Mandatory autophagy induction targeting gastric cancer stem cells by hybrid small molecule antibodies

2018 Fiscal Year Annual Research Report

• Project/Area Number: 16H05291
• Research Institution: Keio University
• Principal Investigator: 鈴木 秀和 慶應義塾大学, 医学部(信濃町), 教授 (70255454)
• Co-Investigator(Kenkyū-buntansha): 津川 仁 慶應義塾大学, 医学部(信濃町), 講師 (30468483) ; 土居 信英 慶應義塾大学, 理工学部(矢上), 教授 (50327673)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Keywords: オートファジー / CD44v9 / 癌幹細胞 / H. pylori / CagA / ペプチド

Outline of Annual Research Achievements

H. pylori感染細胞におけるオートファジー発現は本菌由来がん蛋白質CagA分解を担うため、癌化抑制に寄与する重要な宿主応答である。しかし、CD44v9発現癌幹細胞では、このオートファジーが発現せず細胞内CagAが分解されずに蓄積するため、胃癌発生・再発に関連する癌幹細胞となると考えられた。CagAのオートファジー分解はautophagosome形成だけでなくLAMP1依存性autophagolysosome形成にも必須であり、我々はアクチン重合制御因子CAPZA1が核内でLRP1-ICDと結合しLAMP1発現を阻害することを明らかにし、かつ、オートファジー発現不全のCAPZA1過剰発現細胞にCagAが装填されるとCagAは分解されずにCD44v9発現が亢進することを示し、CD44v9発現癌幹細胞を標的するためにCD44v9発現の標的に加え核内移行した大量のCAPZA1を標的する必要があると考えた。そこで、CD44v9を標的しCD44v9陽性細胞特異的自殺因子を細胞質内送達する目的で、ヒトSyncytin1由来膜透過促進ペプチドS19とTAT とを組み合わせたところ、細胞質内送達効率を数十倍に向上しえた。さらに初期エンドソーム（EE）と後期エンドソーム（LE）の脂質膜組成模倣リポソームを調製しS19-TATペプチドを作用させたところ、S19-TATはEEではなくLEから細胞質に離脱し、S19-TATは、細胞内異物分解機構から離脱可能であることが示された。次に、CD44v9陽性細胞に標的物質を特異的送達するためmRNAディスプレイ法で取得した抗CD44v9一本鎖抗体とヒト由来膜透過促進ペプチドを融合した蛍光タンパクを大量精製しCD44v9陽性細胞選択的な細胞内取り込みを確認した。以上から、CD44v9発現細胞標的療法の基盤が構築された。

