消化管内病原細菌特異的排除型新規分子標的療法の確立

2014 Fiscal Year Research-status Report

• Project/Area Number: 26670065
• Research Institution: Keio University
• Principal Investigator: 鈴木 秀和 慶應義塾大学, 医学部, 准教授 (70255454)
• Co-Investigator(Kenkyū-buntansha): 榊原 康文 慶應義塾大学, 理工学部, 教授 (10287427) ; 津川 仁 慶應義塾大学, 医学部, 講師 (30468483)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Keywords: FecA1 / 鉄 / H. pylori / 酸化ストレス / メトロニダゾール / NDGA

Outline of Annual Research Achievements

H. pyloriのSodB活性は、菌体内superoxide産生依存的に抗菌活性を発揮する二次除菌レジメンの1つであるメトロニダゾール(Mtz)への耐性化に寄与し、sodB遺伝子欠失により慢性感染能が消失する。SodB活性発現解析から、鉄トランスポーターFecA1依存的なSodB活性中心鉄イオン供給システムの存在を明確にし、fecA1遺伝子欠損でROS依存的宿主異物排除応答抵抗性消失による慢性感染能の破綻とMtz感受性の亢進が惹起された。さらにCOPICATプログラムを用いた解析からFecA1親和性化合物としてNordihydroguaiaretic acid (NDGA)を同定し、本化合物はH. pyloriの増殖能に影響を与えることなく、抗酸化能を有意に低下させることをin vitro inhibition zone assayで明らかとした。100uM NDGAはH. pyloriの増殖を抑制したが、50uM NDGAは菌増殖を抑制せずに、SodB活性を有意に低下させた。また、Fe2+の選択的蛍光指示薬RhoNox-1染色により、50uM NDGAは菌体内のFe2+レベルを有意に低下させた。また、50uM NDGAはH. pyloriの酸化ストレス感受性を、慢性感染能が低下することが示されているfecA1遺伝子欠損H. pyloriと同レベルまで亢進させた。Mtzは、菌体内でsuperoxideを産生し抗菌活性を示すが、SodBを過剰発現する菌株ではMtz耐性を示す。これらのMtz耐性株への50uM NDGA処理では、菌体内Fe2+レベルの低下を誘導し、SodB活性を有意に低下させた結果、Mtzへの感受性が向上した。また、これらのMtz耐性株のROS感受性もfecA1遺伝子欠損株と同レベルまで亢進させた。

Current Status of Research Progress

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

Reason

in silico探索したFecA1結合性化合物のうちでNordihydroguaiaretic acid（NDGA）が酸化ストレス感受性をシフトさせることで抗菌活性をもち、臨床応用の可能性のある化合物であることが分かった。今後は、さらに、病原性大腸菌やサルモネラ菌などのピロリ菌以外の細菌についての同様の機序が確認できるかを検討する。

Strategy for Future Research Activity

スナネズミ等をによるピロリ菌感染小動物モデルを用いて、NDGAの機能評価を実施し、in vivoでの活性を評価する。また、RhoNox-1染色を用いて、化合物スクリーニングを行うことで、より強力なFecA1阻害化合物の探索も実施する。またFecA1はグラム陰性細菌のホモログタンパク質であることから、ピロリ菌以外の大腸菌やサルモネラ菌などの他の病原細菌に対してもNDGAの効果が期待される。そこで、サルモネラ菌及び病原性大腸菌O157に対するNDGAの効果を、RhoNox-1染色並びにSOD活性測定により評価する。

Causes of Carryover

前年度に実施できなかった、ピロリ菌以外の細菌に対するNDGAの効果の検討を次年度に実施することとしたため、そのための細菌培養や分子細胞生物学的研究のための消耗品費として、次年度（平成27年度）使用額が生じた。

Expenditure Plan for Carryover Budget

FecA1阻害化合物は、ヒトのROS依存的異物排除応答依存的なピロリ菌排除を実現することが予測されるため、ピロリ菌感染スナネズミに対してFecA1阻害化合物を投与し、感染菌量を評価する。また、大腸菌やサルモネラ菌などのピロリ菌以外の消化管病原細菌に対するNDGA及びFecA1阻害化合物の効果を検討する。

