also identified a dystroglycan binding peptide from laminin α5 chain peptide library and the dystroglycan binding peptide was found to promote iPS cell adhesion. We succeeded in developing a heat-responsive peptide matrix using a laminin active peptide and an elastin-like peptide polymer. In addition, cyclized integrin binding laminin peptides were found to enhance the cell attachment activity. We succeeded in a muscle cell specific drug delivery using a dystroglycan binding laminin peptide.

Academic Significance and Societal Importance of the Research Achievements

ラミニン活性ペプチドを用いたペプチド－マトリックスがiPS細胞や神経幹細胞など様々な細胞の培養や分化誘導を目的としたバイオマテリアルとして有用であることを示した。これらの結果は人工基底膜の創製につながるものである。また、ラミニンペプチドを固定化したエラスチン様ペプチドは熱応答性で、バイオマテリアル分野での応用が期待される。環状化によるペプチドの活性増強の結果は、インテグリンとリガンドとの結合メカニズムを解析する上で重要な知見となる。ジストログリカン結合ペプチドによる筋細胞特異的な薬物送達の結果は、ラミニンペプチドが細胞培養用バイオマテリアルだけでなく、広範囲での応用の可能性を示すものである。

Research Products

• [Int'l Joint Research] University of Southern California(米国)
• [Journal Article] Alpha-dystroglycan binding peptide A2G80-modified stealth liposomes as a muscle-targeting carrier for Duchenne muscular dystrophy (2021)
  • Author(s): Sasaki Eri、Hayashi Yoshihiro、Kimura Yuka、Sashida Sanae、Hamano Nobuhito、Nirasawa Kei、Hamada Keisuke、Katagiri Fumihiko、Kikkawa Yamato、Sakai Takaaki、Yoshida Akihiro、Kawada Masahiro、Hirashima Shin-ichi、Miura Tsuyoshi、Endo-Takahashi Yoko、Nomizu Motoyoshi、Negishi Yoichi
  • Journal Title: Journal of Controlled Release Volume: 329 Pages: 1037-1045
  • DOI: 10.1016/j.jconrel.2020.10.032
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Development of A2G80 peptide-gene complex for targeted delivery to muscle cells (2021)
  • Author(s): Nirasawa Kei、Hamada Keisuke、Naraki Yukiko、Kikkawa Yamato、Sasaki Eri、Endo-Takahashi Yoko、Hamano Nobuhito、Katagiri Fumihiko、Nomizu Motoyoshi、Negishi Yoichi
  • Journal Title: Journal of Controlled Release Volume: 329 Pages: 988-996
  • DOI: 10.1016/j.jconrel.2020.10.029
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Evaluation of extracellular matrix mimetic laminin bioactive peptide and elastin‐like polypeptide (2020)
  • Author(s): Truong Anh Tan、Hamada Keisuke、Yamada Yuji、Guo Hao、Kikkawa Yamato、Okamoto Curtis T.、MacKay J. Andrew、Nomizu Motoyoshi
  • Journal Title: The FASEB Journal Volume: 34 Issue: 5 Pages: 6729-6740
  • DOI: 10.1096/fj.201902794r
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Development of Three-Dimensional Cell Culture Scaffolds Using Laminin Peptide-Conjugated Agarose Microgels (2020)
  • Author(s): Yamada Yuji、Yoshida Chihiro、Hamada Keisuke、Kikkawa Yamato、Nomizu Motoyoshi
  • Journal Title: Biomacromolecules Volume: 21 Issue: 9 Pages: 3765-3771
  • DOI: 10.1021/acs.biomac.0c00871
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Conformational dependence of integrin‐binding peptides derived from homologous loop regions in the laminin α chains (2020)
  • Author(s): Ishikawa Masaya、Hamada Keisuke、Yamada Yuji、Kumai Jun、Katagiri Fumihiko、Kikkawa Yamato、Nomizu Motoyoshi
  • Journal Title: Journal of Peptide Science Volume: 26 Issue: 12
  • DOI: 10.1002/psc.3284
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Effects of laminin-111 peptide coatings on rat neural stem/progenitor cell culture (2020)
  • Author(s): Hayashi H, Horinokita I, Yamada Y, Hamada K, Takagi N, Nomizu M.
