Researcher Profile and Settings

researchmap

https://researchmap.jp/read0139879

Research Areas

• Life sciences / Clinical pharmacy

Research Experience

• 同志社大学工学部環境システム学科特別研究員, 2007 - 2008
• 国立国際医療センター研究所臨床薬理研究部流動研究員, 2003 - 2007

Education

• Hoshi University, 薬学研究科, 薬学専攻, 2003/04 - 2006/03
• Hoshi University, 薬学研究科, 薬学専攻, 2001/04 - 2003/03

Awards

• 腸管出血性大腸菌感染症研究会 奨励賞
  2012

Published Papers

• CaMKII-dependent non-canonical RIG-I pathway promotes influenza virus propagation in the acute-phase of infection
  Shinichiro Hama; Miho Watanabe-Takahashi; Hiroki Nishimura; Jumpei Omi; Masakazu Tamada; Takashi Saitoh; Katsumi Maenaka; Yuta Okuda; Aoi Ikegami; Asami Kitagawa; Koudai Furuta; Kana Izumi; Eiko Shimizu; Takashi Nishizono; Makoto Fujiwara; Tomohiro Miyasaka; Shigeo Takamori; Hiroshi Takayanagi; Keizo Nishikawa; Toshihiko Kobayashi; Noriko Toyama-Sorimachi; Makoto Yamashita; Toshiya Senda; Takatsugu Hirokawa; Haruhiko Bito; Kiyotaka Nishikawa
  mBio, American Society for Microbiology, 27 Nov. 2024, Scientific journal
  

  URLDOI URL
• A tetravalent peptide efficiently inhibits the intestinal toxicity of heat-labile enterotoxin by targeting the receptor-binding region of the B-subunit pentamer
  Miho Watanabe-Takahashi; Tetsuya Tanigawa; Takashi Hamabata; Kiyotaka Nishikawa
  Biochemical and Biophysical Research Communications, Elsevier BV, 734 150769 - 150769, Nov. 2024, Scientific journal
  

  DOI URL
• Tailored multivalent peptide targeting the B-subunit pentamer of Cholera toxin inhibits its intestinal toxicity by inducing aberrant transport of the toxin in cells
  Watanabe-Takahashi M; Kumoi K; Yamamoto H; Shimizu E; Motoyama J; Hamabata T; Nishikawa K
  Biochem. Biophys. Res. Commun., 716 149991 , Apr. 2024, Scientific journal
  

  DOI URL
• A tailored tetravalent peptide displays dual functions to inhibit amyloid β production and aggregation.
  Sato W; Watanabe-Takahashi M; Murata T; Utsunomiya-Tate N; Motoyama J; Anzai M; Ishihara S; Nishioka N; Uchiyama H; Togashi J; Nishihara S; Kawasaki K; Saito T; Saido T. C; Funamoto S*; Nishikawa K*
  Commun. Biol., 6(1)383(1) 383 - 383, Apr. 2023, Scientific journal
  

  DOI URL
• Novel multivalent S100A8 inhibitory peptides attenuate tumor progression and metastasis by inhibiting the TLR4-dependent pathway.
  Atsuko Deguchi; Miho Watanabe-Takahashi; Taishi Mishima; Tsutomu Omori; Umeharu Ohto; Nobuto Arashiki; Fumio Nakamura; Kiyotaka Nishikawa; Yoshiro Maru
  Cancer gene therapy, 30 973 - 984, 17 Mar. 2023, Scientific journal
  

  DOI URL
• Hepatitis B Virus Utilizes a Retrograde Trafficking Route via the Trans-Golgi Network to Avoid Lysosomal Degradation.
  Ying-Yi Li; Kazuyuki Kuroki; Tetsuro Shimakami; Kazuhisa Murai; Kazunori Kawaguchi; Takayoshi Shirasaki; Kouki Nio; Saiho Sugimoto; Tomoki Nishikawa; Hikari Okada; Noriaki Orita; Hideo Takayama; Ying Wang; Phuong Doan Thi Bich; Astuya Ishida; Sadahiro Iwabuchi; Shinichi Hashimoto; Takeshi Shimaoka; Noriko Tabata; Miho Watanabe-Takahashi; Kiyotaka Nishikawa; Hiroshi Yanagawa; Motoharu Seiki; Kouji Matsushima; Taro Yamashita; Shuichi Kaneko; Masao Honda
  Cellular and molecular gastroenterology and hepatology, 15(3) 533 - 558, 2023, Scientific journal
  

  DOI URL
• A tetravalent peptide that binds to the RANK-binding region of TRAF6 via a multivalent interaction efficiently inhibits osteoclast differentiation.
  Anzai M; Watanabe-Takahashi M; Kawabata H; Mizuno S; Taguchi Y; Inoue J; Nishikawa K
  Biochem. Biophys. Res. Commun., 636(Pt 1) 178 - 183, Nov. 2022, Scientific journal
  