Research Progress Status

平成30年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

平成30年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] CAPZA1 determines the risk of gastric carcinogenesis by inhibiting Helicobacter pylori CagA-degraded autophagy (2019)
  • Author(s): Tsugawa H, Mori H, Matsuzaki J, Sato A, Saito Y, Imoto M, Suematsu M, Suzuki H
  • Journal Title: Autophagy Volume: 15 Pages: 242-258
  • DOI: 10.1080/15548627.2018.1515530
  • Peer Reviewed
• [Journal Article] Clinical and endoscopic findings to assist the early detection of duodenal adenoma and adenocarcinoma. (2019)
  • Author(s): Matsuzaki, J., Suzuki, H.*, Shimoda, M., Mori, H., Fukuhara, S., Miyoshi, S., Masaoka, T., Iwao, Y., Kanai, T.
  • Journal Title: United European Gastroenterology Journal Volume: 7 Pages: 250-260
  • DOI: https://doi.org/10.1177/2050640618817689
  • Peer Reviewed
• [Journal Article] Burden of impaired sleep quality on work productivity in functional dyspepsia (2018)
  • Author(s): Matsuzaki, J., Suzuki, H.*, Togawa, K., Yamane, T., Mori, H., Komori, T., Masaoka, T., Kanai, T. .
  • Journal Title: United European Gastroenterology Journal Volume: 6 Pages: 398-406
  • DOI: https://doi.org/10.1177/2050640617726466.
  • Peer Reviewed
• [Journal Article] Pethidine dose and female sex as risk factors for nausea after esophagogastroduodenoscopy. (2018)
  • Author(s): Nishizawa, T., Suzuki, H., Arita, M., Kataoka, Y., Fukagawa, K., Ohki, D., Hata, K., Uraoka, T., Kanai, T., Yahagi, N., Toyoshima, O.
  • Journal Title: J Clin Biochem Nutr. Volume: 63 Pages: 230-232
  • DOI: 10.3164/jcbn.18-5.
  • Peer Reviewed
• [Journal Article] Autotaxin and vascular endothelial growth factor receptor-2 and -3 are related to vascular development during the progression of chronic viral hepatitis C. (2018)
  • Author(s): Yokomori, H., Ando, W., Kaneko, F., Suzuki, H., Igarashi, K., Oda, M.
  • Journal Title: APMIS Volume: 126 Pages: 913-921
  • DOI: 10.1111/apm.12904
  • Peer Reviewed
• [Journal Article] Inhibiting xCT improves 5-fluorouracil resistance of gastric cancer induced by CD44 variant 9 expression. (2018)
  • Author(s): Miyoshi, S., Tsugawa, H., Matsuzaki, J., Hirata, K., Mori, H., Saya, H., Kanai, T., Suzuki, H.*
  • Journal Title: Anticancer Res. Volume: 38 Pages: 6163-6170
  • DOI: 10.21873/anticanres.12969.
  • Peer Reviewed
• [Journal Article] Pathophysiology of potassium-competitive acid blocker-refractory gastroesophageal reflux and the potential of potassium-competitive acid blocker test. (2018)
  • Author(s): Masaoka, T., Kameyama, H., Yamane, T., Yamamoto, Y., Takeuchi, H., Suzuki, H., Kitagawa, Y., Kanai, T.
  • Journal Title: J Neurogastroenterol Motil. Volume: 24 Pages: 577-583
  • DOI: 10.5056/jnm18036
  • Peer Reviewed
• [Journal Article] Serum anti-Helicobacter pylori antibody titer and its association with gastric nodularity,atrophy,and age: A cross-sectional study. (2018)
  • Author(s): Toyoshima, O., Nishizawa, T., Sakitani, K., Yamakawa, T., Takahashi, Y., Yamauchi, N., Hata, K., Seto, Y., Koike, K., Watanabe, H., Suzuki, H.
  • Journal Title: World J. Gastroenterol. Volume: 24 Pages: 4061-4068
  • DOI: 10.3748/wjg.v24.i35.4061
  • Peer Reviewed
• [Journal Article] Serum autotaxin concentrations reflect changes in liver stiffness and fibrosis after antiviral therapy in patients with chronic hepatitis C. (2018)
  • Author(s): Ando, W., Yokomori, H., Kaneko, F., Kaneko, M., Igarashi, K., Suzuki, H.*
  • Journal Title: Hepatology Communications Volume: 2 Pages: 1111-1122
  • DOI: 10.1002/hep4.1230.
  • Peer Reviewed
• [Journal Article] Helicobacter pylori infection in subjects negative for high titer serum antibody. (2018)
  • Author(s): Toyoshima, O., Nishizawa, T., Arita, M., Kataoka, Y., Sakitani, K., Yoshida, S., Yamashita, H., Hata, K., Watanabe, H., Suzuki, H.
  • Journal Title: World J. Gastroenterol. Volume: 24 Pages: 1419-1428
  • DOI: 10.3748/wjg.v24.i13.1419.
  • Peer Reviewed
• [Journal Article] Acquisition of double mutation in gyrA caused high resistance to sitafloxacin in Helicobacter pylori after unsuccessful eradication with sitafloxacin-containing regimens. (2018)
  • Author(s): Mori, H., Suzuki, H.*, Matsuzaki, J., Masaoka, T., Kanai, T.
  • Journal Title: United European Gastroenterology Journal Volume: 6 Pages: 391-397
  • DOI: https://doi.org/10.1177/2050640617737215.
  • Peer Reviewed
• [Journal Article] Plausibility criteria for putative pathophysiological mechanisms in functional gastrointestinal disorders: a consensus of experts. (2018)
  • Author(s): Tack, J., Corsetti, M., Camilleri, Mi., Quigley, A., Simren, M., Suzuki, H., Talley, N.J., Tornbloom, H., Van Oudenhove, L.
  • Journal Title: Gut Volume: 67 Pages: 1425-1433
