自然免疫を基軸としたウイルス性肝炎治療抵抗性機序の解明と新規治療法の創成

2015 Fiscal Year Annual Research Report

• Project/Area Number: 25293169
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: 朝比奈 靖浩 東京医科歯科大学, 医歯(薬)学総合研究科, 寄附講座教授 (00422692)
• Co-Investigator(Kenkyū-buntansha): 柿沼 晴 東京医科歯科大学, 医歯(薬)学総合研究科, 寄附講座講師 (30372444) ; 中川 美奈 東京医科歯科大学, 医歯学融合教育支援センター, 准教授 (30401342)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: HCV / IFNλ3 / IFNλ4 / 自然免疫

Outline of Annual Research Achievements

これまで本研究では、IFN治療不応例では肝内のIFN誘導遺伝子(ISGs)の発現はむしろ高く、III型IFNとして新規に発見されたIFNλ4を肝内に発現している症例では有意に治療抵抗性となることを明らかとしてきた。また、免疫担当細胞である末梢血単核球(PBMC)におけるIFNα+poly I:C刺激時のIFNλ3の発現誘導は治療抵抗例で有意に低値であり、特にIFNλ4が発現している症例ではIFNλ3の発現誘導が有意に低いことから、IFNλ4発現が治療不応性と密接に関連していることが示唆された。そこで、IFNλ4の治療抵抗性に関わる機序を解明するために、IFNλ4（p179）およびその他のsplicing variantからなるIFNλ4強制発現系でのin vitro解析を行った。Huh751におけるp179の強制発現によりISG15が誘導されたが、他のsplicing variant ではISG誘導は見られなかった。一方、p179を強制発現後にIFNα刺激によるISGの誘導能を評価したところ、p179の存在下でISG誘導の減弱が見られた。また、同様の検討をHCVレプリコン細胞で行ったところ、p179の強制発現単独でレプリコン活性の低下が見られたが、IFNα刺激時のHCV減衰はp179存在下の方が弱かった。in vitroにおいて、自然免疫シグナルによるIFNλ3転写活性化能をIFNλ4の存在下・非存在下で検討したところ、p179の共発現によるIFNλ4の存在下ではIRF7が誘導するIFNλ3転写活性の上昇は有意に抑制された。これらのことから、IL28B SNPのリスク・アリルにおいて発現するIFNλ4（p179）はISGを誘導する活性を有するが、外因性のIFNαによるISG誘導及び抗HCV効果を減弱することが、治療不応性と関連すると考えられた。

