Metabolic Analysis of the Cultured Hepatic Cell Persistently Infected with Hepatitis C Virus

• Project/Area Number: 23590551
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Virology
• Research Institution: Keio University
• Principal Investigator: SUGIYAMA Kazuo 慶應義塾大学, 医学部, 特任准教授 (10242520)
• Co-Investigator(Renkei-kenkyūsha): SAITO Hidetsugu 慶應義塾大学, 薬学部, 教授 (80186949)
• Project Period (FY): 2011 – 2013
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2013: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2012: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2011: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: C型肝炎ウイルス / ペントースリン酸経路 / コレステロール / 脂肪酸 / Nrf2 / メタボローム / 脂肪滴 / Ｃ型肝炎ウイルス / 持続感染 / ペントースリン酸系 / 代謝 / 脂質代謝 / Hepatitis C virus / hepatocellular carcinoma / metabolome

Research Abstract

In this study, to explore the mechanism for metabolic alteration by hepatisis C virus (HCV), we established a hepatic cell line, HPI cell, persistently infected with HCV. This cell line accumulated massive lipid droplets showing prominent steatosis. Thus we performed integrated analysis by combining microarray and metabolomics. As a result, enhanced production of cholesterol and fatty acids by up-regulation of the rate limiting enzymes. Pentose phosphate pathway was enhanced with production of NADPH. Moreover, TCA cycle was also enhanced indicating hypermetabolic status of the HPI cell. It was speculated that constitutively activated Nrf2, which is a key transcription factor for metabolism, was attributed to the hypermetabolism of HPI cell.
In conclusion, HPI cell is a useful tool for research of not only HCV but also liver metabolism.

Research Products

• [Journal Article] Prominent steatosis with hypermetabolism of the cell line permissive for years of infection with hepatitis C virus (2014)
  • Author(s): Sugiyama, K. Ebinuma, H. Nakamoto, N. Sakasegawa, N. Murakami, Y. Chu, P. S. Usui, S. Ishibashi, Y. Wakayama, Y. Taniki, N. Murata, H. Saito, Y. Fukasawa, M. Saito, K. Yamagishi, Y. Wakita, T. Takaku, H. Hibi, T. Saito, H. Kanai, T.
  • Journal Title: PLoS ONE Volume: 9 Issue: 4 Pages: 849-854
  • DOI: 10.1371/journal.pone.0094460
  • Peer Reviewed / Open Access
• [Journal Article] Free cholesterol accumulation in hepatic stellate cells: Mechanism of liver fibrosis aggravation in nonalcoholic steatohepatitis in mice (2014)
  • Author(s): Tomita, K. Teratani, T. Suzuki, T. Shimizu, M. Sato, H. Narimatsu, K. Okada, Y. Kurihara, C. Irie, R. Yokoyama, H. Shimamura, K. Usui, S. Ebinuma, H. Saito, H. Watanabe, C. Komoto, S. Kawaguchi, A. Nagao, S. Sugiyama, K. Hokari, R. Kanai, T. Miura, S. Hibi, T.
  • Journal Title: Hepatology Volume: 59 Issue: 1 Pages: 154-169
  • DOI: 10.1002/hep.26604
  • Peer Reviewed
• [Journal Article] Hepatitis C Virus Infection Induces Inflammatory Cytokines and Chemokines Mediated by the Cross Talk between Hepatocytes and Stellate Cells. (2013)
  • Author(s): Nishitsuji H, Funami K, Shimizu Y, Ujino S, Sugiyama K, Seya T, Takaku H, Shimotohno K.
  • Journal Title: J. Viol Volume: 87 Issue: 14 Pages: 8169-8178
  • DOI: 10.1128/jvi.00974-13
  • Peer Reviewed
• [Journal Article] Suppressive Effect of the Histone Deacetylase Inhibitor, Suberoylanilide Hydroxamic Acid (SAHA), on Hepatitis C Virus Replication via Epigenetic Changes in Host Cells (2013)
  • Author(s): Sato, A., Saito, Y., Sugiyama, K., Sakasegawa, N., Muramatsu, T., Fukuda, S., Yoneya, M., Kimura, M., Ebinuma, H., Hibi, T., Saito, H
  • Journal Title: J Cell Biochem Volume: 印刷中 Issue: 9 Pages: 1987-1996
  • DOI: 10.1002/jcb.24541
  • Peer Reviewed
• [Journal Article] C-C motif chemokine receptor 9 positive macrophages activate hepatic stellate cells and promote liver fibrosis in mice. (2013)
  • Author(s): Chu PS, Nakamoto N, Ebinuma H, Usui S, Saeki K, Matsumoto A, Mikami Y, Sugiyama K, Tomita K, Kanai T, Saito H, Hibi T.
