Development of Gene Delivery Method to Control of Autophagy mechanisms

• Project/Area Number: 20K07029
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 47030:Pharmaceutical hygiene and biochemistry-related
• Research Institution: Osaka University
• Principal Investigator: Ogawa Hidesato 大阪大学, 大学院生命機能研究科, 招へい教授 (20370132)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2022: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2021: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
• Keywords: 遺伝子導入 / 核酸医薬 / ウイルスベクター / オートファジー / アデノウィルス / 核酸導入 / ウイルス感染 / 化合物スクリーニング

Outline of Research at the Start

細胞は細胞外からの様々な分子の侵入を即座に検知し，排除するために選択的オートファジー機構を使用する。この機構は，外来異物だけでなく核酸の導入に対しても働くため，核酸導入が必要な場面で大きな障害となる。本研究では外来核酸のオートファジーによる分解に重要な役割を持つオートファジーレセプターp62に着目し，p62の機能を阻害し核酸導入効率を上昇させる低分子化合物のスクリーニングを行う。同定された化合物のp62に対する作用点を明らかにし，p62による核酸排除機構の解明を行うと共に，その効果をゲノム編集技術やマウス生体内への遺伝子導入において検討し，新規遺伝子導入促進試薬としての応用を検討する。

Outline of Final Research Achievements

Autophagy plays an important role in the defense against pathogen infection. Since most pathogens possess DNA or RNA, this system may play a role in the removal of foreign nucleic acids. The selective autophagy receptor p62 can rapidly degrade transfected nucleic acids and significantly inhibit gene delivery. High throughput screening of chemical compounds has shown that the inhibition of TBK1, which controls p62 activity, significantly increases gene transfection efficiency. In addition, these inhibitors can also enhance adenovirus infection. It was also found that adenoviral proteins form the complex that inhibits p62. These research results are expected to contribute not only to nucleic acid delivery, but also to medical applications using viral vectors.

Academic Significance and Societal Importance of the Research Achievements

本研究成果は、遺伝子導入やウイルス感染などの分野で大きな社会的な意義を持っている。 遺伝子導入においては、p62が細胞内で核酸の分解を促進することに着目し、p62の機能抑制による遺伝子導入効率の促進は、新たな遺伝子治療法の開発につながることが予測される。また世界中で問題となったCOVID-19パンデミックにおいて、ワクチンの開発は極めて重要な課題であり、そのなかで本研究は、アデノウイルス感染制御剤を可能とすることで危険性の少ないアデノウイルスベクターを用いたワクチンの基盤技術を提供することが期待される。

