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

2014 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: 自然免疫 / インターフェロン / HCV / HBV

Outline of Annual Research Achievements

末梢血単核球(PBMC)におけるIFN不応性に関わる宿主因子の探索： III型IFNとして新規に発見されたIFNλ4はIL28B SNPのマイナーアレルの症例のみに発現し、これまでの検討では肝内におけるIFNλ4の発現は治療不応性への関与が示唆された。そこで免疫担当細胞であるPBMCにおけるIFN不応性に関わる機序を解明するために、50例のC型慢性肝炎患者のPBMCにおいてIL28BおよびIFNλ4発現とIFN治療応答性の関連について検討した。PBMCを採取後IFNα+poly I:C刺激を行い、無刺激時・刺激時のIFNλ発現、他の自然免疫関連分子発現をRT-qPCRおよびRT-PCRにて解析し、前治療効果(Virological Responder(VR) vs NR)との関連をみた。その結果、無刺激時のIL28B発現レベルはIL28B SNPおよび治療効果との関連を認めなかったが、IFNα+poly I:C刺激によるIL28Bの発現誘導は前治療NR群が再燃例に比して有意にIL28B誘導が低値であり、SNPに関わらずNR群はVR群に比べ有意に低値であった。またMinor群のうち12名でPBMC中にIFNλ4 mRNAの発現が認められ、うち7名では無刺激時のPBMCに発現が認められた。そこで、Minor群のうち、無刺激のPBMC中にIFNλ4発現が検出された群（n=7）と非検出群（n=19）でIL28B発現誘導を比較したところ、IFNλ4検出群で有意にIL28B発現誘導が低値であり（p=0.04）、IFNλ4発現は治療不応性と関連していた。従って、PBMC中のIL28Bの発現誘導能はIFN治療応答性を規定すると考えられ、PBMC に発現するIFNλ4はIFN不応性機序へ関与することが示唆された。

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

PBMCを用いた研究はおおむね順調に進展している。

Strategy for Future Research Activity

新規に発見されたIFNλ4はIL28B SNPのマイナーアレルの症例のみに発現しており、IFN不応性と関連している。しかし、IFN促進系・抑制系に関わるシグナル伝達に与える影響やIL28Bプロモーターに与える影響は不明であり、その受容体も同定されていない。今後さらいIFNλ4が治療不応性に関わる機序について解明し、治療標的としての意義を検討する。また、HBV感染患者ではIFN治療により核内に存在するcccDNAがクロマチン修飾を受け抑制されることが示唆されているが、自然免疫により内因性に誘導されるIFN及びISGがcccDNAに与える影響を解析することで、未だ完全制圧が困難なHBV感染の治療標的としての意義を検討する。