  • Journal Title: Hepatology Volume: 58 Issue: 1 Pages: 337-350
  • DOI: 10.1002/hep.26351
  • Peer Reviewed
• [Journal Article] p53/p66Shc-mediated signaling contributes to the progression of non-alcoholic steatohepatitis in humans and mice. (2012)
  • Author(s): Tomita K, Teratani T, Shimamura K, Nishiyama N, Mataki R, Irie T, Minamino Y, Okada C, Kurihara H, Sugiyama K, et al
  • Journal Title: J Hepatol Volume: 57 Issue: 4 Pages: 837-843
  • DOI: 10.1016/j.jhep.2012.05.013
  • Peer Reviewed
• [Journal Article] A high-cholesterol diet exacerbates liver fibrosis in mice via accumulation of free cholesterol in hepatic stellate cells (2012)
  • Author(s): Teratani T, Tomita K, Suzuki T, Oshikawa T, Yokoyama H, Shimamura K, Tominaga S, Hiroi S, Irie R, Okada Y, Kurihara C, Ebinuma H, Saito H, Hokari R, Sugiyama K, Kanai T, Miura S, Hibi T
  • Journal Title: Gastroenterology Volume: 142 Issue: 1 Pages: 152-164
  • DOI: 10.1053/j.gastro.2011.09.049
  • Peer Reviewed
• [Journal Article] Development of monoclonal antibodies to human microsomal epoxide hydrolase and analysis of "preneoplastic antigen"-like molecules. (2012)
  • Author(s): Duan H, Yoshimura K, Kobayashi N, Sugiyama K, Sawada JI, Saito Y, Morisseau C, Hammock BD, Akatsuka T
  • Journal Title: Toxicol. Volume: 260 Issue: 1 Pages: 17-26
  • DOI: 10.1016/j.taap.2012.01.023
  • Peer Reviewed
• [Journal Article] Development of monoclonal antibodies to human microsomal epoxide hydrolase and analysis of "preneoplastic antigen"-like molecules. (2012)
  • Author(s): Dua H, Yoshimura K, Kobayashi N, Sugiyama K, Sawada JI, Saito Y, Morisseau C, Hammock BD, Akatsuka T
  • Journal Title: Toxicol. Appl. Pharmacol Volume: 260 Pages: 17-26
  • Peer Reviewed
• [Journal Article] A high-cholesterol diet exacerbates liver fibrosis in mice via accumulation of free cholesterol in hepatic stellate cells. (2012)
  • Author(s): Teratani T, Tomita K, Suzuki T, Oshikawa T, Yokoyama H, Shimamura K, Tominaga S, Hiroi S, Irie R, Okada Y, Kurihara C, Ebinuma H, Saito H, Hokari R, Sugiyama K, Kanai T, Miura S, Hibi T.
  • Journal Title: Gastroenterology Volume: 142
  • Peer Reviewed
• [Journal Article] The interaction between human initiation factor eIF3 subunit c and heat-shock protein 90: a necessary factor for translation mediated by the hepatitis C virus internal ribosome entry site. (2012)
  • Author(s): Ujino S, Nishitsuji H, Sugiyama R, Suzuki H, Hishiki T, Sugiyama K, Shimotohno K, Takaku H
  • Journal Title: Virus Res. Volume: 163 Pages: 390-395
  • Peer Reviewed
• [Journal Article] Lipoprotein component associated with hepatitis C virus is essential for virus infectivity. (2011)
  • Author(s): Shimizu Y, Hishiki T, Ujino S, Sugiyama K, Funami K, Shimotohno K.
