Viral impact of hepatic non-parenchymal cells on Hepatitis B virus infected hepatocytes

• Project/Area Number: 21K07940
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 53010:Gastroenterology-related
• Research Institution: Kanazawa University
• Principal Investigator: KAWAGUCHI Kazunori 金沢大学, 医学系, 協力研究員 (90579632)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Project Status: Completed (Fiscal Year 2023)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2023: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000); Fiscal Year 2022: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2021: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: B型肝炎ウイルス / 血管内皮細胞 / 類洞内皮細胞 / 肝細胞増殖因子 / 肝非実質細胞 / 肝類洞内皮細胞 / 肝星細胞 / シングルセル解析

Outline of Research at the Start

本研究では肝組織を構成する内皮細胞、免疫担当細胞などの肝非実質細胞が、どのようなシグナル伝達を介してB型肝炎ウイルスが感染した肝細胞に対し病態変化を来すかを評価する。方法として感染した肝組織を材料としたシングルセル解析を行い、各構成細胞における遺伝子発現変化を確認する。また、肝非実質細胞と感染肝細胞の共培養を行い、肝細胞におけるウイルス動態の影響を明らかにする。さらに、肝非実質細胞がもたらす感染動態を規定する因子の同定を行う。

Outline of Final Research Achievements

Nonparenchymal cells such as endothelial cells, bile duct cells, and immune cells exist in liver tissue in addition to hepatocyte, however the interactions among these cells by hepatitis B virus infection have not been fully investigated. Previous studies we examined changes in gene expression by single cell analysis among liver component cells and found that intercellular signaling are closely related to the pathogenesis between these cells. In this study, we evaluated how vascular endothelial cells and sinusoidal endothelial cells could affect hepatitis B virus infected hepatocytes. Co-culture of endothelial cells with infected hepatocytes showed a significant increase in viral replication, suggesting the involvement of hepatocyte growth factor, and the PI3K/Akt/mTOR pathway was also found to be mediated as important signaling in hepatitis B virus infected hepatocytes.

Academic Significance and Societal Importance of the Research Achievements

核酸アナログ製剤によりB型肝炎の鎮静化がなされるが、治療後も肝細胞核内に完全二本鎖の形でウイルスが残存しており、発癌や再活性化を引き起こす。この問題を解決するために、肝組織内の構成細胞における感染による変化、細胞間の相互作用やシグナル伝達系を明確にすべきと考えた。本研究では特に、肝組織を構成する各種細胞間の作用に関わる実験的検討を行い、詳細に条件設定しながら実行し、どのような相互作用が存在するかが明らかとなった。本研究はこのウイルス残存の問題を解決し、従来の薬剤では解決できない新たな創薬にも関連するので、社会的意義のある研究とされた。