Causes of Carryover

試薬等が計画当初より廉価で購入可能であったため。

Expenditure Plan for Carryover Budget

検討する数・種類を拡大して解析を行うため、試薬を増量して購入する予定である。

Research Products

• [Journal Article] Impaired induction of IL28B and expression of IFNλ4 associated with non-response to interferon-based therapy in chronic hepatitis C. (2015)
  • Author(s): Murakawa M, Asahina Y, Nakagawa M, Sakamoto N, Nitta S, Kusano-Kitazume A, Watanabe T, Kawai-Kitahata F, Otani S, Taniguchi M, Goto F, Nishimura-Sakurai Y, Itsui Y, Azuma S, Kakinuma S, Watanabe M.
  • Journal Title: J Gastroenterol Hepatol Volume: - Pages: -
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Serum granulysin levels as a predictor of serious telaprevir-induced dermatological reactions. (2015)
  • Author(s): Suda G, Yamamoto Y, Nagasaka A, Furuya K, Kudo M, Yoshimichi C, Tsukuda Y, Tsunematsu S, Sato F, Terasita K, Nakai M, Horimoto H, Sho T, Natsuizka M, Ogawa K, Ohnishi S, Chuma M, Fujita Y, Abe R, Taniguchi M, Nakagawa M, Asahina Y, Sakamoto N.
  • Journal Title: Hepatol Res Volume: - Pages: -
  • DOI: 10.1111/hepr.12421
  • Peer Reviewed
• [Journal Article] Hepatocellular carcinoma risk assessment using gadoxetic acid-enhanced hepatocyte phase magnetic resonance imaging. (2015)
  • Author(s): Komatsu N, Motosugi U, Maekawa S, Shindo K, Sakamoto M, Sato M, Tatsumi A, Miura M, Amemiya F, Nakayama Y, Inoue T, Fukasawa M, Uetake T, Ohtaka M, Sato T, Asahina Y, Kurosaki M, Izumi N, Ichikawa T, Araki T, Enomoto N.
  • Journal Title: Hepatol Res Volume: - Pages: -
  • DOI: 10.1111/hepr.12309
  • Peer Reviewed
• [Journal Article] Changes in plasma vascular endothelial growth factor at 8 weeks after sorafenib administration as predictors of survival for advanced hepatocellular carcinoma. (2014)
  • Author(s): Tsuchiya K, Asahina Y, Matsuda S, Muraoak M, Nakata T, Suzuki Y, Tamaki N, Yasui Y, Suzuki S, Hosokawa T, Nishimura T, Ueda K, Kuzuya T, Nakanishi H, Itakura J, Takahashi Y, Kurosaki M, Enomoto N, Izumi N.
  • Journal Title: Cancer Volume: 120 Pages: 229-273
  • DOI: 10.1002/cncr.28384
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Risk factors for exceeding the Milan criteria after successful radiofrequency ablation in patients with early stage hepatocellular carcinoma. (2014)
  • Author(s): Tsuchiya K, Asahina Y, Tamaki N, Yasui Y, Hosokawa T, Ueda K, Nakanishi H, Itakura J, Kurosaki M, Enomoto N, Izumi N.
  • Journal Title: Liver Transpl Volume: 20 Pages: 291-297
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Reduced organic anion transporter expression is a risk factor for hepatocellular carcinoma in chronic hepatitis C patients: A propensity score matching study. (2014)
  • Author(s): Yasui Y, Kudo A, Kurosaki M, Matsuda S, Muraoka M, Tamak Ni, Suzuki S, Hosokawa T, Ueda K, Matsunaga K, Nakanishi H, Tsuchiya K, Itakura J, Takahashi Y, Tanaka S, Asahina Y, Enomoto N, Arii S, Izumi N.
  • Journal Title: Oncology Volume: 86 Pages: 53-62
  • DOI: 10.1159/000356643
  • Peer Reviewed / Acknowledgement Compliant
• [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, Asahina Y, Mizokami M, et al.
  • Journal Title: PLoS One Volume: 9 Pages: e86449
  • DOI: 10.1371/journal.pone.0086449
  • Peer Reviewed
• [Journal Article] Impaired brain activity in cirrhotic patients with minimal hepatic encephalopathy: evaluation by near infrared spectroscopy. (2014)
  • Author(s): Nakanishi H, Kurosaki M, Nakanishi K, Tsuchiya K, Noda T, Tamaki N, Yasui Y, Hosokawa T, Ueda K, Itakura J, Anami K, Asahina Y, Enomoto N, Higuchi T, Izumi N.
