Integrating multiple omics analysis for understanding the pathological mechanism of chronic hepatitis B virus infection and identifying potential molecular targets using humanized chimeric mice

2017 Fiscal Year Annual Research Report

• Project/Area Number: 15K06810
• Research Institution: Jikei University School of Medicine
• Principal Investigator: 坪田 昭人 東京慈恵会医科大学, 医学部, 教授 (90322643)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: B型肝炎 / B型肝炎ウイルス / トランスクリプトーム解析 / ヒト化肝臓キメラマウス

Outline of Annual Research Achievements

核酸アナログ体は、逆転写酵素が関与するB型肝炎ウイルス(HBV)の複製過程を効果的に抑制するが、その他の増殖過程には作用せず、感染肝細胞からHBVを完全には排除できない。新たな作用機序をもつ薬剤が必要である。本研究では、低コストで安定した動物モデルを確立し、そのモデルを用いてHBVが急性感染から慢性化するまでの肝組織内トランスクリプトームを網羅的に解析することにより慢性化の機序を解明、新規治療への基礎的検討を行った。
HBVに最適な培養系はなく、臨床検体の入手には限界がある。臨床とin vitroの橋渡しとしてヒト肝細胞で置換されたキメラマウスが現時点で適切である。我々は、TK-NOGマウスに独自のプロトコールと移植法を施し、低コストで置換率の高いヒト化肝臓キメラマウスを作製した。この動物モデルを用いて、HBV感染の超急性感染から慢性化に移行するまでの網羅的トランスクリプトーム解析を行った。その結果、高発現遺伝子群より圧倒的に低発現遺伝子群が多く、前者にはTLR/アポトーシス関連遺伝子が含まれていた。一方、validationにより経過を通じて高発現していた3つのmicroRNAを同定した。Primaryなヒト肝細胞系で、これらのmicroRNAを阻害するとHBVの感染効率が低下した。HBV感染肝細胞株で候補microRNAのcounterpartであるmRNAを確定し、そのmRNAがHBV増殖に影響することも明らかになってきた。プロテオミクス解析やメタボロミクス解析では、リポ蛋白系の変動が主であった。
HBV侵入直後の感染超初期で肝細胞にどのような変化が起きているか、十分にわかっていない。感染成立機序を解明することで感染阻止の分子標的を特定でき得ると考えられ、新たな治療の発展につながる可能性がある。HBVは自身に有利な環境を作りながら宿主に適応するものと考えられた。