Research Products

• [Journal Article] Potential utility of l-carnitine for preventing liver tumors derived from metabolic dysfunction?associated steatohepatitis (2024)
  • Author(s): Lyu Junyan、Okada Hikari、Sunagozaka Hajime、Kawaguchi Kazunori、Shimakami Tetsuro、Nio Kouki、Murai Kazuhisa、Shirasaki Takayoshi、Yoshida Mika、Arai Kuniaki、Yamashita Tatsuya、Tanaka Takuji、Harada Kenichi、Takamura Toshinari、Kaneko Shuichi、Yamashita Taro、Honda Masao
  • Journal Title: Hepatology Communications Volume: 8 Issue: 5
  • DOI: 10.1097/hc9.0000000000000425
  • Peer Reviewed / Open Access
• [Journal Article] Serum MYCN as a predictive biomarker of prognosis and therapeutic response in the prevention of hepatocellular carcinoma recurrence (2024)
  • Author(s): Qin XY, Shirakami Y, Honda M, Yeh SH, Numata K, Lai YY, Li CL, Wei F, Xu Y, Imai K, Takai K, Chuma M, Komatsu N, Furutani Y, Gailhouste L, Aikata H, Chayama K, Enomoto M, Tateishi R, Kawaguchi K, Yamashita T, Kaneko S, Nagaoka K, Tanaka M, Sasaki Y, Tanaka Y, Baba H, Miura K, Ochi S, Masaki T, et al
  • Journal Title: International Journal of Cancer Volume: － Issue: 3 Pages: 582-594
  • DOI: 10.1002/ijc.34893
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Nucleos(t)ide analogs for hepatitis B virus infection differentially regulate the growth factor signaling in hepatocytes (2024)
  • Author(s): Shimizu Ryogo、Murai Kazuhisa、Tanaka Kensuke、Sato Yuga、Takeda Naho、Nakasyo Saki、Shirasaki Takayoshi、Kawaguchi Kazunori、Shimakami Tetsuro、Nio Kouki、Nakaya Yuki、Kagiwada Harumi、Horimoto Katsuhisa、Mizokami Masashi、Kaneko Shuichi、Murata Kazumoto、Yamashita Taro、Honda Masao
  • Journal Title: Hepatology Communications Volume: 8 Issue: 1
  • DOI: 10.1097/hc9.0000000000000351
  • Peer Reviewed / Open Access
• [Journal Article] Functional involvement of endothelial lipase in hepatitis B virus infection (2023)
  • Author(s): Shirasaki Takayoshi、Murai Kazuhisa、Ishida Atsuya、Kuroki Kazuyuki、Kawaguchi Kazunori、Wang Ying、Ito Satoru、Matsushima Kouji、Yanagawa Hiroshi、Yamashita Taro、Kaneko Shuichi、Honda Masao
  • Journal Title: Hepatology Communications Volume: 7 Issue: 9
  • DOI: 10.1097/hc9.0000000000000206
  • Peer Reviewed / Open Access
• [Journal Article] Aldo-keto reductase family 1 member B10 is regulated by nucleos(t)ide analogues for chronic hepatitis B (2023)
  • Author(s): Orita Noriaki、Kawaguchi Kazunori、Honda Masao、Shimode Tetsuhiro、Hayakawa Nozomu、Terashima Takeshi、Komura Takuya、Nishikawa Masashi、Horii Rika、Nio Kouki、Shimakami Tetsuro、Takatori Hajime、Arai Kuniaki、Sakai Yoshio、Yamashita Tatsuya、Mizukoshi Eishiro、Kaneko Shuichi、Kagaya Takashi、Yamashita Taro
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 674 Pages: 133-139
  • DOI: 10.1016/j.bbrc.2023.06.093
  • Peer Reviewed
• [Journal Article] Identification of a Transmembrane Protein Involved in Shear Stress Signaling and Hepatocarcinogenesis After a Sustained Virological Response to Hepatitis C Virus (2023)
  • Author(s): Nishikawa Masashi、Okada Hikari、Kawaguchi Kazunori、Shimakami Tetsuro、Nio Kouki、Arai Kuniaki、Yamashita Tatsuya、Sasaki Motoko、Kaneko Shuichi、Yamashita Taro、Honda Masao