Research Products

• [Journal Article] The “two-sword fencing” technique in endoscopic submucosal dissection. (2015)
  • Author(s): Nishizawa, T., Uraoka, T., Ochiai, Y., Suzuki, H., Goto, O., Fujimoto, A., Maehata, T., Kanai, T., Yahagi, N.
  • Journal Title: Clin. Endosc. Volume: 48 Pages: 85-86
  • DOI: 10.5946/ce.2015.48.1.85
  • Peer Reviewed / Open Access
• [Journal Article] Survey of clinical practice for irritable bowel syndrome in East asian countries. (2015)
  • Author(s): Fukudo, S, .Hahm, KB, Zhu, Q., Sollano, JD., Rani, AA., Syam, AF., Kachintorn, U., Suzuki, H., Kamiya, T., Joh, T., Uchiyama, K., Naito, Y., Takahashi, S., Kinoshita, Y., Ueno, F., Yamagami, H., Chan, FK., Fock, KM,, Arakawa, T.
  • Journal Title: Digestion Volume: 91 Pages: 99-109
  • DOI: 10.1159/000369078
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Editorial: Helicobacter pylori and pathogenesis. (2015)
  • Author(s): Tomoda, A., Kamiya, S., Suzuki, H.
  • Journal Title: BioMed. Res. Int. Volume: 304768 Pages: 2
  • DOI: 10.1155/2015/304768
  • Peer Reviewed / Open Access
• [Journal Article] Letter: Prompt endoscopy in Asians with uninvestigated dyspepsia - authors' reply. (2015)
  • Author(s): Chen, S.L., Gwee, K.A., Lee, J.S., Miwa, H., Suzuki, H., Guo, P., Hao, Y.T., Chen, M.H.
  • Journal Title: Aliment. Pharm. Ther. Volume: 41 Pages: 793
  • DOI: 10.1111/apt.13131
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Proton pump inhibitor alone vs proton pump inhibitor plus mucosal protective agents for endoscopic submucosal dissection-induced ulcer: a systematic review. (2015)
  • Author(s): Nishizawa, T., Suzuki, H.*, Kanai, T., Yahagi, N.
  • Journal Title: J. Clin. Biochem. Nutr. Volume: 56 Pages: 85-90
  • DOI: 10.3164/jcbn.14-101
  • Peer Reviewed / Open Access
• [Journal Article] Evidence-based clinical practice guidelines for functional dyspepsia. (2015)
  • Author(s): Miwa, H., Kusano, M., Arisawa, T., Oshima, T., Kato, M., Joh, T., Suzuki, H., Tominaga, K., Nakada, K., Nagahara, A., Futagami, S., Manabe, N., Inui, A., Haruma, K., Higuchi, K., Yakabi, K., Hongo, M., Uemura, N., Kinoshita, Y., Sugano, K., Shimosegawa, T.
  • Journal Title: J. Gastroenterol. Volume: 50 Pages: 125-139
  • DOI: 10.1007/s00535-014-1022-3
  • Peer Reviewed / Open Access
• [Journal Article] Nordihydroguaiaretic acid disrupts the antioxidant ability of Helicobacter pylori through the repression of SodB activity in vitro. (2015)
  • Author(s): Tsugawa, H., Mori, H., Matsuzaki, J., Masaoka, T., Hirayama, T., Nagasawa, H., Sakakibara, Y., Suematsu, M., Suzuki, H.
  • Journal Title: Biomed. Res. Int. Volume: 2015 Pages: 734548
  • DOI: http://dx.doi.org/10.1155/2015/734548
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Reply to letter: "Dexmedetomidine vs midazolam for gastrointestinal endoscopy". (2015)
  • Author(s): Nishizawa T, Suzuki H.
  • Journal Title: Digest. Endosc. Volume: 27 Pages: 533
  • DOI: 10.1111/den.12459.
  • Peer Reviewed / Open Access