Research Products

• [Journal Article] Efficacy and safety of ledipasvir/sofosbuvir for genotype 1b chronic hepatitis C patients with moderate renal impairment (2018)
  • Author(s): Okubo T, Atsukawa M, Tsubota A, Toyoda H, Shimada N, Abe H, Kato K, Hayama K, Arai T, Nakagawa-Iwashita A, Itokawa N, Kondo C, Kawamoto C, Iio E, Tanaka Y, Kumada T, Iwakiri K.
  • Journal Title: Hepatol Int Volume: 12 Pages: 133-142
  • DOI: 10.1007/s12072-018-9859-9
  • Peer Reviewed
• [Journal Article] Hepatocellular adenoma in a woman who was undergoing testosterone treatment for gender identity disorder (2018)
  • Author(s): Kato K, Abe H, Hanawa N, Fukuzawa J, Matsuo R, Yonezawa T, Itoh S, Sato Y, Ika M, Shimizu S, Endo S, Hano H, Izu A, Sugitani M, Tsubota A
  • Journal Title: Clin J Gastroenterol Volume: Mar 27 Pages: ―
  • DOI: 10.1007/s12328-018-0854-4
  • Peer Reviewed / Open Access
• [Journal Article] Development of a New Conditionally Immortalized Human Liver Sinusoidal Endothelial Cells (2018)
  • Author(s): Zhu M, Koibuchi A, Ide H, Morio H, Shibuya M, Kamiichi A, Tsubota A, Anzai N, Akita H, Chiba K, Furihata T.
  • Journal Title: Biol Pharm Bull Volume: 41 Pages: 440-444
  • DOI: 10.1248/bpb.b17-00661
  • Peer Reviewed
• [Journal Article] Efficacy and safety of ombitasvir/paritaprevir/ritonavir combination therapy for genotype 1b chronic hepatitis C patients complicated with chronic kidney disease (2018)
  • Author(s): Arai T, Atsukawa M, Tsubota A, Ikegami T, Shimada N, Kato K, Abe H, Okubo T, Itokawa N, Kondo C, Mikami S, Asano T, Chuganji Y, Matsuzaki Y, Toyoda H, Kumada T, Iio E, Tanaka Y, Iwakiri K.
  • Journal Title: Hepatol Res Volume: 印刷中 Pages: 印刷中
  • DOI: 10.1111/hepr.13058
  • Peer Reviewed
• [Journal Article] Serum Wisteria floribunda agglutinin-positive Mac-2 binding protein more reliably distinguishes liver fibrosis stages in non-alcoholic fatty liver disease than serum Mac-2 binding protein (2017)
  • Author(s): Atsukawa M, Tsubota A, Okubo T, Arai T, Nakagawa A, Itokawa N, Kondo C, Kato K, Hatori T, Hano H, Oikawa T, Emoto N, Abe M, Kage M, Iwakiri K.
  • Journal Title: Hepatol Res Volume: 印刷中 Pages: 印刷中
  • DOI: 10.1111/hepr.13046
  • Peer Reviewed
• [Journal Article] Analysis of factors predicting the response to tolvaptan in patients with liver cirrhosis and hepatic edema (2017)
  • Author(s): Atsukawa M, Tsubota A, Kato K, Abe H, Shimada N, Asano T, Ikegami T, Koeda M, Okubo T, Arai T, Nakagawa-Iwashita A, Yoshida Y, Hayama K, Itokawa N, Kondo C, Chuganji Y, Matsuzaki Y, Iwakiri K.
  • Journal Title: J Gastroenterol Hepatol Volume: 印刷中 Pages: 印刷中
  • DOI: 10.1111/jgh.14047
  • Peer Reviewed
• [Journal Article] Disorders in Hepatic Copper Secretion: Wilson's Disease and Pleomorphic Syndromes (2017)
  • Author(s): Lalioti V, Tsubota A, Sandoval IV.
  • Journal Title: Semin Liver Dis Volume: 37 Pages: 175-188
  • DOI: 10.1055/s-0037-1602764
  • Peer Reviewed
• [Journal Article] Effectiveness and safety of community-based treatment with sofosbuvir plus ribavirin for elderly patients with genotype 2 chronic hepatitis C (2017)
  • Author(s): Atsukawa M, Tsubota A, Kondo C, Shimada N, Abe H, Kato K, Okubo T, Arai T, Itokawa N, Iio E, Tanaka Y, Iwakiri K.
  • Journal Title: Dig Liver Dis Volume: 49 Pages: 1029-1035
  • DOI: 10.1016/j.dld.2017.04.012
  • Peer Reviewed
• [Journal Article] Efficacy and safety of ombitasvir/paritaprevir/ritonavir in dialysis patients with genotype 1b chronic hepatitis C (2017)
  • Author(s): Atsukawa M, Tsubota A, Koushima Y, Ikegami T, Watanabe K, Shimada N, Sato S, Kato K, Abe H, Okubo T, Arai T, Itokawa N, Kondo C, Mikami S, Asano T, Chuganji Y, Matsuzaki Y, Iwakiri K.
  • Journal Title: Hepatol Res Volume: 47 Pages: 1429-1437
  • DOI: 10.1111/hepr.12910
  • Peer Reviewed
• [Journal Article] Clinical evaluation of sofosbuvir/ledipasvir in patients with chronic hepatitis C genotype 1 with and without prior daclatasvir/asunaprevir therapy (2017)
  • Author(s): Iio E, Shimada N, Takaguchi K, Senoh T, Eguchi Y, Atsukawa M, Tsubota A, Abe H, Kato K, Kusakabe A, Miyaki T, Matsuura K, Matsunami K, Shinkai N, Fujiwara K, Nojiri S, Tanaka Y.
