Development of functional photosensitizing peptide for photocontrol of biofunctions

• Project/Area Number: 18H02090
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 37010:Bio-related chemistry
• Research Institution: Okayama University
• Principal Investigator: Ohtsuki Takashi 岡山大学, ヘルスシステム統合科学学域, 教授 (80321735)
• Co-Investigator(Kenkyū-buntansha): 舟橋 弘晃 岡山大学, 環境生命科学学域, 教授 (50284089)
• Project Period (FY): 2018-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥17,160,000 (Direct Cost: ¥13,200,000、Indirect Cost: ¥3,960,000); Fiscal Year 2021: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000); Fiscal Year 2020: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2019: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2018: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
• Keywords: 光増感剤 / ペプチド / PCI / Fucci-PCI / 光制御 / 発生 / オプトバイオ

Outline of Final Research Achievements

In this study, we developed a molecular tool combining a photosensitizer and a peptide/protein (functional photosensitizing peptide; FPP) to establish a technology for the spatiotemporal control and observation of cellular functions by light. First, we elucidated the effects of the amino acid sequence near the photosensitizers to support FPP design. Furthermore, by using this technology, we established a method for light-directed cytosolic delivery of two different RNAs, clarified the cell cycle dependence of apoptosis photo-triggered using a FPP, and succeeded in photoinduced RNA delivery into a single cell in mouse embryos.

Academic Significance and Societal Importance of the Research Achievements

光増感剤近傍配列の影響の解明結果は、生命科学研究や光線力学療法のツールとなる幅広い標的機能を持つFPPの設計に役立つ。２波長の光による２種類のRNAの一過的導入法は、複数のRNAによる様々な細胞機能の誘導に応用可能である。Fucci-PCI法はペプチド/タンパク質機能の細胞周期依存性を解析するための有望なツールになると考えられる。マウス卵細胞および初期胚における細胞質内FPP導入の時空間制御法については、哺乳動物の初期発生研究への貢献が期待される。