  • Journal Title: Exp Cell Res Volume: 400 Pages: 112440-112440
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Fibulin-7 C-terminal fragment and its active synthetic peptide suppress choroidal and retinal neovascularization (2020)
  • Author(s): Ikeuchi Tomoko、Kanan Yogita、Long Da、de Vega Susana、Hozumi Kentaro、Nomizu Motoyoshi、Campochiaro Peter A.、Yamada Yoshihiko
  • Journal Title: Microvascular Research Volume: 129 Pages: 103986-103986
  • DOI: 10.1016/j.mvr.2020.103986
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Identification of active sequences in human laminin α5 G domain (2019)
  • Author(s): Kumai Jun、Yamada Yuji、Hamada Keisuke、Katagiri Fumihiko、Hozumi Kentaro、Kikkawa Yamato、Nomizu Motoyoshi
  • Journal Title: Journal of Peptide Science Volume: 25 Issue: 12 Pages: 1-9
  • DOI: 10.1002/psc.3218
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Characterization of dystroglycan binding in adhesion of human induced pluripotent stem cells to laminin-511 E8 fragment (2019)
  • Author(s): Sugawara Yumika、Hamada Keisuke、Yamada Yuji、Kumai Jun、Kanagawa Motoi、Kobayashi Kazuhiro、Toda Tatsushi、Negishi Yoichi、Katagiri Fumihiko、Hozumi Kentaro、Nomizu Motoyoshi、Kikkawa Yamato
  • Journal Title: Scientific Reports Volume: 9 Issue: 1 Pages: 1-12
  • DOI: 10.1038/s41598-019-49669-x
  • NAID: 120006728095
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Development of Antibody-Modified Nanobubbles Using Fc-Region-Binding Polypeptides for Ultrasound Imaging (2019)
  • Author(s): Hamano Nobuhito、Kamoshida Sho、Kikkawa Yamato、Yano Yusuke、Kobayashi Tomomi、Endo-Takahashi Yoko、Suzuki Ryo、Maruyama Kazuo、Ito Yuji、Nomizu Motoyoshi、Negishi Yoichi
  • Journal Title: Pharmaceutics Volume: 11 Issue: 6 Pages: 283-283
  • DOI: 10.3390/pharmaceutics11060283
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Laminin-derived peptides: Applications in drug delivery systems for targeting (2019)
  • Author(s): Negishi Yoichi、Nomizu Motoyoshi
  • Journal Title: Pharmacology & Therapeutics Volume: 202 Pages: 91-97
  • DOI: 10.1016/j.pharmthera.2019.05.017
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Identification of specific integrin cross-talk for dermal fibroblast cell adhesion using a mixed peptide-chitosan matrix (2019)
  • Author(s): Hozumi Kentaro、Teranishi Yui、Enomoto Sayaka、Katagiri Fumihiko、Kikkawa Yamato、Nomizu Motoyoshi
  • Journal Title: Journal of Biomaterials Applications Volume: 33 Issue: 7 Pages: 893-902
  • DOI: 10.1177/0885328218823457
  • Peer Reviewed / Open Access
• [Journal Article] Mixed Peptide-Conjugated Chitosan Matrices as Multi-Receptor Targeted Cell-Adhesive Scaffolds (2018)
  • Author(s): Hozumi K, Nomizu M
  • Journal Title: Int J Mol Sci. Volume: 19(9) Issue: 9 Pages: 2713-2713
  • DOI: 10.3390/ijms19092713
  • Peer Reviewed / Open Access
• [Journal Article] Cell adhesion activity of peptides conjugated to polysaccharides (2018)
  • Author(s): Hozumi K, Nomizu M
  • Journal Title: Curr Protoc Cell Biol. Volume: 80(1) Issue: 1
  • DOI: 10.1002/cpcb.53
  • Peer Reviewed / Open Access
• [Presentation] Peptide-chitosan Matrix for Cell Adhesive Biomaterials (2021)
  • Author(s): Motoyoshi Nomizu
  • Organizer: The 18th Akabori Conference (German-Japanese Symposium on Peptide Science)
  • Int'l Joint Research