• [Journal Article] Systematic review with meta-analysis: prompt endoscopy as the initial management strategy for uninvestigated dyspepsia in Asia. (2015)
  • Author(s): Chen, S.L., Gwee, K.A., Lee, J.S., Miwa, H., Suzuki, H., Guo, P., Hao, Y.T., Chen, M.H.
  • Journal Title: Aliment. Pharm. Ther. Volume: 41 Pages: 239-252
  • DOI: 10.1111/apt.13028.
  • Peer Reviewed / Open Access
• [Journal Article] Gastric carcinogenesis and underlying molecular mechanisms: Helicobacter pylori and novel targeted therapy. (2015)
  • Author(s): Nishizawa, T., Suzuki, H.
  • Journal Title: Biomed. Res. Int. Volume: 2015 Pages: 794378
  • DOI: http://dx.doi.org/10.1155/2015/794378
  • Peer Reviewed / Open Access
• [Journal Article] Second-look endoscopy after endoscopic submucosal dissection for gastric neoplasms: A systematic review. (2015)
  • Author(s): Nishizawa, T., Suzuki, H., Kinoshita, S., Goto, O., Kanai, T., Yahagi, N.
  • Journal Title: Digest. Endosc. Volume: 27 Pages: 279-284
  • DOI: 10.1111/den.12410.
  • Peer Reviewed / Open Access
• [Journal Article] Dexmedetomidine versus midazolam for gastrointestinal endoscopy: A meta-analysis. (2015)
  • Author(s): Nishizawa T, Suzuki, H., Sagara S, Kanai T, Yahagi N.
  • Journal Title: Digest. Endosc. Volume: 27 Pages: 8-15
  • DOI: 10.1111/den.12399.
  • Peer Reviewed / Open Access
• [Journal Article] Obscure Gastrointestinal Bleeding: Resection of a Pyogenic Granuloma of the Ileum via Double-balloon Enteroscopy. (2014)
  • Author(s): Hirata, K., Hosoe, N., Imaeda, H., Naganuma, M., Murata, H., Ueno, M., Suzuki, H., Ogata, H., Kanai, T.
  • Journal Title: Clin. J. Gastroenterol. Volume: 7 Pages: 397-401
  • Peer Reviewed / Open Access
• [Journal Article] Surveillance using trimodal imaging endoscopy after endoscopic submucosal dissection for superficial gastric neoplasia. (2014)
  • Author(s): Imaeda, H., Hosoe, N., Kashiwagi, K., Ida, Y., Nakamura, R., Suzuki, H., Saito, Y., Yahagi, N., Iwao, Y., Kitagawa, Y., Hibi, T., Ogata, H., Kanai, T.
  • Journal Title: World J Gastroenterol. Volume: 20 Pages: 16311-16317
  • DOI: 10.3748/wjg.v20.i43.16311.
  • Peer Reviewed / Open Access
• [Journal Article] Changes in the first line Helicobacter pylori eradication rates using the triple therapy – a multicenter study in the Tokyo metropolitan area (Tokyo Helicobacter pylori study group). (2014)
  • Author(s): Kawai, T., Takahashi, S., Suzuki, H., Sasaki, H., Nagahara, A., Asaoka, D., Matsuhisa, T., Masaoka, T., Nishizawa, T., Suzuki, M., Ito, M., Kurihara, N., Omata, F., Mizuno, S., Torii, A., Kawakami, K., Ohkusa, T., Tokunaga, K., Mine, T., Sakaki, N.
  • Journal Title: J. Gastroenterol. Hepatol. Volume: 29 Pages: 29-32
  • DOI: 10.4103/2153-3539.119002.
  • Peer Reviewed / Open Access
• [Journal Article] Musosal expression of aquaporin-4 in the stomach of histamine type 2 receptor knockout mice and Helicobacter pylori-infected mice. (2014)
  • Author(s): Fukuhara, S., Matsuzaki, J., Tsugawa, H., Masaoka, T., Miyoshi, S., Mori, H., Fukushima, Y., Yasui, M., Kanai, T., Suzuki, H.
  • Journal Title: J. Gastroenterol. Hepatol. Volume: 29 Pages: 53-59