Research Products

• [Journal Article] Photocontrolled apoptosis induction using precursor miR-664a and an RNA carrier-conjugated with photosensitizer (2021)
  • Author(s): Watanabe, K., Nawachi, T., Okutani, R., Ohtsuki, T.
  • Journal Title: Scientific Reports Volume: 11 Issue: 1 Pages: 14936-14936
  • DOI: 10.1038/s41598-021-94249-7
  • NAID: 120007139720
  • Peer Reviewed / Open Access
• [Journal Article] Fluorophore-PNA-Quencher/Quencher-DNA Probe for miRNA Detection (2021)
  • Author(s): Tabara, K., Watanabe, K., Shigeto, H., Yamamura, S., Kishi, T., Kitamatsu, M., Ohtsuki, T.
  • Journal Title: Bioorganic & Medicinal Chemistry Letters Volume: 51 Pages: 128359-128359
  • DOI: 10.1016/j.bmcl.2021.128359
  • Peer Reviewed
• [Journal Article] Minimization of apoptosis-inducing CPP-Bim peptide. (2021)
  • Author(s): Zhou, S., Watanabe, K., Koide, S., Kitamatsu, M. and Ohtsuki, T.
  • Journal Title: Bioorganic & Medicinal Chemistry Letters Volume: 36 Pages: 127811-127811
  • DOI: 10.1016/j.bmcl.2021.127811
  • Peer Reviewed
• [Journal Article] Lactosome-conjugated siRNA nanoparticles for photo-enhanced gene silencing in cancer cells. (2021)
  • Author(s): Lim, M. S. H., Nishiyama, Y., Ohtsuki, T., Watanabe, K., Kobuchi, H., Kobayashi, K., Matsuura, E.
  • Journal Title: J. Pharm. Sci. Volume: 110 Issue: 4 Pages: 1788-1798
  • DOI: 10.1016/j.xphs.2021.01.026
  • Peer Reviewed
• [Journal Article] Complementary Leucine Zippering System for Effective Intracellular Delivery of Proteins by Cell-Penetrating Peptides (2021)
  • Author(s): Mizuki Kitamatsu, Hiroki Yuasa, Takashi Ohtsuki, Hiroyuki Michiue
  • Journal Title: Bioorganic & Medicinal Chemistry Volume: 33 Pages: 116036-116041
  • DOI: 10.1016/j.bmc.2021.116036
  • Peer Reviewed
• [Journal Article] Photoinduced endosomal escape mechanism: a view from photochemical internalization mediated by CPP-photosensitizer conjugates. (2021)
  • Author(s): Soe, T. H., Watanabe, K., and Ohtsuki, T.
  • Journal Title: Molecules Volume: 26 Issue: 1 Pages: 36-36
  • DOI: 10.3390/molecules26010036
  • NAID: 120007004792
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Cell cycle dependence of apoptosis photo-triggered using peptide-photosensitizer conjugate. (2020)
  • Author(s): Kim, H., Watanabe, S., Kitamatsu, M., Watanabe, K., and Ohtsuki, T.
  • Journal Title: Scientific Reports Volume: 10 Issue: 1 Pages: 19087-19087
  • DOI: 10.1038/s41598-020-76100-7
  • NAID: 120006937496
  • Peer Reviewed / Open Access
• [Journal Article] Intracellular delivery of a peptide nucleic acid-based hybrid of an autophagy inducing peptide with a cell-penetrating peptide (2020)
  • Author(s): Hakata Yoshiyuki、Ishikawa Suzuka、Ohtsuki Takashi、Miyazawa Masaaki、Kitamatsu Mizuki
  • Journal Title: Organic & Biomolecular Chemistry Volume: 18 Issue: 10 Pages: 1978-1986
  • DOI: 10.1039/c9ob02559f
  • Peer Reviewed
• [Journal Article] Endosomal escape of peptide-photosensitizer conjugates is affected by amino acid sequences near the photosensitizer. (2020)
  • Author(s): Miyoshi, Y., Kadono, M., Okazaki, S., Nishimura, A., Kitamatsu, M., Watanabe, K., and Ohtsuki, T.
  • Journal Title: Bioconjugate Chemistry Volume: 31 Issue: 3 Pages: 916-922
  • DOI: 10.1021/acs.bioconjchem.0c00046
  • Peer Reviewed
• [Journal Article] Combined apoptotic effects of peptide and miRNA in a peptide/miRNA nanocomplex. (2019)
  • Author(s): Kim, H., Kitamatsu, M., Ohtsuki, T.
  • Journal Title: Journal of Bioscience and Bioengineering Volume: 128 Issue: 1 Pages: 110-116
  • DOI: 10.1016/j.jbiosc.2019.01.003
  • NAID: 40021954126
