The aa15-17 amino acid sequence in the terminal protein domain of HBV polymerase as a viral factor affect-ing in vivo as well as in vitro replication activity of the virus.

• Project/Area Number: 25461010
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Gastroenterology
• Research Institution: Saitama Medical University
• Principal Investigator: Mochida Satoshi 埼玉医科大学, 医学部, 教授 (20219968)
• Co-Investigator(Kenkyū-buntansha): NAKAYAMA Nobuaki 埼玉医科大学, 医学部, 准教授 (40292015) ; Sugawara Michiko 埼玉医科大学, 医学部, 講師 (20406458)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: HBV / HCV / polymerase / terminal protein domain / nucleoside / nucleotide analogs / cyclingprobereal-timePCR / 核酸アナログ / B型慢性肝炎 / ポリメラーゼ / Huh7

Outline of Final Research Achievements

In patients with HBV infection, serum HBV-DNA levels increased more rapidly after discontinuation of nu-cleos(t)ide analogs in those with HBV showing DDE motif at aa15-17 in terminal protein domain of poly-merase than in those with HBV in which either of amino acids of the motif was replaced to neutral amino ac-ids. Thus, HBV strains manifesting different sequences at aa15-17 were transfected into Huh7 cells, and HBV-DNA levels in the culture medium were measured. Consequently, the motif was shown to be responsible for replication activity of HBV in vitro as well as in vivo. To develop a simple assay system to determine amino acids sequence of the motif, which may be useful to predict clinical features of patients with HBV, we adopted cycling probe real-time PCR. We established the system to determine NS5A-Y93H mutation in HCV, and the project to apply this method for the motif in HBV is no on progress.

Research Products

• [Journal Article] Development of rare RAVs that are extremely tolerant against NS5A inhibitors during daclatasvir/asunaprevir therapy via a two-hit mechanism. (2016)
  • Author(s): Uchida Y, Kouyama J, Naiki K, Sugawara K, Inao M, Imai Y, Nakayama N, Mochida S.
  • Journal Title: Hepatology Research Volume: Feb 15 Issue: 12 Pages: 1234-1246
  • DOI: 10.1111/hepr.12673
  • Peer Reviewed
• [Journal Article] Nationwide prospective and retrospective surveys for hepatitis B virus reactivation during immunosuppressive therapies. (2016)
  • Author(s): Mochida S, Nakao M, Nakayama N, Uchida Y, Nagoshi S, Ido A, Mimura T, Harigai M, Kaneko H, Kobayashi H, Tsuchida T, Suzuki H, Ura N, Nakamura Y, Bessho M, Dan K, Kusumoto S, Sasaki Y, Fujii H, Suzuki F, Ikeda K, Yamamoto K, Takikawa H, Tsubouchi H, Mizokami M.
  • Journal Title: Journal of Gastroenterology Volume: Feb 1
  • Peer Reviewed
• [Journal Article] Significance of variants associated with resistance to NS5A inhibitors in Japanese patients with genotype 1b hepatitis C virus infection as evaluated using cycling-probe real-time PCR combined with direct sequencing. (2016)
  • Author(s): Uchida Y, Kouyama J, Naiki K, Sugawara K, Ando S, Nakaoa M, Motoya D, Inao M, Imai Y, Nakayama N, Mochida S.
  • Journal Title: Journal of Gastroenterology Volume: 51 Issue: 3 Pages: 260-270
  • DOI: 10.1007/s00535-015-1106-8
  • Peer Reviewed
• [Journal Article] Revised criteria for classification of the etiologies of acute liver failure and late-onset hepatic failure in Japan: A report by the Intractable Hepato-Biliary Diseases Study Group of Japan in 2015. (2015)
  • Author(s): Mochida S, Nakayama N, Ido A, Takikawa Y, Yokosuka O, Sakaida I, Moriwaki H, Genda T, Takikawa H.