  • Journal Title: Hepatol Res Volume: 44 Pages: 319-326
  • DOI: doi: 10.1111/hepr.12127
  • Peer Reviewed
• [Journal Article] Prospective comparison of real-time tissue elastography and serum fibrosis markers for the estimation of liver fibrosis in chronic hepatitis C patients. (2014)
  • Author(s): Tamaki N, Kurosaki M, Matsuda S, Nakata T, Muraoka M, Suzuki Y, Yasui Y, Suzuki S, Hosokawa T, Nishimura T, Ueda K, Tsuchiya K, Nakanishi H, Itakura J, Takahashi Y, Matsunaga K, Taki K, Asahina Y, Izumi N.
  • Journal Title: Hepatol Res Volume: 44 Pages: 720-727
  • DOI: 10.1111/hepr.12179
  • Peer Reviewed
• [Journal Article] JSH guidelines for the management of hepatitis B virus infection. (2014)
  • Author(s): Asahina Y, HayashinN, Hiramatsu N, Izumi N, Koike K, Kumada H, Kurosaki M, Oketani M, Suzuki F, Takikawa H, Tanaka A, Tanaka E, Tanaka Y, Tsubouchi H, Yotsuyanagi H.
  • Journal Title: Hepatol Res Volume: 44 Pages: 1-58
  • DOI: 10.1111/hepr.12269
  • Peer Reviewed
• [Journal Article] JSH guidelines for the management of hepatitis C virus infection: a 2014 update for genotype 1. (2014)
  • Author(s): Asahina Y, HayashinN, Hiramatsu N, Izumi N, Koike K, Kumada H, Kurosaki M, Oketani M, Suzuki F, Takikawa H, Tanaka A, Tanaka E, Tanaka Y, Tsubouchi H, Yotsuyanagi H.
  • Journal Title: Hepatol Res Volume: 44 Pages: 59-70
  • DOI: 10.1111/hepr.12272
  • Peer Reviewed
• [Presentation] Gene Alterations in TERT promoter, CTNNB1, and TP53 are closely associated with development and prognosis of hepatocellular carcinoma: comprehensive analyses by next generation sequencing technology. (2015)
  • Author(s): Asahina Y, Kawai-Kitahata F, Kaneko S, Nagata H, Goto F, Otani S, Taniguchi M, Murakawa M, Nitta S, Watanabe T, Tasaka-Fujita M, Itsui Y, Nakagawa M, Kakinuma S, Tanaka S, Tanabe M, Enomoto N, Watanabe M.
  • Organizer: EASL The International Liver Congress 2015)
  • Place of Presentation: Vienna, Austria
  • Year and Date: 2015-04-23
• [Presentation] Gene alterations in β-catenin and p53/ cell cycle control pathway are closely associated with development and prognosis of hepatocellular carcinoma: Comprehensive analyses by next generation sequencing technology. (2014)
  • Author(s): Kawai-Kitahata F, Asahina Y, Kaneko S, Nagata H, Goto F, Otani S, Taniguchi M, Murakawa M, Nitta S, Watanabe T, Tasaka-Fujita M, Itsui Y, Nakagawa M, Kakinuma S, Enomoto N, Watanabe M.
  • Organizer: The 65th Annual Meeting of the American Association for the Study of Liver Disease (AASLD The Liver Meeting 2014)
  • Place of Presentation: Boston, USA
  • Year and Date: 2014-11-07 – 2014-11-11
• [Presentation] Matrix Metalloproteinase-14 regulates the maturation of fetal hepatic stem/progenitor cells in mice. (2014)
  • Author(s): Otani S, Kakinuma S, Kamiya A, Goto F, Kaneko S, Azuma S, Asahina Y, Watanabe M.
  • Organizer: The 65th Annual Meeting of the American Association for the Study of Liver Disease (AASLD The Liver Meeting 2014)
  • Place of Presentation: Boston, USA
  • Year and Date: 2014-11-07 – 2014-11-11
• [Presentation] Impact of host and therapeutic factors and resistant associated variants on response to interferon based- direct acting antiviral treatment in difficult-to-treat chronic hepatitis C patients. (2014)
  • Author(s): Nakagawa M, Asahina Y, Taniguchi M, Watanabe T, Nishimura-Sakurai Y, Itsui Y, Azuma S, Kakinuma S, Tanaka Y, Watanabe M.
