The role of PKR in hepatocellular carcinoma associated with intracellular metabolism

• Project/Area Number: 20K08388
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 53010:Gastroenterology-related
• Research Institution: Ehime University
• Principal Investigator: Watanabe Takao 愛媛大学, 医学系研究科, 講師 (90650458)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2022: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2021: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
• Keywords: 肝細胞癌 / PKR / 細胞増殖 / 代謝経路 / 阻害剤 / 代謝 / 解糖系 / 結合蛋白質 / NASH / 肝線維化

Outline of Research at the Start

Protein kinase R (PKR)は一部の代謝関連蛋白質に影響を与えており、NASH関連肝細胞癌における、PKRの代謝異常を介した役割を想定した。PKRノックアウトマウスを作製し、二光子励起イメージング技術により、PKRが肝線維化および肝細胞癌発症に与える影響を明らかにする。またメタボロミクス, ゲノミクス, プロテオミクス解析を駆使し、PKRが制御している代謝酵素のスクリーニングを行う。得られた候補分子については生体イメージングシステムの応用により機能解析を行う。以上の結果、NASH関連肝細胞癌の分子メカニズム・治療標的を代謝異常という新しい観点から明らかにし、創薬に結びつける。

Outline of Final Research Achievements

We have shown that PKR is highly expressed in hepatocellular carcinoma and that administration of imidazolo-oxindole, an inhibitor of PKR suppressed tumor growth. We also identified by mass spectrometry hexokinase -2, a major enzyme in glycolysis, as a target protein for PKR inhibitor. In hepatocellular carcinoma cell lines PKR inhibitor treatment suppressed HK2 expression. We showed that PKR and HK2 bind directly, which is dependent on PKR phosphorylation. PKR inhibitors suppressed intracellular ATP production by suppressing glycolysis. Overexpression of HK2 increased tumor proliferation and adding overexpression of PKR at the same time additively increased that. These findings suggest that PKR-HK2 axis increases tumor growth in hepatocellular carcinoma by promoting glycolysis, and that PKR-HK2 axis may be a novel therapeutic target in hepatocellular carcinoma.

Academic Significance and Societal Importance of the Research Achievements

肝細胞癌における治療標的となる分子や経路はごく限られており、治療効果のエビデンスのある薬物は十分ではないのが現状である。我々は以前より肝細胞癌で高発現するPKRの役割、癌促進機序を研究してきた。本研究ではPKRが肝細胞癌の細胞内代謝に関わる複数の分子に影響を与えていることを見出した。その中で特に解糖系の主要分子であるHK2に着目し研究を行い、PKR-HK2経路が解糖系の促進を介して腫瘍増殖能を亢進していることを見出した。このことは細胞内代謝経路の変化という新たな視点での治療標的になり得ると考えられた。