Research Products

• [Int'l Joint Research] Inst. Biotech of Polish Academy of Sci.(ポーランド)
• [Int'l Joint Research] Dept of Obste, Gynecology, and Reproduct(カナダ)
• [Journal Article] TBK1 inhibitors enhance transfection efficiency by suppressing p62/SQSTM1 phosphorylation (2023)
  • Author(s): Tsuchiya M, Kong W, Hiraoka Y, Haraguchi T, Ogawa H
  • Journal Title: Genes to Cells Volume: 28(1) Issue: 1 Pages: 68-77
  • DOI: 10.1111/gtc.12987
  • Peer Reviewed / Open Access
• [Journal Article] Transcriptional coregulator Ess2 controls survival of post-thymic CD4+ T cells through the Myc and IL-7 signaling pathways (2022)
  • Author(s): Takada Ichiro、Hidano Shinya、Takahashi Sayuri、Yanaka Kaori、Ogawa Hidesato、Tsuchiya Megumi、Yokoyama Atsushi、Sato Shingo、Ochi Hiroki、Nakagawa Tohru、Kobayashi Takashi、Nakagawa Shinichi、Makishima Makoto
  • Journal Title: Journal of Biological Chemistry Volume: 298 Issue: 9 Pages: 102342-102342
  • DOI: 10.1016/j.jbc.2022.102342
  • Peer Reviewed / Open Access
• [Journal Article] Age-associated decline of MondoA drives cellular senescence through impaired autophagy and mitochondrial homeostasis (2022)
  • Author(s): Yamamoto-Imoto Hitomi et al.,
  • Journal Title: Cell Reports Volume: 38 Issue: 9 Pages: 110444-110444
  • DOI: 10.1016/j.celrep.2022.110444
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Microtubule inhibitors identified through nonbiased screening enhance DNA transfection efficiency by delaying p62-dependent ubiquitin recruitment (2021)
  • Author(s): Megumi Tsuchiya, Hidesato Ogawa, Kento Watanabe, Takako Koujin, Chie Mori, Kazuto Nunomura, Bangzhong Lin, Akiyoshi Tani, Yasushi Hiraoka, Tokuko Haraguchi
  • Journal Title: Genes Cells. Volume: 26 Issue: 9 Pages: 739-751
  • DOI: 10.1111/gtc.12881
  • Peer Reviewed / Open Access
• [Journal Article] THOC4 regulates energy homeostasis by stabilizing TFEB mRNA during prolonged starvation (2021)
  • Author(s): Fujita T, Kubo S, Shioda T, Tokumura A, Minami S, Tsuchiya M, Ogawa H, Hamasaki M, Yu L, Isaka Y, Yoshimori T, and Nakamura S
  • Journal Title: Journal of Cell Science Volume: 134 Issue: 6 Pages: 248203-248203
  • DOI: 10.1242/jcs.248203
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] A missense mutation in the RSRSP stretch of Rbm20 causes dilated cardiomyopathy and atrial fibrillation in mice (2020)
  • Author(s): Ihara K, Sasano T, Hiraoka Y, Togo-Ohno M, Soejima Y, Sawabe M, Tsuchiya M, Ogawa H, Furukawa T, Kuroyanagi H
  • Journal Title: Scientific Reports Volume: 10 Issue: 1 Pages: 17894-17894
  • DOI: 10.1038/s41598-020-74800-8
  • NAID: 120007031666
  • Peer Reviewed / Open Access
• [Journal Article] Deletion of the Prdm3 Gene Causes a Neuronal Differentiation Deficiency in P19 Cells (2020)
  • Author(s): Leszczynski Pawel, Smiech Magdalena、Salam Teeli Aamir、Haque Effi、Viger Robert、Ogawa Hidesato、Pierzcha?a Mariusz、Taniguchi Hiroaki
  • Journal Title: International Journal of Molecular Sciences Volume: 21 Issue: 19 Pages: 7192-7192
  • DOI: 10.3390/ijms21197192
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] 選択的オートファジー制御を介した遺伝子導入促進剤のスクリーニング (2021)
  • Author(s): 小川 英知、土屋 惠、渡邊 賢人、荒神 尚子、森 知栄、布村 一人、林 邦忠、谷 昭義、平岡 泰、原口 徳子
  • Organizer: 第73回日本細胞生物学会大会
• [Presentation] High-Throughput Screening for the enhancement of transfection efficiency via selective autophagy (2020)
  • Author(s): Hidesato Ogawa, Megumi Tsuchiya, Kento Watanabe, Takako Koujin, Chie Mori, Kazuto Nunomura, Bangzhong Lin, Akiyoshi Tani, Yasushi Hiraoka, and Tokuko Haraguch
  • Organizer: 第43回日本分子生物学会年会
• [Remarks]
  • URL: https://www.fbs.osaka-u.ac.jp/labs/hiraoka/index.html
• [Remarks]
  • URL: https://www.fbs.osaka-u.ac.jp/labs/hiraoka/
• [Patent(Industrial Property Rights)] 2 種類の TBK1/IKKe 阻害剤を利用した核酸導入 (2021)
  • Inventor(s): 小川英知，土屋惠，渡邊賢人，平岡泰，原口徳子
  • Industrial Property Rights Holder: 小川英知，土屋惠，渡邊賢人，平岡泰，原口徳子
  • Industrial Property Rights Type: 特許
  • Filing Date: 2021
  • Overseas
• [Patent(Industrial Property Rights)] 2 種類の TBK1/IKKe 阻害剤を利用した核酸導入 (2020)
  • Inventor(s): 小川英知，土屋惠，渡邊賢人，平岡泰，原口徳子
  • Industrial Property Rights Holder: 小川英知，土屋惠，渡邊賢人，平岡泰，原口徳子
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2020-000088
  • Filing Date: 2020