  • DOI: 10.1111/jgh.12771.
  • Peer Reviewed / Open Access
• [Journal Article] Alteration of angiogenesis in Helicobacter-heilmannii-induced MALT lymphoma: interaction with c-Met and HGF. (2014)
  • Author(s): Nakamura, M., Takahashi, T., Matsui, H., Baniwa, Y., Takahashi, S., Murayama, S.Y., Serizawa, H., Suzuki, H., Hibi, T.
  • Journal Title: J. Gastroenterol. Hepatol. Volume: 29 Pages: 70-76
  • DOI: 10.1111/jgh.12776.
  • Peer Reviewed / Open Access
• [Journal Article] Clarithromycin versus metronidazole as first-line Helicobacter pylori eradication: A multicenter, prospective, randomized controlled study in Japan. (2014)
  • Author(s): Nishizawa, T., Maekawa, T., Watanabe, N., Harada, N., Hosoda, Y., Yoshinaga, M., Yoshio, T., Ohta, H., Inoue, S., Toyokawa, T., Yamashita, H., Saito, H., Kuwai, T., Katayama, S., Masuda, E., Miyabayashi, H., Kimura, T., Nishizawa, Y., Takahashi, M., Suzuki, H.
  • Journal Title: J. Clin. Gastroenterol. Volume: Epub Pages: －
  • Peer Reviewed / Open Access
• [Journal Article] Randomised clinical trial: rikkunshito in the treatment of functional dyspepsia – a multi-centre, double-blind, randomised, placebo-controlled study. (2014)
  • Author(s): Suzuki, H., Matsuzaki, J., Fukushima, Y., Suzaki, F., Kasugai, K., Nishizawa, T., Naito, Y., Hayakawa, T., Kamiya, T., Andoh, T., Yoshida, H., Tokura, Y., Nagata, H., Kobayakawa, M., Mori, M., Kato, K., Hosoda, H., Takabayashi, T., Miura, S., Uemura, N., Joh, T., Hibi, T., Tack, J.
  • Journal Title: Neurogastroenterol. Motil. Volume: 26 Pages: 950-961
  • DOI: 10.1111/nmo.12348.
  • Peer Reviewed / Open Access
• [Journal Article] Control of tip deflection by right pinkie shaft grip: right pinkie maneuver. (2014)
  • Author(s): Nishizawa, T., Uraoka, T., Suzuki, H., Ochiai, Y., Goto, O., Fujimoto, A., Maehata, T., Kanai, T., Yahagi, N.
  • Journal Title: Gastrointest. Endosc. Volume: 80 Pages: 1208
  • DOI: 10.1016/j.gie.2014.08.016.
  • Peer Reviewed / Open Access
• [Journal Article] Control of the treatment device for endoscope by left hand: two fingers method. (2014)
  • Author(s): Nishizawa, T., Uraoka, T., Suzuki, H., Kinoshita, S., Ochiai, Y., Goto, O., Fujimoto, A., Kanai, T., Yahagi, N.
  • Journal Title: Gastrointest. Endosc. Volume: 80 Pages: 1206-1207
  • DOI: 10.1016/j.gie.2014.07.063.
  • Peer Reviewed / Open Access
• [Journal Article] Greater loss of productivity among Japanese workers with gastroesophageal reflux disease (GERD) symptoms that persist vs resolve on medical therapy. (2014)
  • Author(s): Suzuki, H., Matsuzaki, J., Masaoka, T., Inadomi, J.M.
  • Journal Title: Neurogastroenterol. Motil. Volume: 26 Pages: 764-771
  • DOI: 10.1111/nmo.12319.
  • Peer Reviewed / Open Access
• [Journal Article] Ameliorative effect of mepenzolate bromide against pulmonary fibrosis. (2014)
  • Author(s): Kurotsu, S., Tanaka, K., Niino, T., Asano, T., Sugizaki, T., Azuma, A., Suzuki, H., Mizushima, T.
  • Journal Title: J. Pharmacol. Exp. Ther. Volume: 350 Pages: 79-88
  • DOI: 10.1124/jpet.114.213009.
