Novel animal models for hereditary diseases using human hepatocytes transplanted chimeric mice and vaculovirus

• Project/Area Number: 26420796
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Biofunction/Bioprocess
• Research Institution: Hiroshima University
• Principal Investigator: Chayama Hiromi 広島大学, 医歯薬保健学研究院(医), 准教授 (70572329)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: ゲノム編集 / ヒト肝細胞キメラマウス / 遺伝性肝疾患 / 遺伝性肝疾患モデル

Outline of Final Research Achievements

In this study, I established two type of CRISPR-Cas9 systems using vaculovirus vectors that work to knock out or mutate causative genes of hemochromatosis, porphyria, glycogenosis, amino acid metabolism abnormality, urea cycle disorder, constitutional jaundice, fibrotic cystic liver disease and Wilson’s disease. I transfected theses constructs into HEK293 cells and harvested vaculoviruses. However, when I inoculated HuH7 cells and HepG2 cells with the vaculoviruses, the cell growths’ were seriously inhibited. From these results, I decided to improve vaculovirus vectors. Now I confirming whether the targeted genome editing was occurred using the improved CRISPR-Cas9 systems. Further experiments are necessary. I will establish in vitro hereditary liver diseases models using human primary hepatocytes derived from human hepatocytes transplanted chimeric mice and in vivo models using human hepatocytes transplanted chimeric mice with these CRISPR-Cas9 systems.

