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2018 年度 実施状況報告書

Fine-tuning of polymer amphiphile for high performance of messenger RNA delivery

研究課題

研究課題/領域番号 18K18378
研究機関東京大学

研究代表者

Kim HyunJin  東京大学, 大学院医学系研究科(医学部), 特任助教 (10755002)

研究期間 (年度) 2018-04-01 – 2020-03-31
キーワードcationic polymer / mRNA transfection / amphiphile
研究実績の概要

Precursor polymers was reacted with endosomal escape moiety group (diethylenetriamine, DET) and various aliphatic amine groups (pentyl, heptyl, octyl, nonyl, decyl, 2-cyclohexylethyl (CHE)), 2-phenylethyl amines) to make amphiphilic polymers. All aliphatic amine groups were successfully introduced to the polymer at around 40% of total degree of polymerization. A series of amphiphilic polymers formed 100-150 nm sized nanoparticles with firefly or gaussian luciferase mRNA. These nanoparticles were transfected into cultured mouse myoblast cells and neuronal cancer cells. Amphiphilic polymers installed with DET and CHE moieties (CHE-polymer) showed the highest translation efficacy and exhibited 5 orders of magnitude higher than the other polymer with the least efficacy. The CHE-polymer also showed better Cas9 mRNA translation efficacy in luciferase-based gene editing assay, compared to commercially available lipofectamine. We confirmed that there is optimal hydrophobicity for efficient mRNA transfection in cultured cells, i.g. between particle stability and release of mRNA in cytoplasm. Thus, fine-tuning of hydrophobicity in the amphiphilic polymer dramatically increases in vitro transfection efficacy of mRNA.

現在までの達成度 (区分)
現在までの達成度 (区分)

2: おおむね順調に進展している

理由

CHE-polymer showed the extremely higher mRNA transfection efficacy in cultured cell condition than initially expected. Especially CHE-polymer showed higher Cas9 mRNA transfection efficacy in luciferase-based gene editing assay than lipofectamine. This is the great improvement of cationic polymer development.

今後の研究の推進方策

1. In fiscal year 2019, a series of amphiphilic polymer will be investigated for delivery efficacy of mRNA in mouse animals. Firefly luciferase mRNA-loaded particles will be administered into mouse in the various injection manners and their luminescence intensities will be measured by in vivo imaging system. These experiments will provides the clues that the hydrophobicity tuning will also work in vivo condition.
2. The future work tries to find out the next parameter for the second round polymer synthesis. If possible, the second round amphiphilic polymers will be synthesized based on the new design parameter.

次年度使用額が生じた理由

I successfully found amphiphilic polymer candidate with high performance for mRNA delivery earlier than I expected. I already finished particle characterization and mRNA transfection efficacy in cultured cells. Now, I want to investigate mRNA delivery efficacy of the amphiphilic polymer in mouse animal. High doses of polymer and mRNA amounts are necessary for animal experiments. In this reason, I want to transfer research fund in fiscal year 2018 into fiscal year 2019.

  • 研究成果

    (6件)

すべて 2019 2018

すべて 雑誌論文 (1件) (うち査読あり 1件) 学会発表 (4件) (うち国際学会 2件) 産業財産権 (1件)

  • [雑誌論文] Glucose-linked sub-50-nm unimer polyion complex-assembled gold nanoparticles for targeted siRNA delivery to glucose transporter 1-overexpressing breast cancer stem-like cells2019

    • 著者名/発表者名
      Yi Yu、Kim Hyun Jin、Zheng Meng、Mi Peng、Naito Mitsuru、Kim Beob Soo、Min Hyun Su、Hayashi Kotaro、Perche Federico、Toh Kazuko、Liu Xueying、Mochida Yuki、Kinoh Hiroaki、Cabral Horacio、Miyata Kanjiro、Kataoka Kazunori
    • 雑誌名

      Journal of Controlled Release

      巻: 295 ページ: 268~277

    • DOI

      10.1016/j.jconrel.2019.01.006

    • 査読あり
  • [学会発表] Tuning of Hydrophobicity in Amphiphilic Polymers for mRNA Delivery and Their Application for CRISPR-Cas9 Technology2019

    • 著者名/発表者名
      Hyun Jin Kim, Kazunori Kataoka, Kanjiro Miyata
    • 学会等名
      12th Annual Symposium on Nanobiotechnology
    • 国際学会
  • [学会発表] Adjustment of Hydrophobicity in Amphiphilic Polymers For Highly Efficient Messenger RNA Delivery2019

    • 著者名/発表者名
      Hyun Jin Kim, Moritoshi Sato, Kazunori Kataoka, Kanjiro Miyata
    • 学会等名
      6th International Conference on Multifunctional, Hybrid and Nanomaterials
    • 国際学会
  • [学会発表] Development of amphiphilic polymers for CRISPR-Cas9 mRNA delivery2018

    • 著者名/発表者名
      Hyun Jin Kim, Kazunori Kataoka, Kanjiro Miyata
    • 学会等名
      5th COINS Symposium
  • [学会発表] Development of Thiol-Crosslinked Micelle for Systemic Delivery of Antisense Oligonucleotide to Treat Brain Disease2018

    • 著者名/発表者名
      Hyun Jin Kim, Hyun Su Min, Keisaku Katsushima, Yutaka Kondo, Kanjiro Miyata, Kazunori Kataoka
    • 学会等名
      第67回高分子学会年次大会
  • [産業財産権] 両親媒性ポリアミノ酸、該両親媒性ポリアミノ酸を用いたブロックコポリマー、および該両親媒性ポリアミノ酸または該ブロックコポリマーと核酸とを含む複合体2018

    • 発明者名
      キムヒョンジュン, 宮田 完二郎、片岡一則
    • 権利者名
      キムヒョンジュン, 宮田 完二郎、片岡一則
    • 産業財産権種類
      特許
    • 産業財産権番号
      2018-178320

URL: 

公開日: 2019-12-27  

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