  • Journal Title: Cellular and Molecular Gastroenterology and Hepatology Volume: 16 Issue: 2 Pages: 263-286
  • DOI: 10.1016/j.jcmgh.2023.04.006
  • Peer Reviewed / Open Access
• [Journal Article] Hepatitis B Virus Utilizes a Retrograde Trafficking Route via the Trans-Golgi Network to Avoid Lysosomal Degradation (2023)
  • Author(s): Li Ying-Yi、Kuroki Kazuyuki、Shimakami Tetsuro、Murai Kazuhisa、Kawaguchi Kazunori、Yamashita Taro、Kaneko Shuichi、Honda Masao
  • Journal Title: Cellular and Molecular Gastroenterology and Hepatology Volume: 15 Issue: 3 Pages: 533-558
  • DOI: 10.1016/j.jcmgh.2022.10.008
  • Peer Reviewed / Open Access
• [Journal Article] Leukocyte cell-derived chemotaxin 2 is an antiviral regulator acting through the proto-oncogene MET (2022)
  • Author(s): Shirasaki Takayoshi、Yamagoe Satoshi、Shimakami Tetsuro、Murai Kazuhisa、Imamura Ryu、Ishii Kiyo-Aki、Takayama Hiroaki、Matsumoto Yukako、Tajima-Shirasaki Natsumi、Yamashita Taro、Yoshikawa Tomoki、Takimoto Kazuhiro、Kaneko Shuichi、Matsumoto Kunio、Takamura Toshinari、Honda Masao
  • Journal Title: Nature Communications Volume: 13 Issue: 1 Pages: 3176-3176
  • DOI: 10.1038/s41467-022-30879-3
  • Peer Reviewed / Open Access
• [Journal Article] A single hepatitis B virus genome with a reporter allows the entire viral life cycle to be monitored in primary human hepatocytes (2022)
  • Author(s): Sumiyadorj Ariunaa、Murai Kazuhisa、Shimakami Tetsuro、Kuroki Kazuyuki、Nishikawa Tomoki、Kakuya Masaki、Yamada Atsumu、Wang Ying、Ishida Atsuya、Shirasaki Takayoshi、Kawase Shotaro、Li Ying‐Yi、Okada Hikari、Nio Kouki、Kawaguchi Kazunori、Honda Masao、Kaneko Shuichi
  • Journal Title: Hepatology Communications Volume: 6 Issue: 9 Pages: 2441-2454
  • DOI: 10.1002/hep4.2018
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Alterations in Hepatocellular Carcinoma-Specific Immune Responses Following Hepatitis C Virus Elimination by Direct-Acting Antivirals (2022)
  • Author(s): Li Shihui、Mizukoshi Eishiro、Kawaguchi Kazunori、Miura Miyabi、Nishino Michiko、Shimakami Tetsuro、Arai Kuniaki、Yamashita Taro、Sakai Yoshio、Yamashita Tatsuya、Honda Masao、Kaneko Shuichi
  • Journal Title: International Journal of Molecular Sciences Volume: 23 Issue: 19 Pages: 11623-11623
  • DOI: 10.3390/ijms231911623
  • Peer Reviewed / Open Access
• [Journal Article] Characterization of adipose tissue-derived stromal cells of mice with nonalcoholic fatty liver disease and their use for liver repair (2021)
  • Author(s): Yano M, Honda M, Wada T, Kaneko S, Sakai Y.（ほか17名、19番目）
  • Journal Title: Regenerative Therapy Volume: 18 Pages: 497-507
  • DOI: 10.1016/j.reth.2021.11.005
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Restorative effect of adipose tissue-derived stem cells on impaired hepatocytes through Notch signaling in non-alcoholic steatohepatitis mice (2021)
  • Author(s): Ishida Kosuke、Seki Akihiro、Kawaguchi Kazunori、Nasti Alessandro、Yamato Masatoshi、Inui Hiiro、Komura Takuya、Yamashita Taro、Arai Kuniaki、Yamashita Tatsuya、Mizukoshi Eishiro、Honda Masao、Wada Takashi、Harada Kenichi、Kaneko Shuichi、Sakai Yoshio
  • Journal Title: Stem Cell Research Volume: 54 Pages: 102425-102425
