Realization of effective gene transfection by actively DNA folding-controllable polyplexes

• Project/Area Number: 17H02102
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biomedical engineering/Biomaterial science and engineering
• Research Institution: Osaka Prefecture University
• Principal Investigator: Harada Atsushi 大阪府立大学, 工学(系)研究科(研究院), 教授 (50302774)
• Project Period (FY): 2017-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000); Fiscal Year 2020: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000); Fiscal Year 2019: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000); Fiscal Year 2018: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2017: ¥7,410,000 (Direct Cost: ¥5,700,000、Indirect Cost: ¥1,710,000)
• Keywords: 遺伝子ベクター / DNAフォールディング / アスペクト比 / 転写過程 / 多分岐ポリマー / カチオン性ポリマー / 温度変化 / ポリカチオン / 折りたたみ制御

Outline of Final Research Achievements

In this study, we synthesized a cationic polymer with a multi-branched polymer structure in order to actively control the property that charge-neutralized plasmid DNA is regularly folded, and then, we tried to control molecular folding of DNA in the polyplex. By focusing on the density in the polyplex of the multi-branched polymer part, various DNA molecule folding states were induced, and the correlation between the folding state and the transcription efficiency was clarified. Polyplexes with optimized DNA molecule folding status exhibited highly efficient gene transfection.

Academic Significance and Societal Importance of the Research Achievements

がんなどの難治性の疾患の遺伝子治療やワクチンなどの免疫治療において、合成材料を用いて高効率な遺伝子発現を可能とするベクター開発は重要な課題となっています。本研究での研究成果は、遺伝子を発現させたい細胞に到達するまでは安定に遺伝子（DNA）を保持し、細胞へ到達した後にDNAを働かせやすい状態へと変化させることによって遺伝子発現効率の向上が可能であることを示しました。DNAだけでなくmRNAなどへの適応性を検討することでさまざまな疾患治療を可能とさせると期待されます。

Research Products

• [Journal Article] Comparison of In Vitro Performances of Nanorod and Nanofiber Polyplexes Prepared from Plasmid DNA and Poly(L-lysine) Terminally Bearing Multi-Arm PEG (2020)
  • Author(s): Ryuta Aono, Kenta Nomura, Eiji Yuba, Atsushi Harada
  • Journal Title: Polymers in Therapeutic Delivery, ACS Symposium Series Volume: 1350 Pages: 13-21
  • DOI: 10.1021/bk-2020-1350.ch002
  • ISBN: 9780841238145, 9780841298996
  • Peer Reviewed
• [Journal Article] Preparation of photothermal-chemotherapy nanohybrids by complexation of gold nanorods with polyamidoamine dendrimers having poly(ethylene glycol) and hydrophobic chains (2020)
  • Author(s): Hashimoto Takuya、Yuba Eiji、Harada Atsushi、Kono Kenji
  • Journal Title: Journal of Materials Chemistry B Volume: 8 Issue: 14 Pages: 2826-2833
  • DOI: 10.1039/c9tb02163a
  • NAID: 120006938354
• [Journal Article] Light-Activatable Transfection System Using Hybrid Vectors Composed of Thermosensitive Dendron Lipids and Gold Nanorods (2020)
  • Author(s): Hashimoto Takuya、Hirata Tomoya、Yuba Eiji、Harada Atsushi、Kono Kenji
  • Journal Title: Pharmaceutics Volume: 12 Issue: 3 Pages: 239-239
  • DOI: 10.3390/pharmaceutics12030239
• [Journal Article] Temperature-Responsive Molecular Assemblies Using Oligo(Ethylene Glycol)-Attached Polyamidoamine Dendron Lipids and their Functions as Drug Carriers (2020)
  • Author(s): Hashimoto Takuya、Hirai Yuji、Yuba Eiji、Harada Atsushi、Kono Kenji
  • Journal Title: Journal of Functional Biomaterials Volume: 11 Issue: 1 Pages: 16-16
  • DOI: 10.3390/jfb11010016
• [Journal Article] Gene Expression of Aspect Ratio-Controlled Polyplexes Based on the Effect of Multi-Arm Poly(ethylene glycol) (2019)
  • Author(s): Harada Atsushi、Nomura Kenta、Yuba Eiji、Kono Kenji
  • Journal Title: ACS Biomaterials Science & Engineering Volume: in press Issue: 11 Pages: 5681-5687
  • DOI: 10.1021/acsbiomaterials.8b01498
  • Peer Reviewed
• [Presentation] Near infrared light-activated transfection system using hybrid vectors composed of thermosensitive dendron lipids and gold nanorods (2019)
  • Author(s): T. Hashimoto, E. Yuba, A. Harada, K. Kono
  • Organizer: 19th Symposium for Gene Design and Delivery
  • Int'l Joint Research
• [Presentation] 多分岐PEG導入ポリリシンからなるナノファイバーポリプレックスへの温度応答性付与 (2019)
  • Author(s): 原田 敦史，勝圓 由紀子，弓場 英司
  • Organizer: 第48回医用高分子シンポジウム
• [Presentation] Gene Expression of Aspect-Ratio-Controlled Polyplexes Based on the Effect of Multi-Arm Poly(ethylene glycol) (2019)
  • Author(s): Atsushi Harada, Eiji Yuba
  • Organizer: ACS Spring 2019 National Meeting
  • Int'l Joint Research
• [Presentation] PEG鎖連結高分子電解質の自己組織化におけるPEG鎖の役割 (2018)
  • Author(s): 原田敦史
  • Organizer: 第36回関西界面科学セミナー：ソフトコロイドの界面構造および機能発現－基礎から応用まで
  • Invited
• [Presentation] 高分子電解質へのPEG鎖導入効果 (2018)
  • Author(s): 原田敦史
  • Organizer: 第38回キャピラリー電気泳動シンポジウム
  • Invited
• [Presentation] Aspect ratio-controllable polyplexes prepared from poly(L-lysine) terminally bearing multi-arm PEG (2018)
  • Author(s): A. Harada, E. Yuba, K. Kono
  • Organizer: 255th ACS National Meeting & Expositions
  • Int'l Joint Research
• [Presentation] siRNAデリバリーのための高分子材料設計 (2017)
  • Author(s): 原田 敦史，山本 聡，大村 啓輔，弓場 英司
  • Organizer: 日本核酸医薬学会 第3回年会
  • Invited
• [Presentation] 多分岐PEG被覆ポリプレックスの形態及び表面特性制御 (2017)
  • Author(s): 原田 敦史，勝圓 由紀子，野村 健太，弓場 英司，河野 健司
  • Organizer: 第66回高分子討論会