Development of lipid nanoparicles for treatment of hepatic diseases by in vivo genome editing

• Project/Area Number: 17H05052
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Single-year Grants
• Research Field: Physical pharmacy
• Research Institution: Hokkaido University
• Principal Investigator: Sato Yusuke 北海道大学, 薬学研究院, 助教 (10735624)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥22,880,000 (Direct Cost: ¥17,600,000、Indirect Cost: ¥5,280,000); Fiscal Year 2019: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2018: ¥6,110,000 (Direct Cost: ¥4,700,000、Indirect Cost: ¥1,410,000); Fiscal Year 2017: ¥11,700,000 (Direct Cost: ¥9,000,000、Indirect Cost: ¥2,700,000)
• Keywords: ゲノム編集 / 脂質ナノ粒子 / マイクロ流体デバイス / in vivoゲノム編集 / RNP送達 / pH感受性カチオン性脂質 / 肝疾患治療 / ph感受性カチオン性脂質 / 安全治療域 / タンパク質送達 / RNP / In vivoゲノム編集 / RNA送達

Outline of Final Research Achievements

Genome editing is a biotechnology that can disrupt, repair and insert genes of interest and can be applied for fundamental treatment of various hereditary diseases. Development of nanotechnology capable of efficient delivery of genome editing factors is important for medical application. In this study, lipid nanoparticles that deliver CRISPR-Cas protein-gRNA complexes efficiently were developed. From a viewpoint of practical application, microfluidic device-based synthesizing methodology that enables seamless manufacturing scale-up was developed.

Academic Significance and Societal Importance of the Research Achievements

マイクロ流体デバイスによるゲノム編集因子搭載脂質ナノ粒子の連続製造法を世界に先駆けて見出すことに成功した。マイクロ流体デバイスはシームレスな製造スケールアップが可能であることから、核酸ナノ医薬品開発に広く応用されているが、核酸分子と比較して物理化学的に不安定なタンパク質のナノ製剤化へ応用できることを示したことは今後の高分子搭載ナノ粒子製剤開発における意義は大きい。また、製剤最適化により競合技術よりも圧倒的に高い効率で遺伝子ノックアウトを誘導可能な脂質ナノ粒子製剤の同定に成功した。最適化の過程で得られた重要因子に関する情報は今後の製剤開発における指針となると考えられる。