• [Presentation] ラミニンα2鎖LG4-5モジュール由来α-ジストログリカン結合ペプチドの同定とその構造活性相関研究 (2020)
  • Author(s): 濵田圭佑、張光端、山田雄二、吉川大和、野水基義
  • Organizer: 第52回 日本結合組織学会学術大会
• [Presentation] ジカルボン酸で架橋したキトサンハイドロゲルを用いた細胞培養プラットフォームの開発 (2020)
  • Author(s): 保住建太郎、髙橋知衣、濵田圭佑、山田雄二、吉川大和、野水基義
  • Organizer: 第52回 日本結合組織学会学術大会
• [Presentation] ラミニンペプチド修飾アガロースゲルを用いた三次元培養システムの開発 (2020)
  • Author(s): 山田雄二、吉田智浩、濵田圭佑、野水基義、吉川大和
  • Organizer: 第52回 日本結合組織学会学術大会
• [Presentation] Laminin peptide coatings for rat neural stem/progenitor cell culture (2020)
  • Author(s): K. Hamada, H. Hayashi, I. Horinokita, Y. Yamada, Y. Kikkawa, N. Takagi, and M. Nomizu
  • Organizer: 第57回 ペプチド討論会
• [Presentation] Three-dimensional cell culture using laminin peptide-conjugated agarose microgels (2020)
  • Author(s): Y. Yamada,C. Yoshida, K. Hamada, Y. Kikkawa, and M. Nomizu
  • Organizer: 第57回 ペプチド討論会
• [Presentation] ラミニン-511E8領域由来のジストログリカン(DG)結合ペプチドに対するヒトiPS細胞の接着 (2019)
  • Author(s): 吉川大和、菅原由美香、濱田圭佑、山田雄二、熊井準、金川基、小林千浩、戸田達史、根岸洋一、片桐文彦、保住健太郎、野水基義
  • Organizer: 第51回 日本結合組織学会学術大会
• [Presentation] ラミニンa2鎖LG4-5モジュールに着目したa-ジストログリカン結合ペプチドの探索 (2019)
  • Author(s): 濵田圭佑、張光端、山田雄二、吉川大和、野水基義
  • Organizer: 第22回 生命化学研究会
• [Presentation] Cell adhesion research based on peptide science (2019)
  • Author(s): 野水基義
  • Organizer: 第56回 ペプチド討論会
  • Invited
• [Presentation] Structure activity relationship study of an a-dystroglycan binding peptide from the laminin a2 chain LG4-5 modules (Chemical Communication Poster Award) (2019)
  • Author(s): 濵田圭佑、張光端、山田雄二、吉川大和、野水基義
  • Organizer: 第56回 ペプチド討論会
• [Presentation] Identification and structure activity relationship study of an a-Dystroglycan binding peptide from the laminin a2 chain LG4-5 modules (2019)
  • Author(s): K. Hamada, G. Zhang, J. Kumai, F. Katagiri, K. Hozumi, Y. Kikkawa, and M. Nomizu
  • Organizer: American Society of Cell Biology/Europian Molecular Biology Organization 2019 meeting
  • Int'l Joint Research
• [Presentation] Characterization of dystroglycan binding in adhesion of human induced pluripotent stem cells to laminin-511 E8 fragment (2019)
  • Author(s): 吉川大和、菅原由美香、濱田圭佑、山田雄二、熊井準、金川基、小林千浩、戸田達史、根岸洋一、片桐文彦、保住健太郎、野水基義
  • Organizer: American Society of Cell Biology/Europian Molecular Biology Organization 2019 meeting
  • Int'l Joint Research
• [Presentation] Modification of chain-specific cell adhesion sequences in the short arm region of laminin β chain (2018)
  • Author(s): K. Hamada, Y. Gu, J. Kumai, K. Nakamura,F. Katagiri, K. Hozumi, Y. Kikkawa, M. Nomizu
  • Organizer: 10th International Peptide Symposium 第55回ペプチド討論会
  • Int'l Joint Research
• [Presentation] Identification of α-dystroglycan binding sequences in the laminin α2 chain LG4-5 modules using the peptide-chitosan matrix (2018)
  • Author(s): G. Zhang, K. Hamada, J. Kumai, F. Katagiri, K. Hozumi, Y. Kikkawa, M. Nomizu
  • Organizer: 10th International Peptide Symposium 第55回ペプチド討論会
  • Int'l Joint Research
• [Presentation] Identification of alpha-dystroglycan bindingpeptide sequences in the laminin alpha2 chain LG4-5 modules using the peptides-chitosan matrix (2018)
  • Author(s): K. Hamada, G. Zhang, J. Kumai, F. Ktagiri, K. Hozumi, Y. Kikkawa, M. Nomizu
  • Organizer: American Society of Cell Biology/Europian Molecular Biology Organization
  • Int'l Joint Research