Research Products

• [Journal Article] Role of B Cell-Activating Factor in Fibrosis Progression in a Murine Model of Non-Alcoholic Steatohepatitis (2023)
  • Author(s): Kozue Kanemitsu-Okada, Masanori Abe, Yoshiko Nakamura, Teruki Miyake, Takao Watanabe, Osamu Yoshida, Yohei Koizumi, Masashi Hirooka, Yoshio Tokumoto, Bunzo Matsuura, Mitsuhito Koizumi, Yoichi Hiasa
  • Journal Title: International Journal of Molecular Sciences Volume: 24 Issue: 3 Pages: 2509-2509
  • DOI: 10.3390/ijms24032509
  • Peer Reviewed / Open Access
• [Journal Article] Spleen stiffness in patients with chronic liver disease evaluated by 2-D shear wave elastography with ultrasound multiparametric imaging (2023)
  • Author(s): Hirooka M, Koizumi Y, Nakamura Y, Yano R, Okazaki Y, Sunago K, Imai Y, Watanabe T, Yoshida O, Tokumoto Y, Abe M, Hiasa Y.
  • Journal Title: Hepatology Research Volume: 53 Issue: 2 Pages: 93-103
  • DOI: 10.1111/hepr.13841
  • Peer Reviewed
• [Journal Article] Cisterna chyli as an optimal marker of tolvaptan response in severe cirrhotic ascites (2022)
  • Author(s): Hirooka M, Koizumi Y, Yano R, Nakamura Y, Sunago K, Yukimoto A, Watanabe T, Yoshida O, Tokumoto Y, Abe M, Hiasa Y.
  • Journal Title: Scientic Reports Volume: 17 Issue: 1 Pages: 8124-8124
  • DOI: 10.1038/s41598-022-11889-z
  • Peer Reviewed / Open Access
• [Journal Article] Plasma Fatty Acid Composition Is Associated with Histological Findings of Nonalcoholic Steatohepatitis (2022)
  • Author(s): Teruki Miyake, Shinya Furukawa, Bunzo Matsuura, Osamu Yoshida, Masumi Miyazaki, Akihito Shiomi, Sayaka Kanzaki, Hironobu Nakaguchi, Kotaro Sunago, Yoshiko Nakamura, Yusuke Imai, Takao Watanabe, Yasunori Yamamoto, Yohei Koizumi, Yoshio Tokumoto, Masashi Hirooka, Teru Kumagi, Masanori Abe, Yoichi Hiasa
  • Journal Title: Biomedicines Volume: 10 Issue: 10 Pages: 2540-2540
  • DOI: 10.3390/biomedicines10102540
  • Peer Reviewed / Open Access
• [Journal Article] Additional Effect of Luseogliflozin on Semaglutide in Nonalcoholic Steatohepatitis Complicated by Type 2 Diabetes Mellitus: An Open-Label, Randomized, Parallel-Group Study (2022)
  • Author(s): Miyake T, Yoshida O, Matsuura B, Furukawa S, Hirooka M, Abe M, Tokumoto Y, Koizumi Y, Watanabe T, Takeshita E, Sunago K, Yukimoto A, Watanabe K, Miyazaki M, Kanzaki S, Nakaguchi H, Koizumu M, Yamamoto Y, Kumagi T, Hiasa Y.
  • Journal Title: Diabetes Therapy Volume: 13 Issue: 5 Pages: 1083-1096
  • DOI: 10.1007/s13300-022-01239-7
  • Peer Reviewed / Open Access
• [Journal Article] AFP and eGFR are related to early and late recurrence of HCC following antiviral therapy. (2021)
  • Author(s): Watanabe T, Tokumoto Y, Joko K, Michitaka K, Horiike N, Tanaka Y, Tada F, Kisaka Y, Nakanishi S, Yamauchi K, Ochi H, Hiraoka A, Yagi S, Yukimoto A, Hirooka M, Abe M, Hiasa Y
  • Journal Title: BMC Cancer Volume: 14;21(1) Issue: 1 Pages: 699-699
  • DOI: 10.1186/s12885-021-08401-7
  • Peer Reviewed / Open Access
