Identification of long non-coding RNAs that regulate HBV replication

• Project/Area Number: 18K07161
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 49060:Virology-related
• Research Institution: Fujita Health University (2020); National Center for Global Health and Medicine (2018-2019)
• Principal Investigator: Nishitsuji Hironori 藤田医科大学, 医学部, 講師 (20573661)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2019: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2018: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: HBV / lncRNA / HBV Core / HBV X / long non-coding RNA

Outline of Final Research Achievements

In this study, I investigated the roles of long non-coding RNA in HBV replication. HepG2 cells were expressed with HA- and myc-tagged HBV X protein. Cell lysates were immunoprecipitated with anti-HA and anti-myc antibodies. Immunocomplex were subjected to next generation sequence to identify lncRNAs bound to HBV X protein. I found that miRNA acts as suppressor of HBV replication through degradation of HBV X protein.

Academic Significance and Societal Importance of the Research Achievements

今回、Xタンパク質と結合するmiRNAを同定した。この同定したmiRNAはHBV Xタンパク質と結合し、Xタンパク質の機能を阻害した。さらにこのmiRNAはHBV感染も阻害した。このmiRNAはXタンパク質と結合することにより、Xタンパク質を不安定化した。Xタンパク質は、HBVの転写を制御していることから、このmiRNAがHBVの潜伏化に関与している可能が示唆される。また、このmiRNAを人工的に制御できれば、HBV阻害薬のターゲットととして期待される。

Research Products

• [Journal Article] Identification of Two Critical Neutralizing Epitopes in the Receptor Binding Domain of Hepatitis B Virus preS1 (2020)
  • Author(s): Yato Keigo、Onodera Taishi、Matsuda Mami、Moriyama Saya、Fujimoto Akira、Watashi Koichi、Aizaki Hideki、Tanaka Tomohisa、Moriishi Kohji、Nishitsuji Hironori、Shimotohno Kunitada、Tamura Koji、Takahashi Yoshimasa、Wakita Takaji、Muramatsu Masamichi、Kato Takanobu、Suzuki Ryosuke
  • Journal Title: Journal of Virology Volume: 95 Issue: 5
  • DOI: 10.1128/jvi.01680-20
  • Peer Reviewed / Open Access
• [Journal Article] Modulation of hepatitis B virus infection by epidermal growth factor secreted from liver sinusoidal endothelial cells (2020)
  • Author(s): Shin-Wei Chen, Misao Himeno, Yuta Koui, Masaya Sugiyama, Hironori Nishitsuji, Masashi Mizokami, Kunitada Shimotohno, Atsushi Miyajima, Taketomo Kido
  • Journal Title: Scientific Reports Volume: 10(1):14349 Issue: 1 Pages: 1-10
  • DOI: 10.1038/s41598-020-71453-5
  • Peer Reviewed / Open Access
• [Journal Article] Orchestration of Intracellular Circuits by G Protein-Coupled Receptor 39 for Hepatitis B Virus Proliferation. (2020)
  • Author(s): Goto K, Nishitsuji H, Sugiyama M, Nishida N, Mizokami M, Shimotohno K.
  • Journal Title: International Journal of Molecular Sciences Volume: 21 Issue: 16 Pages: 5661-5661
  • DOI: 10.3390/ijms21165661
  • Peer Reviewed / Open Access
• [Journal Article] N‐terminal PreS1 Sequence Regulates Efficient Infection of Cell Culture‐generated Hepatitis B Virus (2020)
  • Author(s): Murayama A、Yamada N、Osaki 、Shiina 、Aly H H、Iwamoto M、Tsukuda S、Watashi K、Matsuda M、Suzuki R、Tanaka T、Moriishi K、Suzuki T、Nishitsuji H、Sugiyama M、Mizokami M、Shimotohno K、Wakita T、Muramatsu M、Liang T J、Kato T
  • Journal Title: Hepatology Volume: 「－」 Issue: 2 Pages: 520-532
  • DOI: 10.1002/hep.31308
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Engineering Cellular Biosensors with Customizable Antiviral Responses Targeting Hepatitis B Virus (2020)
  • Author(s): Matsunaga Satoko、Jeremiah Sundararaj S.、Miyakawa Kei、Kurotaki Daisuke、Shizukuishi Sayaka、Watashi Koichi、Nishitsuji Hironori、Kimura Hirokazu、Tamura Tomohiko、Yamamoto Naoki、Shimotohno Kunitada、Wakita Takaji、Ryo Akihide
  • Journal Title: iScience Volume: 23 Issue: 3 Pages: 100867-100867
  • DOI: 10.1016/j.isci.2020.100867
  • Peer Reviewed / Open Access
• [Journal Article] Anti‐viral effects of interferon‐λ3 on hepatitis B virus infection in cell culture (2020)
  • Author(s): Yamada Norie、Murayama Asako、Shiina Masaaki、Aly Hussein Hassan、Iwamoto Masashi、Tsukuda Senko、Watashi Koichi、Tanaka Tomohisa、Moriishi Kohji、Nishitsuji Hironori、Sugiyama Masaya、Mizokami Masashi、Shimotohno Kunitada、Muramatsu Masamichi、Murata Kazumoto、Kato Takanobu
  • Journal Title: Hepatology Research Volume: 50 Issue: 3 Pages: 283-291
  • DOI: 10.1111/hepr.13449
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] A new hepatoma cell line exhibiting high susceptibility to hepatitis B virus infection (2019)
  • Author(s): Ueda Youki、Gu Weilin、Dansako Hiromichi、Nishitsuji Hironori、Satoh Shinya、Shimotohno Kunitada、Kato Nobuyuki
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 515 Issue: 1 Pages: 156-162
  • DOI: 10.1016/j.bbrc.2019.05.126
  • Peer Reviewed
• [Journal Article] Serum miRNAs Predicting Sustained HBs Antigen Reduction 48 Weeks after Pegylated Interferon Therapy in HBe Antigen-Negative Patients (2018)
  • Author(s): Fujita Koji、Mimura Shima、Iwama Hisakazu、Nakahara Mai、Oura Kyoko、Tadokoro Tomoko、Nomura Takako、Tani Joji、Yoneyama Hirohito、Morishita Asahiro、Oryu Makoto、Himoto Takashi、Nishitsuji Hironori、Shimotohno Kunitada、Omata Masao、Masaki Tsutomu
  • Journal Title: International Journal of Molecular Sciences Volume: 19 Issue: 7 Pages: 1940-1940
  • DOI: 10.3390/ijms19071940
  • Peer Reviewed
• [Journal Article] Development of a cell-based assay to identify hepatitis B virus entry inhibitors targeting the sodium taurocholate cotransporting polypeptide (2018)
  • Author(s): Miyakawa Kei、Matsunaga Satoko、Yamaoka Yutaro、Dairaku Mina、Fukano Kento、Kimura Hirokazu、Chimuro Tomoyuki、Nishitsuji Hironori、Watashi Koichi、Shimotohno Kunitada、Wakita Takaji、Ryo Akihide
  • Journal Title: Oncotarget Volume: 9 Issue: 34 Pages: 23681-23694
  • DOI: 10.18632/oncotarget.25348
  • Peer Reviewed / Open Access