  • Journal Title: Hepatology Research Volume: Nov 28 Issue: 5 Pages: 369-371
  • DOI: 10.1111/hepr.12626
  • Peer Reviewed
• [Journal Article] Drug-induced immunoallergic hepatitis during combination therapy with daclatasvir and asunaprevir. (2015)
  • Author(s): Ｆujii Y, Uchida Y, Mochida S
  • Journal Title: Hepatology Volume: 61 Issue: 1 Pages: 400-401
  • DOI: 10.1002/hep.27559
  • Peer Reviewed
• [Journal Article] Multicenter prospective study to optimize the efficacy of triple therapy with telaprevir in patients with genotype 1b HCV infection according to an algorithm based on the drug adherence, IL28B gene allele and viral response: the AG & RGT Trial. (2015)
  • Author(s): Sugawara K, Koushima Y, Inao M, Nakayama N, Nagoshi S, Yakabi K, Tamano M, Asabe S, Mishikawa K, Harada Y, Sekine C, Fukuya Y, Funtny J, Hashimoto Y, Mochida S
  • Journal Title: Hepatol Res Volume: in press Issue: 11 Pages: 1091-1099
  • DOI: 10.1111/hepr.12475
  • Peer Reviewed
• [Journal Article] Prospective randomized study comparing enhancement of the antiviral efficacy between vitamin D3 (cholecalciferol) and 1α(OH) D3 (alfacalcidol) in combination with pegylated interferon plus ribavirin for patients with chronic hepatitis C. (2015)
  • Author(s): Mizuno Y, Nakayama N, Inao M, Funyu J, Asabe S, Tomita K, Nishikawa K, Hosoda Y, Tanaka M, Hashimoto Y, Takabayashi H, Yakabi K, Koshima K, Mochida S
  • Journal Title: J Gastroenterol Hepatol Volume: in press
  • Peer Reviewed
• [Journal Article] A novel simple assay system to quantify the percent HCV-RNA Levels of NS5A Y93H mutant strains and Y93 wild-type strains relative to the total HCV-RNA levels to determine the indication for antiviral therapy with NS5A inhibitors. (2014)
  • Author(s): 12.Uchida Y, Kouyama J-I, Naiki K, Mochida S
  • Journal Title: PloS One Volume: 9
  • Peer Reviewed
• [Journal Article] Genome-wide association study identifies a PSMD3 variant associated with neutropenia in inter-feron-based therapy for chronic hepatitis C. (2014)
  • Author(s): Iio E, Mochida S,et al. (計31名中18番目)
  • Journal Title: Hum Genet Volume: 17
  • Peer Reviewed
• [Journal Article] Classification of the etiologies of acute liver failure in Japan: A report by the Intractable Hepato-Biliary Diseases Study Group of Japan. (2014)
  • Author(s): Mochida S, Takikawa Y, Nakayama N, Oketani M, Naiki T, Yamagishi Y, Fujiwara K, Ichida T, Tsubouchi H.
  • Journal Title: Hepatol Res Volume: in press Issue: 4 Pages: 365-367
  • DOI: 10.1111/hepr.12295
  • NAID: 130003393966
  • Peer Reviewed
• [Journal Article] New susceptibility and resistance HLA-DP alleles to HBV-related diseases identified by a trans-ethnic association study in Asia. (2014)
  • Author(s): Nishida N, Sawai H, Kashiwase K, Minami M, Sugiyama M, Seto WK, Yuen MF, Posuwan N, Poovorawan Y, Ahn SH, Han KH, Matsuura K, Tanaka Y, Kurosaki M, Asahina Y, Izumi N, Kang JK, （省略）, Tokunaga K, and Mizokami M
  • Journal Title: PLoS One Volume: 9 Issue: 2 Pages: e86449-e86449
  • DOI: 10.1371/journal.pone.0086449
  • Peer Reviewed / Open Access
• [Journal Article] A randomized trial of daclatasvir with peginterferon alfa-2b and ribavirin for HCV genotype 1 Infection. (2014)
  • Author(s): Suzuki F, Toyoa J, Ikeda K, Chayama K, Mochida S, Hayashi N, Ishikawa H, Miyagoshi H, Hu W, McPhee F, Hughes
  • Journal Title: Antiviral Ther Volume: in press Issue: 5 Pages: 491-499
  • DOI: 10.3851/imp2730
  • Peer Reviewed
• [Journal Article] The proposal for management of rheumatic disease patients with hepatitis B virus infection receiving immunosuppressive therapy. (2014)
  • Author(s): Harigai M, Mochida S, Mimura T, Koike T, Miyasaka N.