  • Journal Title: Current Opinion in Virology Volume: 1 Pages: 19-26
  • Peer Reviewed
• [Presentation] メタボローム・トランスクリプトーム統合的解析によるC型肝炎肝脂肪蓄積の機構解明 (2013)
  • Author(s): 杉山和夫、齋藤英胤,日比紀文
  • Organizer: 第16回日本肝臓学会大会(JDDW 2013)
  • Place of Presentation: 東京
• [Presentation] Epigenetics作用薬を用いたC型肝炎ウイルスの制御 ヒストン脱アセチル化阻害薬の有用性 (2013)
  • Author(s): 齋藤 英胤、齋藤 義正、杉山 和夫
  • Organizer: 第49回日本肝臓学会総会
  • Place of Presentation: 東京
• [Presentation] Nrf2/sMaf転写因子複合体はHCVによる肝細胞脂肪蓄積に関与する (2013)
  • Author(s): 杉山和夫、海老沼浩利、酒瀬川典子、齋藤義正、金井隆典、日比紀文、齋藤英胤
  • Organizer: 第72回日本癌学会総会
  • Place of Presentation: 横浜
• [Presentation] メタボローム・トランスクリプトーム統合的解析によるＣ型肝炎肝脂肪蓄積の機構解明 (2013)
  • Author(s): 杉山和夫
  • Organizer: 第17回日本肝臓学会大会
  • Place of Presentation: 東京
• [Presentation] HCV持続感染細胞における転写因子Nrf2に制御される遺伝子群の発現解析 (2013)
  • Author(s): 酒瀬川典子、杉山和夫、海老沼浩利、中本伸宏、村上優子、齋藤英胤、金井隆典
  • Organizer: 第61回日本ウイルス学会
  • Place of Presentation: 神戸
• [Presentation] Usefulness of A Long-term (Over 2 years) Cultured Cell Clone Persistently and Productively Infected with HCV (2012)
  • Author(s): Sugiyama K, Saito H, Sakasegawa N, Murakami Y, Ebinuma H, Nakamoto N, Usui S, Ishibashi Y, Chu P, Wakayama Y, Saito Y, Kanai T, Hibi T
  • Organizer: The 63rd Annual Meeting of the American Association for the Study of Liver Diseases
  • Place of Presentation: Boston, USA
• [Presentation] New Insights on Hepatic Inflammatory Pseudotumor by Gd-EOB-DTPA MR Imaging (2012)
  • Author(s): Usui S, Chu P, Wakayama Y, Ishibashi Y, Nakamoto N, Yamagishi Y, Masugi Y, Ebinuma H, Sugiyama K, Tanimoto A, Kanai T, Saito H, Hibi T
  • Organizer: ACG 2012 Annual Scientific Meeting
  • Place of Presentation: Las Vegas, USA
• [Presentation] 新たなHCV研究ツールとしてのHCV持続感染培養肝細胞およびその治癒細胞の樹立 (2012)
  • Author(s): 杉山和夫、齋藤英胤、酒瀬川典子、村上優子、海老沼浩利、中本伸宏、梅田瑠美子、碓井真吾、石橋由佳、ちょ柏松、若山遊子、齋藤義正、日比紀文
  • Organizer: 第48回日本肝臓学会総会
  • Place of Presentation: 金沢(ワークショップ)
• [Presentation] HCV研究私の研究とSerendipity、C型肝炎ウイルス持続感染培養肝細胞の樹立と応用 (2012)
  • Author(s): 杉山和夫、齋藤英胤,日比紀文
  • Organizer: 第39回日本肝臓学会東部会
  • Place of Presentation: 東京(シンポジウム)
• [Presentation] Usefulness of a long-term (over 2 years) cultured cell clone persistently and productively infected with chimeric HCV (2012)
  • Author(s): Sugiyama, K, Saito H, Ebinuma H, Sakasegawa N, Murakami Y, Nakamoto N, Usui S, Ishibashi Y, Chu P, Wakayama Y, Saito Y, Kanai T, Hibi T
  • Organizer: The 63rd annual meeting of the American association for the study of liver diseases
  • Place of Presentation: Boston USA
• [Presentation] Recruited CCR9+ macrophages promote murine liver fibrosis through induction of TGF-β in hepatic stellate cells. (2012)