Research Products

• [Journal Article] Development of a novel site-specific pegylated interferon beta for antiviral therapy for chronic hepatitis B. (2017)
  • Author(s): Tsuge M, Uchida T, Hiraga N, Kan H, Makokha GN, Abe-Chayama H, Miki D, Imamura M, Ochi H, Hayes CN, Shimozono R, Iwamura T, Narumi H, Suzuki T, Kainoh M, Taniguchi T, Chayama K.
  • Journal Title: Antimicrob Agents Chemother. Volume: 印刷中 Issue: 6 Pages: 61-61
  • DOI: 10.1128/aac.00183-17
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Interferon alpha treatment stimulates interferon gamma expression in type I NKT cells and enhances their antiviral effect against hepatitis C virus. (2017)
  • Author(s): Miyaki E, Hiraga N, Imamura M, Uchida T, Kan H, Tsuge M, Abe-Chayama H, Hayes CN, Makokha GN, Serikawa M, Aikata H, Ochi H, Ishida Y, Tateno C, Ohdan H, Chayama K.
  • Journal Title: PLoS One. Volume: 12 Issue: 3 Pages: e0172412-e0172412
  • DOI: 10.1371/journal.pone.0172412
  • Peer Reviewed
• [Journal Article] Usefulness of humanized cDNA-uPA/SCID mice for the study of hepatitis B virus and hepatitis C virus virology. (2017)
  • Author(s): Uchida T, Imamura M, Kan H, Hiraga N, Hayes CN, Tsuge M, Abe-Chayama H, Aikata H, Makokha GN, Miki D, Ochi H, Ishida Y, Tateno C, Chayama K.
  • Journal Title: J Gen Virol. Volume: 印刷中 Issue: 5 Pages: 1040-1047
  • DOI: 10.1099/jgv.0.000726
  • Peer Reviewed
• [Journal Article] Antiviral effects of anti-HBs immunoglobulin and vaccine on HBs antigen seroclearance for chronic hepatitis B infection. (2016)
  • Author(s): Tsuge M, Hiraga N, Uchida T, Kan H, Miyaki E, Masaki K, Ono A, Nakahara T, Abe-Chayama H, Zhang Y, Naswa MG, Kawaoka T, Miki D, Imamura M, Kawakami Y, Aikata H, Ochi H, Hayes CN, Chayama K.
  • Journal Title: J Gastroenterol. Volume: 51 Pages: 1073-1080
  • Peer Reviewed
• [Journal Article] Highly multiplexed CRISPR-Cas9-nuclease and Cas9-nickase vectors for inactivation of hepatitis B virus. (2016)
  • Author(s): Sakuma T, Masaki K, Abe-Chayama H, Mochida K, Yamamoto T, Chayama K.
  • Journal Title: Genes Cells. Volume: 21 Issue: 11 Pages: 1253-1262
  • DOI: 10.1111/gtc.12437
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Elimination of HCV via a non-ISG-mediated mechanism by vaniprevir andBMS-788329 combination therapy in human hepatocyte chimeric mice. (2016)
  • Author(s): Uchida T, Hiraga N, Imamura M, Yoshimi S, Kan H, Miyaki E, Tsuge M, Abe H, Hayes CN, Aikata H, Ishida Y, Tateno C, Ellis JD, Chayama K.
  • Journal Title: Virus Res. Volume: 213 Issue: 1 Pages: 62-68
  • DOI: 10.1016/j.jinf.2015.09.038
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Human Cytotoxic T Lymphocyte-Mediated Acute Liver Failure and Rescue by Immunoglobulin in Human Hepatocyte Transplant TK-NOG Mice. (2015)
  • Author(s): Uchida T., Hiraga N., Imamura M., Tsuge M., Abe H., Hayes C.N., Aikata H., Ishida Y., Tateno C., Yoshizato K., Ohdan H., Murakami K. and Chayama K.
  • Journal Title: Journal of Virology Volume: 89 Issue: 19 Pages: 10087-96
  • DOI: 10.1128/jvi.01126-15
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Rapid, Sensitive, and Accurate Evaluation of Drug Resistant Mutant (NS5A-Y93H) Strain Frequency in Genotype 1b HCV by Invader Assay (2015)
  • Author(s): Satoshi Yoshimi, Hidenori Ochi, Eisuke Murakami, Takuro Uchida, Hiromi Kan, Sakura Akamatsu, C. Nelson Hayes, Hiromi Abe, Daiki Miki, Nobuhiko Hiraga, Michio Imamura, Hiroshi Aikata, Kazuaki Chayama.
  • Journal Title: PLOS ONE Volume: 10 Issue: 6 Pages: e0130022-e0130022
  • DOI: 10.1371/journal.pone.0130022
  • Peer Reviewed / Open Access
• [Journal Article] Long term persistence of NS5A inhibitor-resistant hepatitis C virus in patients who failed daclatasvir and asunaprevir therapy. (2015)
  • Author(s): Yoshimi S, Imamura M, Murakami E, Hiraga N, Tsuge M, Kawakami Y, Aikata H, Abe H, Hayes CN, Sasaki T, Ochi H, Chayama K.
  • Journal Title: J Med Virol. Volume: 87 Issue: 11 Pages: 1913-1920
  • DOI: 10.1002/jmv.24255
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Novel Robust in Vitro Hepatitis B Virus Infection Model Using Fresh Human Hepatocytes Isolated from Humanized Mice. (2015)