Research Progress Status

27年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

27年度が最終年度であるため、記入しない。

Causes of Carryover

27年度が最終年度であるため、記入しない。

Expenditure Plan for Carryover Budget

27年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] JSH Guidelines for the Management of Hepatitis C Virus Infection: A 2016 update for genotype 1 and 2. (2016)
  • Author(s): Asahina Y, Izumi N, Hiromitsu K, Kurosaki M, Koike K, Suzuki F, Takikawa H, Tanaka A, Tanaka E, Tanaka Y, Tsubouchi H, Hayashi N, Hiramatsu N, Yotsuyanagi H
  • Journal Title: Hepatol Res Volume: 46 Pages: 129-165
  • DOI: doi: 10.1111/hepr.12645.
  • Peer Reviewed
• [Journal Article] Need for appropriate programs for prevention of hepatitis B infection: lessons from the latest nationwide survey in Japan. (2016)
  • Author(s): Asahina Y, Watanabe M
  • Journal Title: J Gastroenterol Hepatol Volume: 31 Pages: 12-13
  • DOI: doi: 10.1111/jgh.13157
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Evaluation of Interferon Resistance in Newly Established Genotype 1b Hepatitis C Virus Cell Culture System. (2016)
  • Author(s): Taniguchi M, Tasaka-Fujita M, Nakagawa M, Watanabe T, Kawai-Kitahata F, Otani S, Goto F, Nagata H, Kaneko S, Nitta S, Murakawa M, Nishimura-Sakurai Y, Azuma S, Itsui Y, Mori K, Yagi S, Kakinuma S, Asahina Y, Watanabe M
  • Journal Title: J Clin Transl Hepatol Volume: 4 Pages: 5-11
  • DOI: doi: 10.14218/JCTH.2015.00047
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Matrix Metalloproteinase-14 Mediates Formation of Bile Ducts and Hepatic Maturation of Fetal Hepatic Progenitor Cells. (2016)
  • Author(s): Otani S, Kakinuma S, Kamiya A, Goto F, Kaneko S, Miyoshi M, Tsunoda T, Asano Y, Kawai-Kitahata F, Nitta S, Nakata T, Okamoto O, Itsui Y, Nakagawa M, Azuma S, Asahina Y, Yamaguchi T, Koshikawa N, Seiki M, Nakauchi H, Watanabe M
  • Journal Title: Biochem Biophys Res Commun Volume: 469 Pages: 1062-1068
  • DOI: doi: 10.1016/j.bbrc.2015.12.105
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Efficacy of additional radiofrequency ablation after transcatheter arterial chemoembolization for intermediate hepatocellular carcinoma. (2016)
  • Author(s): Azuma S, Asahina Y, Nishimura-Sakurai Y, Kakinuma S, Kaneko S, Nagata H, Goto F, Ootani S, Kawai-Kitahata F, Taniguchi M, Murakawa M, Watanabe T, Tasaka-Fujita M, Itsui Y, Nakagawa M, Watanabe M
  • Journal Title: Hepatol Res Volume: 46 Pages: 312-319
  • DOI: doi: 10.1111/hepr.12566
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Comprehensive analyses of mutations and hepatitis B virus integration in hepatocellular carcinoma with clinicopathological features. (2016)
  • Author(s): Kawai-Kitahata F, Asahina Y, Tanaka S, Kakinuma S, Murakawa M, Nitta S, Watanabe T, Otani S, Taniguchi M, Goto F, Nagata H, Kaneko S, Tasaka-Fujita M, Nishimura-Sakurai Y, Azuma S, Itsui Y, Nakagawa M, Tanabe M , Takano S, Fukasawa M, Sakamoto M, Maekawa S, Enomoto N, Watanabe M
  • Journal Title: J Gastroenterol Volume: 51 Pages: 473-486
  • DOI: doi: 10.1007/s00535-015-1126-4
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Genetic polymorphisms of IL28B and PNPLA3 are predictive for HCV related rapid fibrosis progression and identify patients who require urgent antiviral treatment with new regimens. (2015)
  • Author(s): Tamaki N, Kurosaki M, Higuchi M, Takada T, Nakakuki N, Yasui Y, Suzuki S, Tsuchiya K, Nakanishi H, Itakura J, Takahashi Y, Ogawa S, Tanaka Y, Asahina Y, Izumi N
  • Journal Title: PLoS One Volume: 10 Pages: e0137351
  • DOI: doi: 10.1371/journal.pone.0137351
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Amino acid 1 polymorphisms in hepatitis C virus core affect infectious virus production and major histocompatibility complex class I molecule expression. (2015)
  • Author(s): Tasaka-Fujita M, Sugiyama N, Kang W, Masaski T, Murayama A, Yamada N, Sugiyama R, Tsukuda S, Watashi K, Asahina Y, Sakamoto N, Wakita T, Shin E-C, Kato T
  • Journal Title: Sci Rep Volume: 5 Pages: 13994
  • DOI: doi:10.1038/srep13994
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] Hepatocarcinogenesis after hepatitis C viral eradication. (2016)
  • Author(s): Asahina Y
  • Organizer: The 10th Asian Pacific Association for the Study of the Liver (APASL) Single Topic Conference
  • Place of Presentation: Kaoshiung（Taiwan）
  • Year and Date: 2016-06-08 – 2016-06-10
  • Int'l Joint Research / Invited
• [Presentation] A model for interaction between HBV and host cells derived from human induced pluripotent stem cells. (2016)
  • Author(s): Kaneko S, Kakinuma S, Asahina Y, Kamiya A, Nitta S, Tsunoda T, Miyoshi M, Asano Y, Nagata H, Goto F, Otani S, Kitahata F, Murakawa M, Itsui Y, Nakagawa M, Azuma S, Watanabe M
  • Organizer: The 51th annual meeting of the European association for the study of the liver (EASL The International Liver Congress 2016)
  • Place of Presentation: Barcelona（Spain）
  • Year and Date: 2016-04-13 – 2016-04-18
  • Int'l Joint Research
• [Presentation] Comprehensive analyses of mutations and hepatitis B virus integration in hepatocellular carcinoma. (2016)