  • Author(s): Chu P, Nakamoto N, Ebinuma H, Usui S, Sugiyama K, Kanai T, Saito H, Hibi T
  • Organizer: The 63rd annual meeting of the American association for the study of liver diseases
  • Place of Presentation: Boston USA
• [Presentation] C型肝炎ウイルス持続感染における肝発癌と免疫異常：高発癌リスク因子を中心とした検討 (2012)
  • Author(s): 楮柏松, 海老沼浩利, 中本伸宏, 碓井真吾, 梅田瑠美子, 石橋由佳, 若山遊子, 菊池真大, 山岸由幸, 杉山和夫, 金井隆典, 齋藤英胤, 日比紀文
  • Organizer: 第４８回日本肝臓学会総会
  • Place of Presentation: 金沢
• [Presentation] 当院における免疫抑制剤投与中のHBV感染の現状 (2012)
  • Author(s): 碓井真吾, 海老沼浩利, 若山遊子, 楮柏松, 石橋由佳, 中本伸宏, 山岸由幸, 杉山和夫, 金井隆典, 齋藤英胤, 日比紀文
  • Organizer: 第４８回日本肝臓学会総会
  • Place of Presentation: 金沢
• [Presentation] コレステロール多量摂取は、肝星細胞の遊離コレステロール蓄積を介して、肝臓繊維化進展を引き起こす (2012)
  • Author(s): 冨田謙吾, 寺谷俊昭, 鈴木貴博, 横山裕一, 岡田義清, 栗原千枝, 海老沼浩利, 齋藤英胤, 渡辺知佳子, 高本俊介, 穂刈量太, 川口淳, 永尾重昭, 杉山和夫, 金井隆典, 日比紀文, 三浦総一郎
  • Organizer: 第４８回日本肝臓学会総会
  • Place of Presentation: 金沢
• [Presentation] ヒストン脱アセチル化酵素阻害剤suberoylanilide hydroxamic acid (SAHA)によるC型肝炎ウイルス(HCV)の増殖抑制効果 (2012)
  • Author(s): 佐藤郁美, 齋藤義正, 永見早耶花, 酒瀬川典子, 福田真也, 木村真規, 杉山和夫, 海老沼浩利, 齋藤英胤
  • Organizer: 第４８回日本肝臓学会総会
  • Place of Presentation: 金沢
• [Presentation] 新たなHCV研究ツールとしてのHCV持続感染培養肝細胞およびその治癒細胞の樹立 (2012)
  • Author(s): 杉山和夫, 齋藤英胤, 酒瀬川典子, 村上優子, 海老沼浩利, 中本伸宏, 梅田瑠美子, 碓井真吾, 石橋由佳, 楮柏松, 若山遊子, 齋藤義正, 日比紀文
  • Organizer: 第４８回日本肝臓学会総会
  • Place of Presentation: 金沢
• [Presentation] Peginterferon (PEG)/ Rivabirin (RBV)併用療法の治療成績からみた高齢者とりわけ高齢女性にたいする今後の治療展望 (2012)
  • Author(s): 海老沼浩利, 齋藤英胤, 中本伸宏, 山岸由幸, 碓井真吾, 梅田瑠美子, 楮柏松, 石橋由佳, 若山遊子, 杉山和夫, 金井隆典, 日比紀文
  • Organizer: 第４８回日本肝臓学会総会
  • Place of Presentation: 金沢
• [Presentation] １ｂ型HCVレプリコン複製細胞内のレプリコンRNAを効率よく感染性粒子としてtranspackagingする方法 (2012)
  • Author(s): 村上優子, 杉山和夫, 齋藤英胤, 海老沼浩利, 中本伸宏, 酒瀬川典子, 斉藤聡, 金井隆典, 日比紀文, 高久洋
  • Organizer: 第６０回日本ウイルス学会学術集会
  • Place of Presentation: 大阪
• [Presentation] 長期間の持続感染培養肝細胞におけるHCVゲノムの自然発生的遺伝子変異の解析 (2012)
  • Author(s): 酒瀬川典子, 杉山和夫, 齋藤英胤, 海老沼浩利, 村上優子, 中本伸宏, 金井隆典, 日比紀文
  • Organizer: 第６０回日本ウイルス学会学術集会
  • Place of Presentation: 大阪
• [Presentation] C型肝炎ウイルス(HCV)複製制御に係わる宿主因子Hsp70の機能解析 (2012)
  • Author(s): 小倉大直, 君塚圭亮, 杉山隆一, 日紫喜隆行, 下遠野邦忠, 杉山和夫, 高久洋
  • Organizer: 第６０回日本ウイルス学会学術集会
  • Place of Presentation: 大阪
• [Presentation] Age and the liver stiffness measured by Fibroscan are significant predictive values for hepatoma development in chronic liver diseases (2011)
  • Author(s): Ishibashi Y, Ebinuma H, Sugiyama K, Nakamoto N, Yamagishi Y, Usui S, Cho HS, Umeda R, Wakayama Y, Takaishi H, Kanai T, Sakamoto M, Saito H, Hibi T
  • Organizer: 62nd Annual Meeting of the American Association for the Study of Liver Diseases