  • Journal Title: Hepatol Res Volume: 47 Pages: 1308-1316
  • DOI: 10.1111/hepr.12898
  • Peer Reviewed
• [Journal Article] Retreatment with sofosbuvir, ledipasvir, and add-on ribavirin for patients who failed daclatasvir and asunaprevir combination therapy (2017)
  • Author(s): Suda G, Ogawa K, Yamamoto Y, Katagiri M, Furuya K, Kumagai K, Konno J, Kimura M, Kawagishi N, Ohara M, Umemura M, Ito J, Izumi T, Nakai M, Sho T, Natsuizaka M, Morikawa K, Tsubota A, Shimada N, Iio E, Tanaka Y, Sakamoto N; NORTE Study Group.
  • Journal Title: J Gastroenterol Volume: 52 Pages: 1122-1129
  • DOI: 10.1007/s00535-017-1328-z
  • Peer Reviewed
• [Journal Article] Daclatasvir and asunaprevir for genotype 1b chronic hepatitis C patients with chronic kidney disease (2017)
  • Author(s): Kondo C, Atsukawa M, Tsubota A, Shimada N, Abe H, Asano T, Yoshizawa K, Okubo T, Chuganji Y, Aizawa Y, Iio E, Tanaka Y, Iwakiri K.
  • Journal Title: Hepatol Res Volume: 47 Pages: 1165-1173
  • DOI: 10.1111/hepr.12879
  • Peer Reviewed
• [Journal Article] Clinical Significance of Two Real-Time PCR Assays for Chronic Hepatitis C Patients Receiving Protease Inhibitor-Based Therapy (2017)
  • Author(s): Inoue T, Hmwe SS, Shimada N, Kato K, Ide T, Torimura T, Kumada T, Toyoda H, Tsubota A, Takaguchi K, Wakita T, Tanaka Y.
  • Journal Title: PLoS One Volume: 12 Pages: e0170667
  • DOI: 10.1371/journal.pone.0170667
  • Peer Reviewed / Open Access
• [Journal Article] C-Arm Cone Beam Computed Tomography Guidance for Radiofrequency Ablation in Hepatocellular Carcinoma (2017)
  • Author(s): Kato K, Abe H, Ika M, Yonezawa T, Sato Y, Hanawa N, Shimizu S, Endo S, Matsuo R, Tsubota A.
  • Journal Title: Oncology Volume: 92 Pages: 142-152
  • DOI: 10.1159/000453665
  • Peer Reviewed
• [Journal Article] Human organic anion transporter 2 is an entecavir, but not tenofovir, transporter (2017)
  • Author(s): Furihata T, Morio H, Zhu M, Suzuki Y, Ide H, Tsubota A, Fu Z, Anzai N, Chiba K.
  • Journal Title: Drug Metab Pharmacokinet Volume: 32 Pages: 116-119
  • DOI: 10.1016/j.dmpk.2016.09.004
  • Peer Reviewed
• [Presentation] Ultrastructure of hepatocytes in chronic hepatitis C patients who achieve a sustained virological response (2017)
  • Author(s): Haruyo Aoyagi, Hiroko Iijima, Francesc Puig-Basagoiti, Zheng Xin, Yu Ting Kao, Gewaid E. Hossam, Takuma Zaitsu, Mami Matsuda, Koichi Watashi, Ryosuke Suzuki, Takahiro Masaki, Noritomo Shimada, Keizo Kato, Akihito Tsubota, Ayako Mimata, Yuriko Sakamaki, Shizuko Ichinose, Kenjiro Wake, Takaji Wakita, Hideki Aizaki
  • Organizer: APASL 2017 (26th)
  • Int'l Joint Research
• [Presentation] Abnormal Hepatocellular Organelles Remain to be Observed in Sustained Virological Response Patients (2017)
  • Author(s): Haruyo Aoyagi, Hiroko Iijima, Francesc Puig-Basagoiti, Zheng Xin, Yu Ting Kao, Gewaid E. Hossam, Takuma Zaitsu, Mami Matsuda, Koichi Watashi, Ryosuke Suzuki, Takahiro Masaki, Akihito Tsubota, Mimata Ayako, Yuriko Sakamaki, Shizuko Ichinose, Kenjiro Wake, Takaji Wakita, Hideki Aizaki
  • Organizer: APASL Single Topic Conference
  • Int'l Joint Research
• [Presentation] SVR後の肝臓のオルガネラ異常の解析 (2017)
  • Author(s): 青柳東代, 飯島尋子, Francesc Puig-Basagoiti, Zheng Xin, Yu Ting Kao, Gewaid E. Hossam, 在津拓馬, 松田麻未, 渡士幸一, 鈴木亮介, 政木隆博, 島田紀朋, 加藤慶三, 坪田昭人, 三又絢子, 酒巻有里子, 市野瀬志津子, 和氣健二郎, 脇田隆字, 相崎英樹.
  • Organizer: 第27回抗ウイルス療法学会総会
• [Presentation] Abnormal hepatocellular organelles remain to be observed in sustained virological response patients (2017)
  • Author(s): Aoyagi H, Iijima H, Puig-Basagoiti F, Xin Z, Kao Y-T, Hossam GE, Zaitsu T, Matsuda M, Watashi K, Suzuki R, Masaki T, Aizawa N, Shimada N, Kato K, Tsubota A, Mimata A, Sakamaki Y, Ichinose S, Wake K, Wakita T, Aizaki H
  • Organizer: 第65回日本ウイルス学会学術集会
• [Presentation] A novel device for estimating olfactory threshold for differential diagnosis of parkinsonian syndrome: a pilot study (2017)
  • Author(s): Omoto S, Mori E, Fujioka K, Tsubota A, Kita J, Kinoshita M, Matsuno H, Sakai K, Toyoda C, Iguchi Y
  • Organizer: XXIII World Congress of Neurology
  • Int'l Joint Research