  • Peer Reviewed / Open Access
• [Journal Article] Superiority of pulmonary administration of mepenzolate bromide over other routes as treatment for chronic obstructive pulmonary disease. (2014)
  • Author(s): Tanaka, K., Kurotsu, S., Asano, T., Yamakawa, N., Kobayashi, D., Yamashita, Y., Yamazaki, H., Ishihara, T., Watanabe, H., Maruyama, T., Suzuki, H., Mizushima, T.
  • Journal Title: Sci. Rep. Volume: 4 Pages: 4510
  • DOI: 10.1038/srep04510.
  • Peer Reviewed / Open Access
• [Journal Article] A questionnaire-based survey on the diagnosis and management of inflammatory bowel disease in East Asian countries in 2012. (2014)
  • Author(s): Hida, N., Nakamura, S., Hahm, K-B., Sollano, J.D., Zhu, Q., Rani, A.A., Syam, A.F., Kachintorn, U., Ueno, F., Joh, T., Naito, Y., Suzuki, H., Takahashi, S., Fukudo, S., Fujiwara, Y., Kinoshita, Y., Uchiyama, K., Yamaguchi, Y., Yoshida, A., Arakawa, T., Matsumoto, T., The IGICS Study Group.
  • Journal Title: Digestion Volume: 89 Pages: 88-103
  • DOI: 10.1159/000356706.
  • Peer Reviewed / Open Access
• [Journal Article] New pharmaceutical treatment of gastric MALT lymphoma: anti-angiogenesis treatment using VEGF receptor antibodies and celecoxib. (2014)
  • Author(s): Nakamura, M., Takahashi, T., Matsui, H., Takahashi, S., Murayama, SY., Suzuki, H., Tsuchimoto, K.
  • Journal Title: Curr. Pharm. Des. Volume: 20 Pages: 1097-1103
  • Peer Reviewed / Open Access
• [Journal Article] Effect of supplementation with rebamipide for Helicobacter pylori eradication therapy: a systematic review and meta-analysis. (2014)
  • Author(s): Nishizawa, T., Nishizawa, Y., Yahagi, N., Kanai, T., Takahashi, M., Suzuki, H.
  • Journal Title: J. Gastroenterol. Hepatol. Volume: 29 Pages: 20-24
  • DOI: 10.1111/jgh.12769.
  • Peer Reviewed / Open Access
• [Journal Article] Mechanisms of Helicobacter pylori antibiotic resistance and molecular testing. (2014)
  • Author(s): Nishizawa, T., Suzuki, H.
  • Journal Title: Front. Mol. Biosci. Volume: 1 Pages: 19
  • DOI: 10.3389/fmolb.2014.00019.
  • Peer Reviewed / Open Access
• [Journal Article] Propofol vs traditional sedative agents for endoscopic submucosal dissection: A systematic review. (2014)
  • Author(s): Nishizawa, T., Suzuki, H., Matsuzaki, J., Kanai, T., Yahagi, N.
  • Journal Title: Digest. Endosc. Volume: 26 Pages: 701-706
  • DOI: 10.1111/den.12342.
  • Peer Reviewed / Open Access
• [Journal Article] Does the AIMS65, a new risk score for upper gastrointestinal bleeding, work in Japan? (2014)
  • Author(s): Masaoka, T., Suzuki, H.
  • Journal Title: Digest. Endosc. Volume: 26 Pages: 331-332
  • DOI: 10.1111/den.12224.
  • Peer Reviewed / Open Access
• [Journal Article] Does bile reflux influence the progression of Barrett's esophagus to adenocarcinoma? (2014)
  • Author(s): Masaoka, T., Suzuki, H.
  • Journal Title: J. Neurogastroenterol. Motil. Volume: 20 Pages: 124-126
  • DOI: 10.5056/jnm.2014.20.1.124.
  • Peer Reviewed / Open Access
• [Journal Article] MicroRNAs in Barrett’s esophagus: Future prospects. (2014)
  • Author(s): Matsuzaki, J., Suzuki, H.