  • Place of Presentation: San Francisco, USA
• [Presentation] Significant and continuous elevation of serum total cholesterol levels in patients with chronic hepatitis C who achieved sustained virological response by peginterferon and rivabirin treatment (2011)
  • Author(s): Ebinuma H, Saito H, Sugiyama K, Nakamoto N, Yamagishi Y, Usui S, Cho HS, Umeda R, Ishibashi Y, Wakayama Y, Kanai T, Hibi T
  • Organizer: 62nd Annual Meeting of the American Association for the Study of Liver Diseases
  • Place of Presentation: San Francisco, USA
• [Presentation] キメラHCV(1b/2a)持続感染細胞のメタボローム解析(Metabolome analysis of the cultured hepatocytes persistently infected with Chimeric HCV (1b/2a) (2011)
  • Author(s): 杉山和夫、齋藤英胤、海老沼浩利、高久洋、日比紀文
  • Organizer: 第70回日本癌学会総会
  • Place of Presentation: 名古屋
• [Presentation] Nrf2/sMaf転写因子複合体はHCVによる肝細胞脂肪蓄積に関与する (2011)
  • Author(s): 杉山和夫、海老沼浩利、酒瀬川典子、齋藤義正、金井隆典、日比紀文、齋藤英胤
  • Organizer: 第70回日本癌学会総会
  • Place of Presentation: 名古屋
• [Presentation] Infectious hepatitis C virus bearing genotype-1b subgenomic replicon by trans-encapsidation with the structural regions of genotype 1b. (2011)
  • Author(s): Sugiyama, K, Murakami Y, Hishiki T, Shimizu Y, Funami K, Ujino S, Saito H, Saito S, Kato N, Hibi T, Takaku H
  • Organizer: The 18th International Symposium on Hepatitis C Virus and Related Viruses
  • Place of Presentation: Seattle USA
• [Presentation] Metabolome analysis of the cultured hepatocytes persistently infected with Chimeric HCV (1b/2a). (2011)
  • Author(s): Sugiyama, K, Saito H, Ebinuma H, Takaku H, Hibi T
  • Organizer: 第70回日本癌学会総会
  • Place of Presentation: 名古屋
• [Presentation] Metabolome analysis of the cultured hepatocytes persistently infected with Chimeric HCV (1b/2a). (2011)
  • Author(s): Sugiyama, K. , Saito, H., Ebinuma, H., Takaku, H., Hibi, T.
  • Organizer: 70th Annual Meeting of the Japanese Cancer Association.
  • Place of Presentation: 名古屋国際会議場(愛知県）
• [Presentation] Infectious hepatitis C virus bearing genotype-1b subgenomic replicon by trans-encapsidation with the structural regions of genotype 1b. (2011)
  • Author(s): Sugiyama, K., Murakami, Y., Hishiki, T., Shimizu, Y., Funami, K., Ujino, S., Saito, H., Saito, S., Kato, N., Hibi, T., Takaku, H.
  • Organizer: 18th International Symposium on Hepatitis C Virus and Related Viruses
  • Place of Presentation: シアトル、アメリカ合衆国
• [Book] がん生物学イラストレイティド, 腫瘍ウイルス(肝炎ウイルス) (2011)
  • Author(s): 杉山和夫, (渋谷正史,湯浅保仁編)
  • Publisher: 羊土社
• [Book] がん生物学イラストレイテッド、腫瘍ウイルス(肝炎ウイルス) (2011)
  • Author(s): 宇治野真之, 杉山和夫, 下遠野邦忠
  • Total Pages: 10
  • Publisher: 羊土社