  • DOI: 10.1136/gutjnl-2016-312230.
  • Peer Reviewed / Int'l Joint Research
• [Presentation] H. pylori eradication: past, present and future. (2018)
  • Author(s): Suzuki, H.
  • Organizer: APDW2018, Seoul, Korea
  • Int'l Joint Research / Invited
• [Presentation] Nationwide eradication strategy for Helicobacter pylori infection in Japan. (2018)
  • Author(s): Suzuki, H.
  • Organizer: APDW2018, Seoul, Korea
  • Int'l Joint Research / Invited
• [Presentation] Gender difference in gastrointestinal function and functional GI disorders. (2018)
  • Author(s): Suzuki, H.
  • Organizer: APDW2018, Seoul, Korea
  • Int'l Joint Research / Invited
• [Presentation] The evolving landscape of H. pylori, Relation to gastric cancer. Sponsored seminar lecture. (2018)
  • Author(s): Suzuki, H.
  • Organizer: The International Sentinel Node Society Biennial Meeting 2018 (ISNS 2018) Tokyo, Japan
  • Int'l Joint Research / Invited
• [Presentation] Case Presentation and Q&A - Novel approaches for PPI refractory GERD. Plenary Session 11: Novel approaches for PPI refractory GERD. (2018)
  • Author(s): Suzuki, H.
  • Organizer: The International Society for Diseases of the Esophagus (ISDE) 2018. VIenna, Austria
  • Int'l Joint Research / Invited
• [Presentation] Refux hypersensitivity: a new entity? (2018)
  • Author(s): Suzuki, H.
  • Organizer: GUT 2018 (Annual Scientific Meeting of the Malaysian Society of Gastroenterology & Hepatology) Penang, Malaysia
  • Invited
• [Presentation] The evolving landscape of Helicobacter pylori in Asia. (2018)
  • Author(s): Suzuki, H.
  • Organizer: GUT 2018 (Annual Scientific Meeting of the Malaysian Society of Gastroenterology & Hepatology) Penang, Malaysia
  • Invited
• [Presentation] Recent Advance in the therapeutic strategy for acid-related diseases. (2018)
  • Author(s): Suzuki, H.
  • Organizer: 58th GAT (Gastrointestinal Association of Thailand) Midyear Annual Meeting 2018, Bangkok, Thailand
  • Invited
• [Presentation] Association between oxidative stress and gastric carcinogenesis in H. pylori-infected patients. (2018)
  • Author(s): Tsugawa, H., Kato, C., Mori, H., Suematsu, M., Suzuki, H.
  • Organizer: Summer School on Stress 2018, Osaka, Japan
  • Int'l Joint Research
• [Presentation] Role of stress on gastrointestinal function and diseases. (2018)
  • Author(s): Suzuki, H.
  • Organizer: Summer School on Stress 2018, Osaka, Japan
  • Int'l Joint Research / Invited
• [Presentation] Changes of characterization in gastric cancer cells via reactive oxygen species resistance. (2018)
  • Author(s): Mori, H., Tsugawa, H., Matsuzaki, J., Masaoka, T., Kanai, T., Suzuki, H.
  • Organizer: Summer School on Stress 2018, Osaka, Japan
  • Int'l Joint Research
• [Presentation] Expression of CAPZA1, a negative regulator of CagA-degrading autophagy, is enhanced by oxidative stress-induced histone acetylation in Helicobacter pylori-infected gastric mucosa: A possible risk for gastric carcinogenesis. Esophageal, Gastric & Duodenal Disorders (EGD) Section Distinguished Abstract Plenary. (2018)
  • Author(s): Kato, C., Tsugawa, H., Saito, Y., Suzuki, H.
  • Organizer: Digestive Disease Week 2018, Washington, DC, U.S.A.
  • Int'l Joint Research
• [Presentation] Ghrelin is a useful marker of gastric mucosal atrophy after Helicobacter pylori eradication: a prospective clinical study. (2018)
  • Author(s): Mori, H., Suzuki, H., Matsuzaki, J., Masaoka, T., Igarashi, K., Kanai, T.
  • Organizer: Digestive Disease Week 2018, Washington, DC, U.S.A.
  • Int'l Joint Research
• [Presentation] Specific prediction of duodenal tumor using demographic and endoscopic characteristics: an age and sex matched case control study. (2018)
  • Author(s): Suzuki, H., Matsuzaki, J., Shimoda, M., Mori, H., Fukuhara, S., Kanai, Y., Kanai, T.
  • Organizer: Digestive Disease Week 2018, Washington, DC, U.S.A.
  • Int'l Joint Research
• [Presentation] Case study 2: Bloating & Fullness. Understanding Bloating & Fullness (2018)
  • Author(s): Suzuki, H.
  • Organizer: SEAGMA&AGIFT 4th Scientific Meeting, Singapore
  • Int'l Joint Research / Invited
• [Presentation] Case study 1: Epigastric Pain. Helicobacter pylori gastritis. (2018)
  • Author(s): Suzuki, H.
  • Organizer: SEAGMA&AGIFT 4th Scientific Meeting, Singapore
  • Int'l Joint Research / Invited
• [Presentation] Clinical evidence of Kampo medicine for functional gastrointestinal disorders (FGIDs). (2018)
  • Author(s): Suzuki, H.
  • Organizer: SEAGMA&AGIFT 4th Scientific Meeting, Singapore
  • Int'l Joint Research / Invited
• [Presentation] Role of acid suppression in FD: PPI and PCAB. (2018)
  • Author(s): Suzuki, H.
  • Organizer: APNM2018, Seoul, Korea
  • Int'l Joint Research / Invited