• [Journal Article] The long noncoding RNA of RMRP is downregulated by PERK, which induces apoptosis in hepatocellular carcinoma cells (2021)
  • Author(s): Yukimoto Atsushi、Watanabe Takao、Sunago Kotaro、Nakamura Yoshiko、Tanaka Takaaki、Koizumi Yohei、Yoshida Osamu、Tokumoto Yoshio、Hirooka Masashi、Abe Masanori、Hiasa Yoichi
  • Journal Title: Scientific Reports Volume: 11 Issue: 1 Pages: 7926-7926
  • DOI: 10.1038/s41598-021-86592-6
  • Peer Reviewed / Open Access
• [Journal Article] Apoptosis-associated speck-like protein containing a CARD regulates the growth of pancreatic ductal adenocarcinoma (2021)
  • Author(s): Koizumi Mitsuhito、Watanabe Takao、Masumoto Junya、Sunago Kotaro、Imamura Yoshiki、Kanemitsu Kozue、Kumagi Teru、Hiasa Yoichi
  • Journal Title: Scientific Reports Volume: 11 Issue: 1 Pages: 22351-22351
  • DOI: 10.1038/s41598-021-01465-2
  • Peer Reviewed / Open Access
• [Journal Article] Validation of the FibroScan-aspartate aminotransferase score by vibration-controlled transient and B-mode ultrasound elastography. (2021)
  • Author(s): Hirooka M, Koizumi Y, Yano R, Sunago K, Watanabe T, Yoshida O, Tokumoto Y, Abe M, Hiasa Y.
  • Journal Title: Hepatology Research Volume: 51(6) Issue: 6 Pages: 652-661
  • DOI: 10.1111/hepr.13646
  • Peer Reviewed
• [Journal Article] Relationship between body composition and the histology of non‐alcoholic fatty liver disease: a cross‐sectional study (2021)
  • Author(s): Miyake Teruki、Miyazaki Masumi、Yoshida Osamu、Kanzaki Sayaka、Nakaguchi Hironobu、Nakamura Yoshiko、Watanabe Takao、Yamamoto Yasunori、Koizumi Yohei、Tokumoto Yoshio、Hirooka Masashi、Furukawa Shinya、Takeshita Eiji、Kumagi Teru、Ikeda Yoshio、Abe Masanori、Toshimitsu Kumiko、Matsuura Bunzo、Hiasa Yoichi
  • Journal Title: BMC Gastroenterology Volume: 21 Issue: 1 Pages: 170-170
  • DOI: 10.1186/s12876-021-01748-y
  • Peer Reviewed / Open Access
• [Journal Article] Direct-acting antivirals improve survival and recurrence rates after treatment of hepatocellular carcinoma within the Milan criteria (2021)
  • Author(s): Ochi H, Hiraoka A, Hirooka M, Koizumi Y, Amano M, Azemoto N, Watanabe T, Yoshida O, Tokumoto Y, Mashiba T, Yokota T, Abe M, Michitaka K, Hiasa Y, Joko K.
  • Journal Title: Journal of Gastroenterology Volume: 56 Issue: 1 Pages: 90-100
  • DOI: 10.1007/s00535-020-01747-y
  • Peer Reviewed / Open Access
• [Journal Article] Sex difference in the development of hepatocellular carcinoma after direct-acting antiviral therapy in patients with HCV infection. J Med Virol. (2020)
  • Author(s): Watanabe T, Tokumoto Y, Joko K, Michitaka K, Horiike N, Tanaka Y, Tada F, Kisaka Y, Nakanishi S, Yamauchi K, Yukimoto A, Nakamura Y, Hirooka M, Abe M, Hiasa Y
  • Journal Title: Journal of Medical Virology Volume: 92 Issue: 12 Pages: 3507-3515
  • DOI: 10.1002/jmv.25984
  • Peer Reviewed
• [Journal Article] Advanced fibrosis of non-alcoholic steatohepatitis affects the significance of lipoprotein(a) as a cardiovascular risk factor (2020)