  • Journal Title: Front. Genet. Volume: 5 Pages: 69
  • DOI: 10.3389/fgene.2014.00069.
  • Peer Reviewed / Open Access
• [Presentation] LRP1-dependent host cell response in gastric epithelial cells infected with H. pylori (2015)
  • Author(s): 津川 仁、松崎 潤太郎、森 英毅、正岡 建洋、井本 正哉、末松 誠、鈴木 秀和
  • Organizer: 第88回日本細菌学会総会
  • Place of Presentation: 長良川国際会議場（岐阜県岐阜市）
  • Year and Date: 2015-03-26 – 2015-03-28
• [Presentation] Major publications in FGID and Neurogastroenterology from the West. (2015)
  • Author(s): Suzuki, H.
  • Organizer: The 4th Biennial Congress of the Asian Neurogastroenterology and Motility Association (ANMA)
  • Place of Presentation: Gurgaon, Delhi, India
  • Year and Date: 2015-02-08
  • Invited
• [Presentation] HER2陽性胃癌における酸化ストレス抵抗性がん幹細胞によるTrastuzumab耐性の獲得機構 (2014)
  • Author(s): 森英毅、津川仁、鈴木秀和
  • Organizer: JDDW 2014 （日本消化器病関連学会週間）
  • Place of Presentation: 神戸国際会議場（兵庫県神戸市）
  • Year and Date: 2014-10-23
• [Presentation] Nuclear-translocated LRP1-ICD is related to the formation of autophagolysosome associated with CagA degradation in gastric epithelial cells infected with H. pylori via induction of LAMP1 expressions. (2014)
  • Author(s): Tsugawa H., Matsuzaki J., Mori H., Masaoka T., Sato A., Imoto M., Hirayama T., Suematsu M., Suzuki H.
  • Organizer: XXVIIth International Workshop on Helicobacter & Microbiota in Inflammation & Cancer
  • Place of Presentation: Rome, Italy
  • Year and Date: 2014-09-12
• [Presentation] 胃食道逆流症(GERD)－病態・症状、診断、労働生産性から. (2014)
  • Author(s): 鈴木秀和
  • Organizer: 第56回日本平滑筋学会総会
  • Place of Presentation: 新横浜プリンスホテル（神奈川県横浜市）
  • Year and Date: 2014-08-07
  • Invited
• [Presentation] 生まれ変わる慢性胃炎の診療法～ガイドラインに基づく診療法について～ (2014)
  • Author(s): 鈴木秀和
  • Organizer: 第87回日本消化器内視鏡学会総会
  • Place of Presentation: 福岡国際会議場（福岡県福岡市）
  • Year and Date: 2014-05-16
  • Invited
• [Presentation] Efficacy and safety of rikkunshito, a Japanese herbal medicine, on the treatment of functional dyspepsia: a multi-center, double-blind, randomized, placebo-controlled study. (2014)
  • Author(s): Suzuki, H., Matsuzaki, J., Fukushima, Y., Suzaki, F., Kasugai, K., Nishizawa, T., Naito, Y., Hayakawa, T., Kamiya, T., Ando, T., Yoshida, H., Tokura, Y., Nagata, H., Kobayakawa, M., Mori, M., Kato, K., Hosoda, H., Takebayashi, T., Miura, S., Uemura, N., Joh, T., Hibi, T., Tack, J.
  • Organizer: Digestive Disease Week 2014
  • Place of Presentation: Chicago, U.S.A.
  • Year and Date: 2014-05-03
• [Presentation] 機能性ディスペプシア（FD）に対する六君子湯の有効性（多施設共同二重盲検無作為化プラセボ対照比較試験) (2014)
  • Author(s): 鈴木秀和、松崎潤太郎、福嶋康之
  • Organizer: 第100回日本消化器病学会総会
  • Place of Presentation: 東京国際フォーラム（東京都千代田区）
  • Year and Date: 2014-04-23
• [Book] eBook: OXIDATIVE STRESS AND DIETARY ANTIOXIDANTS (2014)
  • Author(s): Suzuki, H., Nishizawa, T.
  • Total Pages: 33-40
  • Publisher: Academic Press
• [Patent(Industrial Property Rights)] 宿主異物排除応答依存的除菌剤 (2014)
  • Inventor(s): 鈴木秀和、津川仁、榊原康文
  • Industrial Property Rights Holder: 鈴木秀和、津川仁、榊原康文
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2014-106192
  • Filing Date: 2014-05-22