  • Journal Title: Mod Rheumatol Volume: 24 Issue: 1 Pages: 1-7
  • DOI: 10.3109/14397595.2013.852834
  • Peer Reviewed
• [Journal Article] Osteopontin binding to lipopolysaccharide lower tumor necrosis factor-a and prevents early alcohol-induced liver injury in mice. (2014)
  • Author(s): Ge X, Leung TM, Arriazu E, Lu Y, Urtasun R, Christensen B, Fiel MI, Mochida S, Sorensen ES and Nieto N.
  • Journal Title: Hepatology Volume: in press Issue: 4 Pages: 1600-1616
  • DOI: 10.1002/hep.26931
  • Peer Reviewed
• [Journal Article] Telaprevir-Induced, but not Pegylated Interferon-Associated, Retinopathy as a Noteworthy Adverse Effect during Triple Antiviral Therapy in Patients with Chronic Hepatitis C. (2014)
  • Author(s): Sugawara K, Inao M, Nakayama N, Mochida S.
  • Journal Title: J Gastroenterol Volume: 49 Pages: 363-368
  • Peer Reviewed
• [Journal Article] A Possible Novel Genotype HBV Strain Developing Due to Recombination between Genotypes H and B Strains Isolated from a Japanese Patient. (2014)
  • Author(s): Uchida Y, Kouyama J, Naiki K, Sugawara K, Inao M, Nakayama N, Mochida S
  • Journal Title: Hepatol Res Volume: in press Issue: 11 Pages: 1130-1141
  • DOI: 10.1111/hepr.12238
  • Peer Reviewed
• [Journal Article] Therapeutic Strategy for Patients with Bleeding Rectal Varices Complicating Liver Cirrhosis. (2014)
  • Author(s): Yoshino K, Imai Y, Nakazawa M, Chikayama T, Ando S, Sugawara K,Inao M, Oka M, Mochida S.
  • Journal Title: Hepatol Res Volume: in press Issue: 11 Pages: 1088-1094
  • DOI: 10.1111/hepr.12232
  • Peer Reviewed
• [Journal Article] Arterial Chemoembolization for Hepatocellular Carcinoma: Comparison of the Therapeutic Efficacies Between Miriplatin and Epirubicin. (2014)
  • Author(s): Handa T, Imai Y, Sigawara K, Chikayama T, Nakazawa M, Ando S, Hamaorka K, Inao M, Nakayama N, Mochida S.
  • Journal Title: Hepatol Res Volume: in press Issue: 11 Pages: 1072-1080
  • DOI: 10.1111/hepr.12225
  • Peer Reviewed
• [Journal Article] SNPs in the promoter region of osteopontin gene as a possible host factor for sex difference in hepatocellular carcinoma development in patients with HCV. (2013)
  • Author(s): Hamaoka K, Nagoshi S, Sugawara K, Naiki K, Uchida Y, Inao M, Nakayama N, Fujiwara K, Mochida S.