  • Organizer: The 65th Annual Meeting of the American Association for the Study of Liver Disease (AASLD The Liver Meeting 2014)
  • Place of Presentation: Boston, USA
  • Year and Date: 2014-11-07 – 2014-11-11
• [Presentation] Expression of IFN/.4 in liver and PBMC is closely associated with higher basal expression of ISGs and impaired induction of IL28B by interferon treatment in chronic hepatitis C non-responder patients. (2014)
  • Author(s): Murakawa M, Asahina Y, Nakagawa M, Sakamoto N, Nitta S, Kawai-Kitahata F, Taniguchi M, Watanabe T, Itsui Y, Kakinuma S, Watanabe M.
  • Organizer: The 65th Annual Meeting of the American Association for the Study of Liver Disease (AASLD The Liver Meeting 2014)
  • Place of Presentation: Boston, USA
  • Year and Date: 2014-11-07 – 2014-11-11
• [Presentation] Emergence or selection of resistant associated variant immediately after initiation of the therapy is predictive for failure of direct acting antiviral therapy: ultra-deep sequencing analyses for serial time points. (2014)
  • Author(s): Watanabe T, Asahina Y, Nakagawa M, Kakinuma S, YItsui Y, Taniguchi T, Murakawa M, Nagata H, Miura M, Maekawa S, Enomoto E, Watanabe M.
  • Organizer: The 65th Annual Meeting of the American Association for the Study of Liver Disease (AASLD The Liver Meeting 2014)
  • Place of Presentation: Boston, USA
  • Year and Date: 2014-11-07 – 2014-11-11
• [Presentation] Clinical outcomes in patients who develop hepatocellular carcinoma after hepatitis C viral eradication by antiviral therapy. (2014)
  • Author(s): Tsuchiya K, Yasui Y, Tamaki N, Suzuki S, Hosokawa T, Nakanishi H, Itakura J, Takahashi Y, Kurosaki M, Asahina Y, Izumi N.
  • Organizer: The 65th Annual Meeting of the American Association for the Study of Liver Disease (AASLD The Liver Meeting 2014)
  • Place of Presentation: Boston, USA
  • Year and Date: 2014-11-07 – 2014-11-11
• [Presentation] Associations of HLA-DPB1 with CHB infection and HBV related HCC in Asia. (2014)
  • Author(s): Nishida N, Sawai H, Asahina Y, Mizokami M, et al.
  • Organizer: The 65th Annual Meeting of the American Association for the Study of Liver Disease (AASLD The Liver Meeting 2014)
  • Place of Presentation: Boston, USA
  • Year and Date: 2014-11-07 – 2014-11-11
• [Presentation] Impaired IL28B gene induction and expression of IFNλ4 influenced by the polymorphisms near IL28B gene are closely associated with a non-response to interferon in chronic hepatitis C patients. (2014)
  • Author(s): Asahina Y, Murakawa M, NittaS, Itsui Y, Nakagawa M, Azuma S, Kakinuma S, Watanabe M.
  • Organizer: The 49th annual meeting of the European association for the study of the liver (EASL The International Liver Congress 2014)
  • Place of Presentation: London, UK
  • Year and Date: 2014-04-08 – 2014-04-14
• [Presentation] Gadolinium ethoxybenzyl diethylenetriaminepentaacetic acid magnetic resonance imaging and contrast enhanced ultrasonography with sonazoid as part of therapeutic strategies for small nonhypervascular hepatic nodular lesions. (2014)
  • Author(s): Tsuchiya K, Yasui Y, Takada N, Nakakuki S, Matsuda S, Kaneko S, Muraoka M, Yamashita N, Hattori N, Tamaki N, Osaki S, Suzuki T, Hosokawa K, Ueda K, Nakanishi H, Itakura J, Takahashi Y, Kurosaki M, Asahina Y, Izumi N.
  • Organizer: The 49th annual meeting of the European association for the study of the liver (EASL The International Liver Congress 2014)
  • Place of Presentation: London, UK
  • Year and Date: 2014-04-08 – 2014-04-14