  • Author(s): Ishida Y, Yamasaki C, Yanagi A, Yoshizane Y, Fujikawa K, Watashi K, Abe H, Wakita T, Hayes CN, Chayama K, Tateno C.
  • Journal Title: Am J Pathol. Volume: 185 Issue: 5 Pages: 1275-1285
  • DOI: 10.1016/j.ajpath.2015.01.028
  • Peer Reviewed
• [Journal Article] Association between variants in the interferon lambda 4 locus and substitutions in the hepatitis C virus non-structural protein 5A. (2015)
  • Author(s): Akamatsu S, Hayes CN, Ochi H, Uchida T, Kan H, Murakami E, Abe H, Tsuge M, Miki D, Akiyama R, Hiraga N, Imamura M, Aikata H, Kawaoka T, Kawakami Y, Chayama K.
  • Journal Title: J Hepatol. Volume: 63 Issue: 3 Pages: 554-563
  • DOI: 10.1016/j.jhep.2015.03.033
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Differences in serum microRNA profiles in hepatitis B and C virus infection. (2015)
  • Author(s): Akamatsu S, Hayes CN, Tsuge M, Miki D, Akiyama R, Abe H, Ochi H, Hiraga N, Imamura M, Takahashi S, Aikata H, Kawaoka T, Kawakami Y, Ohishi W, Chayama K.
  • Journal Title: J Infect. Volume: 70 Issue: 3 Pages: 273-287
  • DOI: 10.1016/j.jinf.2014.10.017
  • Peer Reviewed
• [Journal Article] The effects of bisphosphonate zoledronic acid in hepatocellular carcinoma, depending on mevalonate pathway. (2015)
  • Author(s): Honda Y, Takahashi S, Zhang Y, Ono A, Murakami E, Shi N, Kawaoka T, Miki D, Tsuge M, Hiraga N, Abe H, Ochi H, Imamura M, Aikata H, Chayama K.
  • Journal Title: J Gastroenterol Hepatol. Volume: 30 Issue: 3 Pages: 619-627
  • DOI: 10.1111/jgh.12715
  • Peer Reviewed / Open Access
• [Journal Article] New insight into the enhanced effect of pegylated interferon-α. (2014)
  • Author(s): Abe H, Hayes CN, Chayama K.
  • Journal Title: Hepatology. Volume: 60 Issue: 4 Pages: 1435-1437
  • DOI: 10.1002/hep.27269
  • Peer Reviewed / Open Access
• [Journal Article] Emergence of resistant variants detected by ultra-deep sequencing after asunaprevir and daclatasvir combination therapy in patients infected with hepatitis C virus genotype 1. (2014)
  • Author(s): Kosaka K, Imamura M, Hayes CN, Abe H, Hiraga N, Yoshimi S, Murakami E, Kawaoka T, Tsuge M, Aikata H, Miki D, Ochi H, Matsui H, Kanai A, Inaba T, Chayama K.
  • Journal Title: J Viral Hepat. Volume: 22 Issue: 2 Pages: 158-165
  • DOI: 10.1111/jvh.12271
  • Peer Reviewed / Open Access
• [Journal Article] Ribavirin dose reduction during telaprevir/ribavirin/peg-interferon therapy overcomes the effect of the ITPA gene polymorphism. (2014)
  • Author(s): Akamatsu S, Hayes CN, Tsuge M, Murakami E, Hiraga N, Abe H, Miki D, Imamura M, Ochi H, Chayama K; Hiroshima Liver Study Group.
  • Journal Title: J Viral Hepat. Volume: 22 Issue: 2 Pages: 166-174
  • DOI: 10.1111/jvh.12275
  • Peer Reviewed
• [Journal Article] Human microRNA hsa-miR-1231 suppresses hepatitis B virus replication by targeting core mRNA. (2014)
  • Author(s): Kohno T, Tsuge M, Murakami E, Hiraga N, Abe H, Miki D, Imamura M, Ochi H, Hayes CN, Chayama K.
  • Journal Title: J Viral Hepat. Volume: 21 Issue: 4 Pages: 369-375
  • DOI: 10.3851/imp2777
  • Peer Reviewed
• [Presentation] Analysis of the effect on HBV by genome editing using CRISPR/Cas9 system (2016)
  • Author(s): Keiichi Masaki, Hiromi Abe, Tetsushi Sakuma, Masataka Tsuge, Michio Imamura, Nelson Hayes, Hiroshi Aikata, Takashi Yamamoto, Kazuaki Chayama
  • Organizer: AASLD2016
  • Place of Presentation: アメリカ・ボストン
  • Year and Date: 2016-11-11
  • Int'l Joint Research
• [Presentation] Analysis of the effect on HBV life cycle by HBV genome editing using two different CRISPR/Cas9 systems. (2015)
  • Author(s): Abe H, Masaki K, Sakuma T, Tsuge M, Hiraga N, Imamura M, Hayes CN, Aikata H, Yamamoto T, Chayama K.
  • Organizer: The 66th AASLD Annual Meeting
  • Place of Presentation: San Francisco, USA.
  • Year and Date: 2015-11-13
  • Int'l Joint Research
• [Presentation] HCV感染におけるIFNL4の多型の違いによるIFN-λsの発現量とインターフェロン投与前後のインターフェロン誘導遺伝子の発現量の関係 (2014)
  • Author(s): 阿部弘美, 三木大樹, 平賀伸彦, 今村道雄, 柘植雅貴, 小林知樹, 河岡友和, 川上由育, 相方浩, 越智秀典, 茶山一彰
  • Organizer: 第50回日本肝臓学会総会
  • Place of Presentation: 東京
  • Year and Date: 2014-05-29 – 2014-05-30
• [Remarks] 広島大学大学院医歯薬保健学研究科消化器・代謝内科
  • URL: http://shounai.hiroshima-u.ac.jp/
• [Remarks] 広島大学大学院医歯薬保健学研究院応用生命科学部門消化器・代謝内科学ホームページ
  • URL: http://home.hiroshima-u.ac.jp/naika1/