  • Journal Title: Hepatol Int Volume: 7 Issue: 2 Pages: 683-692
  • DOI: 10.1007/s12072-012-9404-1
  • Peer Reviewed
• [Presentation] Frequent Emergences of Rare RAVs Showing Extreme Resistance to NS5A Inhibitors during Dual Oral Therapy with Daclatasvir Plus Asunaprevir in Patients Previously Receiving Triple Therapy with Simeprevir. (2015)
  • Author(s): Yoshihito UCHIDA, Nobuaki NAKAYAMA, Jun-ichi KOUYAMA, Kayoko NAIKI, Hayato UEMURA, Kayoko SUGAWARA, Mie INAO, Satoshi MOCHIDA
  • Organizer: The Liver Meeting, AASLD
  • Place of Presentation: St Francisco,USA
  • Year and Date: 2015-11-13
  • Int'l Joint Research
• [Presentation] Nationwide Prospective Survey in Japan to Clarify the Incidence of Viral Reactivation in Patients with Prior HBV Infection Receiving Immunosuppressive Therapies (2015)
  • Author(s): Masaaki NAKAO, Nobuaki NAKAYAMA, Yoshihito UCHIDA, Masashi MIZOKAMI and Satoshi MOCHIDA
  • Organizer: The Liver Meeting, AASLD
  • Place of Presentation: St Francisco,USA
  • Year and Date: 2015-11-13
  • Int'l Joint Research
• [Presentation] Clinical Features and Outcome of Patients with Acute Liver Failure and LOHF due to HBV Reactivation during and after Immunosuppressive and/or Anticancer Therapies Enrolled in Nationwide Survey in Japan (2015)
  • Author(s): Nobuaki NAKAYAMA, Yoshihito UCHIDA, Masamitsu NAKAO, Hirohito TSUBOUCH, Hjime TAKIKAWA and Satoshi MOCHIDA
  • Organizer: The Liver Meeting, AASLD
  • Place of Presentation: St Francisco,USA
  • Year and Date: 2015-11-13
  • Int'l Joint Research
• [Presentation] NS5A領域のアミノ酸変異に基づいたIFNフリー治療の戦略：サイクリング・プローブ法を用いた検討 (2015)
  • Author(s): 内田義人，菅原通子，持田 智
  • Organizer: JDDW 2015
  • Place of Presentation: 東京 グランドﾌﾟﾘﾝｽホテル新高輪 （東京都港区）
  • Year and Date: 2015-10-08
• [Presentation] NS5Aアミノ酸変異を考慮したC型慢性肝疾患の治療戦略とダクラタスビル・アスナプレビル併用療法の有用性 (2015)
  • Author(s): 中山伸朗，内田義人，持田 智
  • Organizer: 第51回日本肝臓学会総会
  • Place of Presentation: 熊本 ホテル日航熊本（熊本県熊本市）
  • Year and Date: 2015-05-19
• [Presentation] B型肝炎の治療におけるテノホビルの意義 (2015)
  • Author(s): 中尾将光，中山伸朗，持田 智
  • Organizer: 第51回日本肝臓学会総会
  • Place of Presentation: 熊本 ホテル日航熊本（熊本県熊本市）
  • Year and Date: 2015-05-19
• [Presentation] NS5Aアミノ酸変異を有するHCV感染例に対するアスナプレビル・ダクラタスビル経口2剤併用療法の有効性を規定する要因 (2015)
  • Author(s): 内田義人，神山淳一，持田 智
  • Organizer: 第51回日本肝臓学会総会
  • Place of Presentation: 熊本 ホテル日航熊本（熊本県熊本市）
  • Year and Date: 2015-05-19
• [Presentation] The aa15-17 Amino Acid Sequence in the Terminal Protein Domain of the Hepatitis B Virus Polymerase as a Viral Factor Affecting Replication Activity of the Virus (2014)
  • Author(s): Yoshihito UCHIDA, Jun-ichi KOUYAMA, Kayoko NAIKI, MasamitsuNAKAO, Nobuaki NAKAYAMA and Satoshi MOCHIDA
  • Organizer: 第11回Single Topic Conference
  • Place of Presentation: ホテルグランヴィア広島（広島県広島市）
  • Year and Date: 2014-11-20 – 2014-11-21
• [Presentation] HBs抗原量を規定する可能性のある新たなウイルス要因：HBVポリメラーゼ領域 Terminal Protein Domain aa15-17の変異 (2014)
  • Author(s): 内田義人，持田智
  • Organizer: 第100回日本消化器病学会総会
  • Place of Presentation: 東京国際フォーラム（東京都千代田区）
  • Year and Date: 2014-04-23 – 2014-04-26
• [Presentation] The aa15-17 Amino Acid Sequence in the Terminal Protein Domain of HBV Polymerase as a Viral Factor Affecting in Vivo as Well as in Vitro Replication Activity of the Virus
  • Author(s): Uchida Y, Yoshino K, Sugawara K, Kouyama J, Naiki K, Nakayama N, Mochida S
  • Organizer: The liver Meeting 2013 (AASLD)
  • Place of Presentation: Washington,USA