Research Products

• [Journal Article] Hydrophobic scaffolds of pH-sensitive cationic lipids contribute to miscibility with phospholipids and improve the efficiency of delivering short interfering RNA by small-sized lipid nanoparticles (2020)
  • Author(s): Sato Y, Okabe N, Note Y, Hashiba K, Maeki M, Tokeshi M, Harashima H
  • Journal Title: Acta Biomaterialia Volume: 102 Pages: 341-350
  • DOI: 10.1016/j.actbio.2019.11.022
  • Peer Reviewed
• [Journal Article] Innovative nanotechnologies for enhancing nucleic acids/gene therapy: Controlling intracellular trafficking to targeted biodistribution (2019)
  • Author(s): Nakamura Takashi, Yamada Yuma, Sato Yusuke, Khalil A Ikramy, Harashima Hideyoshi
  • Journal Title: Biomaterials Volume: 218 Pages: 119329-119329
  • DOI: 10.1016/j.biomaterials.2019.119329
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Recent advances in the targeting of systemically administered non-viral gene delivery systems. (2019)
  • Author(s): Khalil IA, Sato Y, Harashima H.
  • Journal Title: Expert Opin Drug Deliv. Volume: 16 Issue: 10 Pages: 1037-1050
  • DOI: 10.1080/17425247.2019.1656196
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Understanding structure-activity relationships of pH-sensitive cationic lipids facilitates the rational identification of promising lipid nanoparticles for delivering siRNAs in vivo (2019)
  • Author(s): Yusuke Sato, Kazuki Hashiba, Kosuke Sasaki, Masatoshi Maeki, Manabu Tokeshi, HIdeyoshi Harashima
  • Journal Title: Journal of Controlled Release Volume: 295 Pages: 140-152
  • DOI: 10.1016/j.jconrel.2019.01.001
  • Peer Reviewed
• [Journal Article] Development of the iLiNP Device: Fine Tuning the Lipid Nanoparticle Size within 10 nm for Drug Delivery (2018)
  • Author(s): Niko Kimura, Masatoshi Maeki, Yusuke Sato, Yusuke Note, Akihiko Ishida, Hirofumi Tani, Hideyoshi Harashima, Manabu Tokeshi
  • Journal Title: ACS Omega Volume: 3 Issue: 5 Pages: 5044-5041
  • DOI: 10.1021/acsomega.8b00341
  • Peer Reviewed / Open Access
• [Journal Article] Understanding the formation mechanism of lipid nanoparticles in microfluidic devices with chaotic micromixers (2017)
  • Author(s): Maeki M, Fujishima Y, Sato Y, Yasui T, Kaji N, Ishida A, Tani H, Baba Y, Harashima H, Tokeshi M
  • Journal Title: PLoS One Volume: 12 Issue: 11 Pages: e0187962-e0187962
  • DOI: 10.1371/journal.pone.0187962
  • NAID: 120006377541
• [Journal Article] Sato Y, Matsui H, Yamamoto N, Sato R, Munakata T, Kohara M, Harashima H. Highly specific delivery of siRNA to hepatocytes circumvents endothelial cell-mediated lipid nanoparticle-associated toxicity leading to the safe and efficacious decrease in the hepatitis B virus (2017)
  • Author(s): Sato Y, Matsui H, Yamamoto N, Sato R, Munakata T, Kohara M, Harashima H
  • Journal Title: J Control Release Volume: 266 Pages: 216-225
  • DOI: 10.1016/j.jconrel.2017.09.044
  • Peer Reviewed
• [Journal Article] pH-labile PEGylation of siRNA-loaded lipid nanoparticle improves active targeting and gene silencing activity in hepatocytes (2017)
  • Author(s): Hashiba K, Sato Y, Harashima H
  • Journal Title: J. Control. Release Volume: 262 Pages: 239-246
  • DOI: 10.1016/j.jconrel.2017.07.046
  • NAID: 120006523729
  • Peer Reviewed
• [Presentation] Efficient delivery of macromolecules by molecular design of cationic lipids (2019)
  • Author(s): Sato Y, Hashiba K, Hashiba A, Okabe N, Harashima H
  • Organizer: Liposome Research Days 2019
  • Int'l Joint Research
• [Presentation] Development of Ribonucleoprotein-loaded lipid nanoparticles for genome editing (2019)
  • Author(s): Suzuki Y, Sato Y, Sato R, Maeki M, Tokeshi M, Harashima H
  • Organizer: Liposome Research Days 2019
  • Int'l Joint Research
• [Presentation] ゲノム編集タンパク質搭載脂質ナノ粒子の開発 (2019)
  • Author(s): 佐藤悠介, 鈴木裕一, 佐藤里咲, 真栄城正寿, 渡慶次学, 原島秀吉
  • Organizer: 日本核酸医薬学学会第5回年会
• [Presentation] Cas9-gRNA RNP搭載脂質ナノ粒子による高効率ゲノム編集 (2019)
  • Author(s): 佐藤悠介, 鈴木裕一, 佐藤里咲, 真栄城正寿, 渡慶次学, 原島秀吉
  • Organizer: 日本ゲノム編集学会第4回大会
• [Presentation] 優れたsiRNA送達効率と安全性を両立する脂質ナノ粒子の開発 (2018)
  • Author(s): 佐藤悠介, 橋場一毅, 原島秀吉.
  • Organizer: 日本薬学会第137年会
• [Presentation] 効率的なin vivo核酸送達を実現する高機能脂質ナノ粒子 (2018)
  • Author(s): 佐藤悠介, 橋場一毅, 原島秀吉.
  • Organizer: 第27回インテリジェント材料・システムシンポジウム
• [Presentation] Targeted delivery of siRNAs by manipulating acid dissociation constant of lipid nanoparticles (2017)
  • Author(s): Shobaki N, Sato Y, Harashima H.
  • Organizer: The 5th International Life Science Symposium for Young Scientists
  • Int'l Joint Research
• [Presentation] 機能性脂質を基盤としたin vivo核酸送達システムの開発 (2017)
  • Author(s): 佐藤悠介.
  • Organizer: 日本薬学会北海道支部主催 第1回若手シンポジウム
  • Invited
• [Presentation] 新規pH応答性脂質の開発による効率的なin vivo siRNAデリバリー (2017)
  • Author(s): 橋場一毅, 佐藤悠介, 原島秀吉.
  • Organizer: 第39回生体膜と薬物の相互作用シンポジウム
• [Presentation] 脂質ナノ粒子の膜酸乖離定数の操作による細胞選択的なsiRNA送達 (2017)
  • Author(s): ショバキ ヌレ, 佐藤悠介, 原島秀吉.
  • Organizer: 第3回日本核酸医薬学会
• [Patent(Industrial Property Rights)] 脂質ナノ粒子 (2019)
  • Inventor(s): 佐藤悠介, 原島秀吉, 渡慶次学, 真栄城正寿
  • Industrial Property Rights Holder: 国立大学法人北海道大学
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2019-101203
  • Filing Date: 2019
• [Patent(Industrial Property Rights)] siRNA細胞内送達のための脂質膜構造体 (2018)
  • Inventor(s): 原島秀吉, 佐藤悠介
  • Industrial Property Rights Holder: 国立大学法人北海道大学
  • Industrial Property Rights Type: 特許
  • Filing Date: 2018
  • Overseas
• [Patent(Industrial Property Rights)] siRNA細胞内送達のための脂質膜構造体 (2017)
  • Inventor(s): 原島秀吉, 佐藤悠介
  • Industrial Property Rights Holder: 国立大学法人北海道大学
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2017-117708
  • Filing Date: 2017