  • Peer Reviewed
• [Journal Article] Relation of photochemical internalization to heat, pH, and Ca2+ ions. (2019)
  • Author(s): Soe, T. H., Nanjo, T., Watanabe, K., and Ohtsuki, T.
  • Journal Title: Photochemistry and Photobiology Volume: 95 Issue: 6 Pages: 1395-1402
  • DOI: 10.1111/php.13146
  • Peer Reviewed
• [Journal Article] Red and Near-Infrared Light-Directed Cytosolic Delivery of Two Different RNAs Using Photosensitive RNA Carriers (2018)
  • Author(s): Shiraga Kaori、Soe Tet Htut、Matsumoto Sho、Watanabe Kazunori、Ohtsuki Takashi
  • Journal Title: Bioconjugate Chemistry Volume: 29 Issue: 9 Pages: 3174-3179
  • DOI: 10.1021/acs.bioconjchem.8b00487
  • Peer Reviewed
• [Journal Article] 創薬を支える光技術 (2018)
  • Author(s): 渡邉 和則、大槻 高史
  • Journal Title: 月刊光アライアンス（6月号） Volume: - Pages: 1-4
• [Presentation] PCDR法の副作用低減 (2022)
  • Author(s): 坂東晃成、渡邉和則、大槻高史
  • Organizer: RNAフロンティアミーティング
• [Presentation] Cell cycle dependence of apoptosis photo-triggered using a peptide-photosensitizer conjugate. (2021)
  • Author(s): Takashi Ohtsuki, Hyungjin Kim, Mizuki Kitamatsu, Kazunori Watanabe
  • Organizer: Pacifichem 2021
  • Int'l Joint Research
• [Presentation] 細胞内へのタンパク質輸送のためPCI分子素子の開発 (2021)
  • Author(s): 角菜々子、北松瑞生、阿部由佳、渡邉和則、大槻高史
  • Organizer: 第94回日本生化学会大会
• [Presentation] 細胞内還元環境下でCPPが離脱するCPP融合タンパク質の開発 (2021)
  • Author(s): 小林達哉、中山真伍渡邉和則、大槻高史
  • Organizer: 日本化学会中国四国支部大会
• [Presentation] 光で細胞機能を操るための光応答分子の開発 (2020)
  • Author(s): 大槻高史
  • Organizer: 第24回関西大学先端科学技術シンポジウム
  • Invited
• [Presentation] 光で細胞機能を操るための光応答分子の開発 (2020)
  • Author(s): 大槻高史
  • Organizer: 第13回徳島文理大学 薬学部学術講演会
  • Invited
• [Presentation] Design of CPP-cargo-photosensitizer conjugates for effective photochemical internalization (2019)
  • Author(s): Miyoshi, Y., Kadono, M., Kitamatsu, M., Watanabe, K., Ohtsuki, T.
  • Organizer: 20th Tetrahedron Symposium
  • Int'l Joint Research
• [Presentation] Red and near-infrared light-directed cytosolic delivery of two different RNAs using photosensitive RNA carriers (2019)
  • Author(s): Ohtsuki, T., Shiraga, S., Soe, T.H, Matsumoto, S., Watanabe, K.
  • Organizer: 20th Tetrahedron Symposium
  • Int'l Joint Research
• [Presentation] 光応答的に働く卵活性化因子の開発 (2019)
  • Author(s): 大本 和正、若井 拓哉、舟橋 弘晃、渡邉 和則、大槻 高史
  • Organizer: 第42回 日本分子生物学会
• [Presentation] 細胞内還元環境下でCPPが離脱するCPP融合タンパク質の開発 (2019)
  • Author(s): 中山真伍、渡邊和則、大槻高史
  • Organizer: 日本化学会中国四国支部大会
• [Presentation] 光依存的に細胞質内に侵入する機能性ペプチドの設計 (2019)
  • Author(s): 三好祐一、北松瑞生、渡邉和則、大槻高史
  • Organizer: 第13回バイオ関連化学シンポジウム2019
• [Presentation] Relation of photochemical internalization to heat, pH, and Ca2+ ions (2019)
  • Author(s): Tet Htut Soe, Tomotaka Nanjo, Kazunori Watanabe, Takashi Ohtsuki
  • Organizer: 第13回バイオ関連化学シンポジウム2019
• [Presentation] 光依存的卵活性化因子の開発 (2018)
  • Author(s): 大本和正・若井拓哉・舟橋弘晃・渡邉和則・大槻高史
  • Organizer: 日本化学会中国四国支部大会
• [Presentation] pre-miR-664aによる光依存的なアポトーシス誘導法の開発 (2018)
  • Author(s): 縄稚 朋子、渡邉 和則、大槻 高史
  • Organizer: 第41回日本分子生物学会年会
• [Presentation] 光依存的なRNA導入法による２種類のRNAの時間差導入 (2018)
  • Author(s): 大槻高史, 白神 かおり, 渡邉 和則
  • Organizer: 第91回日本生化学会大会
• [Presentation] 光に応答し細胞質内に侵入する機能性光増感剤ペプチドの設計 (2018)
  • Author(s): 三好 祐一, 山本 怜見, 門野 真保, 北松 瑞生, 渡邉 和則, 大槻 高史
  • Organizer: 第91回日本生化学会大会
• [Presentation] 光応答性キャリアを用いたphotochemical internalizationの機構解明 (2018)
  • Author(s): 南條 友孝, 渡邉 和則, 大槻 高史
  • Organizer: 第91回日本生化学会大会
• [Presentation] PCDR法による細胞質内RNA送達の原理と応用 (2018)
  • Author(s): 大槻高史、Soe Tet Htut、渡邉 和則
  • Organizer: 第56回 日本生物物理学会年会
• [Book] 核酸科学ハンドブック (2020)
  • Author(s): 渡邉 和則、大槻 高史（[RNAデリバリー]の項を分担執筆）
  • Total Pages: 576
  • Publisher: 講談社サイエンティフク
  • ISBN: 9784065207864