  • DOI: 10.1016/j.scr.2021.102425
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Clinical trial of autologous adipose tissue-derived regenerative (stem) cells therapy for exploration of its safety and efficacy (2021)
  • Author(s): Sakai Y, Fukunishi S, Takamura M, Kawaguchi K, Inoue O, Usui S, Takashima S, Seki A, Asai A, Tsuchimoto Y, Nasti A, Bich Ho TT, Imai Y, Yoshimura K, Murayama T, Yamashita T, Arai K, Yamashita T, Mizukoshi E, Honda M, Wada T, Harada K, Higuchi K, Kaneko S.
  • Journal Title: Regenerative Therapy Volume: 18 Pages: 97-101
  • DOI: 10.1016/j.reth.2021.04.003
  • Peer Reviewed / Open Access
• [Journal Article] DOCK11 and DENND2A play pivotal roles in the maintenance of hepatitis B virus in host cells. (2020)
  • Author(s): Hashimoto S, Shirasaki T, Yamashita T, Iwabuchi S, Suzuki Y, Takamura Y, Ukita Y, Deshimaru S, Okayama T, Ikeo K, Kuroki K, Kawaguchi K, Mizukoshi E, Matsushima K, Honda M, Kaneko S
  • Journal Title: PLoS One Volume: 16(2) Issue: 2 Pages: e0246313-e0246313
  • DOI: 10.1371/journal.pone.0246313
  • Peer Reviewed / Open Access
• [Presentation] 肝類洞内皮細胞がもたらすB型肝炎ウイルス動態の影響と亜鉛フィンガー抗ウイルス蛋白の役割 (2024)
  • Author(s): 下出哲弘，野口綾，川口和紀，丹尾幸樹，島上哲朗，村居和寿，李影奕，黒木和夫，本多政夫，山下太郎
  • Organizer: 第27回日本亜鉛栄養治療研究会学術集会
• [Presentation] 肝組織内の内皮細胞がもたらす肝細胞内のB型肝炎ウイルス複製の影響 (2023)
  • Author(s): 下出哲弘，川口和紀，山下太郎
  • Organizer: 第59回日本肝臓学会総会
• [Presentation] HGFを介した内皮細胞によるHBV感染肝細胞のウイルス複製への影響 (2023)
  • Author(s): 下出哲弘，川口和紀，本多政夫，岡田光，関晃裕，丹尾幸樹，島上哲朗，酒井佳夫，水腰英四郎，金子周一，山下太郎
  • Organizer: 第30回肝細胞研究会
• [Presentation] 脂肪組織由来間葉系幹細胞治療の非アルコール性脂肪肝炎肝細胞におけるNotchシグナルを介した修復機構 (2022)
  • Author(s): 下出哲弘、石田晃介、関晃裕、川口和紀、Alessandro Nasti、金子周一、酒井佳夫
  • Organizer: 第27回日本病態プロテアーゼ学会
• [Presentation] 肝類洞内皮細胞によるHBV感染肝細胞のAkt-mTOR経路を介したウイルス複製への影響 (2022)
  • Author(s): 下出 哲弘、川口 和紀、本多 政夫、岡田 光、村居 和寿、李 影奕、丹尾 幸樹、島上 哲朗、荒井 邦明、酒井 佳夫、山下 竜也、水腰 英四郎、金子 周一、黒木 和之、山下 太郎
  • Organizer: 第45回日本分子生物学会年会
• [Presentation] Liver sinusoidal endothelial cells affect hepatitis B virus replication in virus-infected hepatocytes via Akt-mTOR pathway (2022)
  • Author(s): Tetsuhiro Shimode, Kazunori Kawaguchi, Noriaki Orita, Kouki Nio, Hikari Okada, Kazuhisa Murai, Kouki Nio, Kuniaki Arai, Taro Yamashita, Yoshio Sakai, Tatsuya Yamashita, Eishiro Mizukoshi, Masao Honda
  • Organizer: The Liver Meeting 2022
  • Int'l Joint Research
• [Presentation] Dedicator of cytokinesis 11 is associated with the S/G2 cell cycle phase in hepatocytes with persistent hepatitis B virus infection (2022)
  • Author(s): Kazunori Kawaguchi, Taro Yamashita, Kouki Nio, Tetsuro Shimakami, Yoshio Sakai, Eishiro Mizukoshi, Masao Honda
  • Organizer: The Liver Meeting 2022
  • Int'l Joint Research
• [Presentation] 肝非実質細胞がもたらすB型肝炎ウイルス感染肝細胞におけるシグナル伝達因子の変化 (2022)
  • Author(s): 川口和紀、下出哲弘、金子周一
  • Organizer: 第108回日本消化器病学会総会
• [Presentation] Endothelial cells affect HBV replication in hepatocytes in a non-contact manner (2021)
  • Author(s): Tetsuhiro Shimode, Kazunori Kawaguchi, Masao Honda, Yoshio Sakai, Tatsuya Yamashita, Eishiro Mizukoshi, Shuichi Kaneko
  • Organizer: The Liver Meeting 2021
  • Int'l Joint Research