Establishment of the third generation plasma gene introduction method by suppressing autophagy and by enhancing synergistic effect

• Project/Area Number: 17H01068
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Plasma electronics
• Research Institution: Ehime University
• Principal Investigator: JINNO MASAFUMI 愛媛大学, 理工学研究科(工学系), 教授 (30274335)
• Project Period (FY): 2017-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥41,990,000 (Direct Cost: ¥32,300,000、Indirect Cost: ¥9,690,000); Fiscal Year 2020: ¥8,840,000 (Direct Cost: ¥6,800,000、Indirect Cost: ¥2,040,000); Fiscal Year 2019: ¥8,840,000 (Direct Cost: ¥6,800,000、Indirect Cost: ¥2,040,000); Fiscal Year 2018: ¥12,090,000 (Direct Cost: ¥9,300,000、Indirect Cost: ¥2,790,000); Fiscal Year 2017: ¥12,220,000 (Direct Cost: ¥9,400,000、Indirect Cost: ¥2,820,000)
• Keywords: プラズマ遺伝子導入 / プラズマ分子導入 / プラズマライフサイエンス / エンドサイトーシス / 複合効果 / 低侵襲 / 非侵襲 / ランダムインテグレーション / オートファジー / プラズマ医療 / 遺伝子導入 / シナジー効果 / 高効率 / 複合刺激 / プラズマ / 遺伝子 / 形質転換

Outline of Final Research Achievements

The results suggest that plasma does not only induce endocytosis but also promotes membrane attachment of plasmids since the mere induction of endocytosis by endocytosis-promoting agents without plasma stimulation hardly results in gene transfer. The results of the circuit analysis using an equivalent network model of the cell population in the well suggested that the electric current is the main factor of electrical stimulation.
We conclude that plasma gene transfer occurs in a two-step process: the acceleration of the plasmid-cell collision process by cell charging and the induction of endocytosis by the combined stimulation of electric current and reactive oxygen species.

Academic Significance and Societal Importance of the Research Achievements

プラズマ遺伝子・分子導入法の機序解明が進み、プラズマの必要性とエンドサイトーシスでの分子取り込みに必要なプラズマの作用要因が明確になった。特に、電気的要因として電流が主要であることが示唆されたことは意義が大きい。
また、社会的には、従来法とは異なり、細胞へのダメージが小さくかつ効率良く導入を行えるプラズマ法の機序解明が進んだことで、実用へ大きく近づくことが出来た。

Research Products

• [Journal Article] Importance of collision frequency in the molecular size dependency of gene transfer efficiency in surface discharge method (2019)
  • Author(s): Tatsuya Hiramatsu, Hiroko Hirashige, Yugo Kido, Susumu Satoh, and Masafumi Jinno
  • Journal Title: Japanese Journal of Applied Physics Volume: 1 Pages: 1-1
  • NAID: 210000156277
  • Peer Reviewed
• [Presentation] Gene transfer efficiency of surface discharge method depending on molecular size and collision frequency between gene and cell (2019)
  • Author(s): M. Jinno, T. Hiramatsu, H. Hirashige, Y. Kido, and S. Satoh
  • Organizer: 24th International Symposium on Plasma Chemistry
  • Int'l Joint Research
• [Presentation] Informatics optimization of the parameters for individual cell kind in micro-plasma gene transfection (2019)
  • Author(s): Jinno, M., Yoshitake, T., Miyamoto, N., Kido, Y. and Satoh, S.
  • Organizer: 2nd International Conference on Data Driven Plasma Science
  • Int'l Joint Research
• [Presentation] Informatics Approach Considering Tree Dimensional Cell Structure to Optimize Plasma Irradiation Time in Plasma Gene Transfer Method (2019)
  • Author(s): Yoshitake, T., Miyamoto, N., Kido, Y., Satoh, S., and Jinno, M
  • Organizer: 2nd International Conference on Data Driven Plasma Science
  • Int'l Joint Research
• [Presentation] Innovative Gene/Molecule Transfection Using Micro-plasma (2018)
  • Author(s): Masafumi Jinno, Yoshihisa Ikeda, Yugo Kido, and Susumu Satoh
  • Organizer: 10th anniversary of International Workshop on Microplasmas
  • Int'l Joint Research
• [Presentation] Improve gene stability in plasma gene transfection by autophagy inhibition (2018)
  • Author(s): Yuto Kikuchi, Yuki Isozaki, Yoshihisa Ikeda, Yugo Kido, Susumu Satoh, Masafumi Jinno
  • Organizer: 7th International Conference on Plasma Medicine
  • Int'l Joint Research
• [Presentation] Endocytosis dependency of gene transfer in plasma method and electroporation (2018)
  • Author(s): Masafumi Jinno, Kouki Yamamoto, Yugo Kido, Susumu Satoh
  • Organizer: 7th International Conference on Plasma Medicine
  • Int'l Joint Research
• [Presentation] プラズマ遺伝子導入法における細胞種毎の最適条件へのインフォマティックス的アプローチ (2018)
  • Author(s): 吉武 卓哉,宮本 展寛,木戸 祐吾,佐藤 晋,神野 雅文
  • Organizer: 第66回応用物理学会春季学術講演会
• [Presentation] プラズマ遺伝子導入におけるプラズマ照射条件のインフォマティックス的最適化 (2018)
  • Author(s): 吉武 卓哉,宮本 展寛,木戸 祐吾,佐藤 晋,神野 雅文
  • Organizer: 第79回応用物理学会秋季学術講演会
• [Presentation] プラズマ照射による上皮細胞への遺伝子導入 (2018)
  • Author(s): 吉武 卓哉、宮本 展寛、木戸 祐吾、佐藤 晋、神野 雅文
  • Organizer: 2018年第65回応用物理学会春季学術講演会
• [Presentation] 塩化マンガンが遺伝子発現に及ぼす効果 (2018)
  • Author(s): 森 貴之、木戸 祐吾、池田 善久、佐藤 晋、神野 雅文
  • Organizer: 2018年第65回応用物理学会春季学術講演会
• [Presentation] Endocytosis Dependency of Gene Transfer in Plasma Method and Electroporation (2018)
  • Author(s): M. Jinno、K. Yamamoto、Y. Kido、S. Satoh
  • Organizer: The 7th International Conference on Plasma Medicine
  • Int'l Joint Research
• [Presentation] Improve gene stability in plasma gene transfection by autophagy inhabitation (2018)
  • Author(s): Y. Kikuchi、Y. Isozaki、Y. Ikeda、Y. Kido、S. Satoh、M. Jinno
  • Organizer: The 7th International Conference on Plasma Medicine
  • Int'l Joint Research
• [Presentation] プラズマ遺伝子導入における電気的要因と化学的要因のシナジー効果 (2017)
  • Author(s): 池田 善久、木戸 祐吾、佐藤 晋、神野 雅文
  • Organizer: 2017年第78回応用物理学会秋季学術講演会
• [Presentation] プラズマ遺伝子・分子導入における電気的作用要因の寄与 (2017)
  • Author(s): 木戸 祐吾、本村 英樹、佐藤 晋、神野 雅文
  • Organizer: 2017年第78回応用物理学会秋季学術講演会
• [Patent(Industrial Property Rights)] 形質転換細胞の製造方法 (2018)
  • Inventor(s): 神野雅文、加藤英政、徳澤佳美、大西章仁、明上純子、佐藤晋
  • Industrial Property Rights Holder: 神野雅文、加藤英政、徳澤佳美、大西章仁、明上純子、佐藤晋
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2018-115200
  • Filing Date: 2018