  • Author(s): Konishi K, Miyake T, Furukawa S, Senba H, Kanzaki S, Nakaguchi H, Yukimoto A, Nakamura Y, Watanabe T, Koizumi Y, Yoshida O, Tokumoto Y, Hirooka M, Kumagi T, Abe M, Matsuura B, Hiasa Y.
  • Journal Title: Atherosclerosis Volume: 299 Pages: 32-37
  • DOI: 10.1016/j.atherosclerosis.2020.02.026
  • Peer Reviewed
• [Presentation] SVR後の食道胃静脈瘤増悪に寄与する因子の検討 (2022)
  • Author(s): 渡辺崇夫、徳本良雄、日浅陽一
  • Organizer: 第108回日本消化器病学会総会
• [Presentation] SVR後の食道胃静脈瘤増悪・改善を予測可能なスコアリングシステムの作成 (2022)
  • Author(s): 渡辺崇夫、徳本良雄、日浅陽一
  • Organizer: 第58回日本肝臓学会総会
• [Presentation] 年齢層に応じたSVR後肝発癌危険因子の検討 (2022)
  • Author(s): 渡辺崇夫、徳本良雄、日浅陽一
  • Organizer: 第26回日本肝臓学会大会
• [Presentation] A simple and accurate prediction model of hepatocellular carcinoma occurrence after DAA therapy using the factors at SVR12 (2022)
  • Author(s): Takao Watanabe, Yoshio Tokumoto, Makoto Morita, Ryo Yano, Yuki Okazaki, Kotaro Sunago, Yoshiko Nakamura, Yusuke Imai, Yohei Koizumi, Osamu Yoshida, Masashi Hirooka, Masanori Abe, and Yoichi Hiasa
  • Organizer: AASLD the liver meeting 2022
  • Int'l Joint Research
• [Presentation] SVR後の食道胃静脈瘤増悪を予測する因子の検討 (2021)
  • Author(s): 渡辺崇夫、徳本良雄、上甲康二、道堯浩二郎、堀池典生、田中良憲、木阪吉保、多田藤政、中西征司、八木専、山内一彦、廣岡昌史、阿部雅則、日浅陽一
  • Organizer: 第56回日本肝臓学会総会
• [Presentation] SVR後の肝癌サーベイランス法の検証 (2021)
  • Author(s): 渡辺崇夫、徳本良雄、日浅陽一
  • Organizer: 第24回日本肝臓学会大会
• [Presentation] 小胞体ストレスに関連する long noncoding RNA RMRP を介した肝細胞がんのアポトーシス誘導作用 (2021)
  • Author(s): 行本 敦，渡辺崇夫，日浅陽一
  • Organizer: 第44回日本肝臓学会西部会
• [Presentation] Predictors of hepatocellular carcinoma recurrence after direct-acting antiviral therapy in patients with HCV infection (2021)
  • Author(s): Takao Watanabe, Yoshio Tokumoto, Atsushi Yukimoto, Yuki Okazaki, Kotaro Sunago, Yohei Koizumi, Osamu Yoshida, Masashi Hirooka, Masanori Abe, Yoichi Hiasa
  • Organizer: JSH International Liver Conference 2021
  • Int'l Joint Research
• [Presentation] Sex-related Differences in Predictors of HCC Incidence after DAA Therapy in Patients with HCV (2021)
  • Author(s): Takao Watanabe, Yoshio Tokumoto, Atsushi Yukimoto, Yoshiko Nakamura, Yuki Okazaki, Kotaro Sunago, Yohei Koizumi, Osamu Yoshida, Masashi Hirooka, Masanori Abe, Yoichi Hiasa
  • Organizer: APASL Oncology 2021
  • Int'l Joint Research
• [Presentation] 肝細胞癌におけるProtein kinase R (PKR)の組織バイオマーカーとしての役割と標的治療の可能性 (2020)
  • Author(s): 渡辺崇夫、徳本良雄、日浅陽一
  • Organizer: 第55回日本肝臓学会総会
• [Presentation] 肝細胞癌におけるprotein kinase R (PKR)の治療標的としての可能性 (2020)
  • Author(s): 渡辺崇夫、二宮寛子、齋藤卓、川村良介、小泉光仁、吉田理、徳本良雄、廣岡昌史、阿部雅則、今村健志、日浅陽一
  • Organizer: 第79回日本癌学会学術集会
• [Presentation] 肝細胞癌における Protein kinase R (PKR)の役割と、治療標的としての可能性 (2020)
  • Author(s): 渡辺崇夫、今村健志、日浅陽一
  • Organizer: 第24回日本がん分子標的治療学会学術集会
• [Presentation] SOF/VEL治療における肝予備能評価指数の変化と改善を予測する因子の検討 (2020)
  • Author(s): 渡辺崇夫、真柴寿枝、上甲康二、道堯浩二郎、徳本良雄、日浅陽一
  • Organizer: 第24回日本肝臓学会大会
• [Presentation] DAA治療後の時期に応じた肝細胞癌再発寄与因子の検討 (2020)
  • Author(s): 渡辺崇夫、上甲康二、道堯浩二郎、堀池典生、田中良憲、多田藤政、木阪吉保、中西征司、山内一彦、徳本良雄、日浅陽一
  • Organizer: 第62回日本消化器病学会大会