  • Author(s): Kawai-Kitahata F, Asahina Y, Tanaka S, Kakinuma S, Murakawa M, Nitta S, Otani S, Goto F, Nagata H, Kaneko S, Asano Y, Miyoshi M, Tsunoda T, Azuma S, Itsui Y, Nakagawa M, Tanabe M, Maekawa S, Enomoto N, Watanabe M
  • Organizer: 25th conference of the Asian Pacific Association for the Study of the Liver (APASL 2016)
  • Place of Presentation: グランドプリンスホテル新高輪（東京都港区）
  • Year and Date: 2016-02-20 – 2016-02-24
  • Int'l Joint Research
• [Presentation] A model for interaction between HBV and host cells derived from human induced pluripotent stem cells. (2016)
  • Author(s): Kaneko S, Kakinuma S, Asahina Y, Kamiya A, Nitta S, Tsunoda T, Miyoshi M, Asano Y, Nagata H, Goto F, Otani S, Kitahata F, Murakawa M, Itsui Y, Nakagawa M, Azuma S, Watanabe M
  • Organizer: 25th conference of the Asian Pacific Association for the Study of the Liver (APASL 2016)
  • Place of Presentation: グランドプリンスホテル新高輪（東京都港区）
  • Year and Date: 2016-02-20 – 2016-02-24
  • Int'l Joint Research
• [Presentation] Serial changes in M2BPGi levels as predictors of fibrosis and HCC in chronic hepatitis C. (2016)
  • Author(s): Nagata H, Sakurai Y, Nakagawa M, Kakinuma S, Azuma S, Murakawa M, Nitta S, Itsui Y, Asahina Y, Watanabe M
  • Organizer: 25th conference of the Asian Pacific Association for the Study of the Liver (APASL 2016)
  • Place of Presentation: グランドプリンスホテル新高輪（東京都港区）
  • Year and Date: 2016-02-20 – 2016-02-24
  • Int'l Joint Research
• [Presentation] In vitro characterization and drug sensitivity analysis for HCV NS5A resistance-associated variants. (2016)
  • Author(s): Nitta S, Asahina Y, Wakita T, Kato T
  • Organizer: 25th conference of the Asian Pacific Association for the Study of the Liver (APASL 2016)
  • Place of Presentation: グランドプリンスホテル新高輪（東京都港区）
  • Year and Date: 2016-02-20 – 2016-02-24
  • Int'l Joint Research
• [Presentation] Human induce pluripotent stem cell-dereived hepatic progenitor-like cells and hepatocyte-like cells as a model for interaction between hepatitis B virus and host cells. (2015)
  • Author(s): Kaneko S, Kakinuma S, Asahina Y, Kamiya A, Nitta S, Tsunoda T, Miyosi M, Nagata H, Goto F, Otani S, Murakawa M, Kawai-Kitahata F, Itsui Y, Nakagawa M, Azuma S, Watanabe M
  • Organizer: The 66th Annual Meeting of the American Association for the Study of Liver Disease (AASLD The Liver Meeting 2015)
  • Place of Presentation: San Francisco（USA）
  • Year and Date: 2015-11-13 – 2015-11-17
  • Int'l Joint Research
• [Presentation] Comprehensive analyses of mutations and hepatitis B virus integration in hepatocellular carcinoma with clinicopathological features. (2015)
  • Author(s): Kawai-Kitahata F, Asahina Y, Tanaka S, Kakinuma S, Murakawa M, Nitta S, Watanabe T, Otani S, Goto F, Nagata H, Kaneko S, Azuma S, Itsui Y, Nakagawa M, Tanabe M, Maekawa S, Enomoto N, Watanabe M
  • Organizer: The 66th Annual Meeting of the American Association for the Study of Liver Disease (AASLD The Liver Meeting 2015)
  • Place of Presentation: San Francisco（USA）
  • Year and Date: 2015-11-13 – 2015-11-17
  • Int'l Joint Research
• [Presentation] Effects of resistance mutations of NS5A inhibitor on viral production and susceptibility to anti-HCV reagents in recombinant hepatitis C viruses with NS5A of genotype 1b. (2015)
  • Author(s): Nitta S, Asahina Y, Wakita T, Kato T
  • Organizer: The 66th Annual Meeting of the American Association for the Study of Liver Disease (AASLD The Liver Meeting 2015)
  • Place of Presentation: San Francisco（USA）
  • Year and Date: 2015-11-13 – 2015-11-17
  • Int'l Joint Research
• [Presentation] Expression of IFN lambda 4 in liver is closely associated with non-response to antiviral therapy through the regulation of basal expression of ISGs in chronic hepatitis C patients but not in hepatitis B. (2015)
  • Author(s): Murakawa M, Asahina Y, Kawai-Kitahata F, Nagata H, Kaneko S, Nitta S, Watanabe T, Itsui Y, Nakagawa M, Kakinuma S, Iijima S, Tanaka Y, Watanabe M, Tanaka Y
  • Organizer: The 66th Annual Meeting of the American Association for the Study of Liver Disease (AASLD The Liver Meeting 2015)
  • Place of Presentation: San Francisco（USA）
  • Year and Date: 2015-11-13 – 2015-11-17
  • Int'l Joint Research
• [Presentation] Serial change of resistant associated variants during early phase of NS3/4A triple therapy and the final virological outcome: analyses by ultra-deep sequenceing technology. (2015)
  • Author(s): Watanabe T, Asahina Y, Nakagawa M, Kakinuma S, Itsui Y, Nagata H, Murakawa M, Kawai-Kitahata F, Miura M, Maekawa S, Enomoto N, Watanabe M
  • Organizer: The 66th Annual Meeting of the American Association for the Study of Liver Disease (AASLD The Liver Meeting 2015)
  • Place of Presentation: San Francisco（USA）
  • Year and Date: 2015-11-13 – 2015-11-17
  • Int'l Joint Research
• [Presentation] Variations of the genome and interaction of hepatitis B viral X protein associated with hepatocarcinogenesis. (2015)
  • Author(s): Nagata H, Itsui Y, Kawai-Kitahata F, Kaneko S, Murakawa M, Nitta S, Nakagawa M, Azuma S, Kakinuma S, Asahina Y
  • Organizer: The 66th Annual Meeting of the American Association for the Study of Liver Disease (AASLD The Liver Meeting 2015)
  • Place of Presentation: San Francisco（USA）
  • Year and Date: 2015-11-13 – 2015-11-17
  • Int'l Joint Research