  DOI URL
• Development of a novel tetravalent peptide that absorbs subtilase cytotoxin by targeting the receptor-binding B-subunit
  Shinichiro Hama; Miki Nakahara; Miho Watanabe-Takahashi; Eiko Shimizu; Hiroyasu Tsutsuki; Kinnosuke Yahiro; Kiyotaka Nishikawa
  Biochemical and Biophysical Research Communications, Elsevier BV, 629(12) 95 - 100, Sep. 2022, Scientific journal
  

  DOI URL
• A unique peptide-based pharmacophore identifies an inhibitory compound against the A-subunit of Shiga toxin
  Watanabe-Takahashi M; Senda M; Yoshino R; Hibino M; Hama S; Terada T; Shimizu K; Senda T; Nishikawa K
  Scientific Reports, Springer Science and Business Media LLC, 12:11443(1), Jul. 2022, Scientific journal
  

  URLDOI URL
• Identification of a peptide motif that potently inhibits two functionally distinct subunits of Shiga toxin
  Miho Watanabe-Takahashi; Masakazu Tamada; Miki Senda; Masahiro Hibino; Eiko Shimizu; Akiko Okuta; Atsuo Miyazawa; Toshiya Senda; Kiyotaka Nishikawa
  Communications Biology, Springer Science and Business Media LLC, 4(1), Dec. 2021, Scientific journal
  

  DOI URLScopus URLScopus Citedby URL
• A nontoxigenic form of Shiga toxin 2 suppresses the production of amyloid β by altering the intracellular transport of amyloid precursor protein through its receptor-binding B-subunit
  Waka Sato; Miho Watanabe-Takahashi; Takashi Hamabata; Koichi Furukawa; Satoru Funamoto; Kiyotaka Nishikawa
  Biochemical and Biophysical Research Communications, Elsevier BV, 557 247 - 253, Jun. 2021, Scientific journal
  

  DOI URL
• The Inducible Amphisome Isolates Viral Hemagglutinin and Defends Against Influenza A Virus Infection.
  Omi J; Watanabe-Takahashi M; Igai K; Shimizu E; Ching-Yi Tseng; Miyasaka, T; Waku T; Hama S; Nakanishi R; Goto Y; Nishino Y; Miyazawa A; Natori Y; Yamashita M; Kiyotaka Nishikawa K
  Nat. Commun., 11(1)162(1) doi; 10.1038/s41467-019-13974- - 162, Jan. 2020, Scientific journal
  

  DOI URL
• Synthetic construction of sugar-amino acid hybrid polymers involving globotriaose or lactose and evaluation of their biological activities against Shiga toxins produced by Escherichia coli O157:H7
  Koji Matsuoka; Kiyotaka Nishikawa; Yusuke Goshu; Tetsuo Koyama; Ken Hatano; Takahiko Matsushita; Miho Watanabe-Takahashi; Yasuhiro Natori; Daiyo Terunuma
  Bioorganic & Medicinal Chemistry, Elsevier BV, 26(22) 5792 - 5803, Dec. 2018, Scientific journal
  

  DOI URL
• Exosome-associated Shiga toxin 2 is released from cells and causes severe toxicity in mice.
  Miho Watanabe-Takahashi; Shinji Yamasaki; Masayuki Murata; Fumi Kano; Jun Motoyama; Jyoji Yamate; Jumpei Omi; Waka Sato; Hirofumi Ukai; Kentaro Shimasaki; Masaya Ikegawa; Miwa Tamura-Nakano; Ryohei Yanoshita; Yuri Nishino; Atsuo Miyazawa; Yasuhiro Natori; Noriko Toyama-Sorimachi; Kiyotaka Nishikawa
  Scientific reports, 8(1) 10776 - 10776, 17 Jul. 2018, Scientific journal
  

  DOI URL
• The pleckstrin homology domain of p210 BCR-ABL interacts with cardiolipin to regulate its mitochondrial translocation and subsequent mitophagy.
  Shimasaki K; Watanabe-Takahashi M; Umeda M; Funamoto S; Saito Y; Noguchi N; Kumagai K; hanada K; Tsukahara F; Maru Y; Shibata N; Naito M; Nishikawa K
  Genes to Cells, 23(1) 22 - 34, Jan. 2018
• Acquired Resistance to Shiga toxin-induced apoptosis by loss of CD77 expression in Human myelogenous leukemia cell line, THP-1.
  Takahashi M; Hattori T; Watanabe-Takahashi M; Nishikawa K; Nito M
  Biological and Pharmaceutical Bulletin, 41;9 1475 - 1479, 2018
• Affinity-Based Screening of Tetravalent Peptides Identifies SubtypeS-elective Neutralizers of Shiga Toxin 2d, a Highly Virulent Subtype, by Targeting a Unique Amino Acid Involved in Its Receptor Recognition
  Takaaki Mitsui; Miho Watanabe-Takahashi; Eiko Shimizu; Baihao Zhang; Satoru Funamoto; Shinji Yamasaki; Kiyotaka Nishikawa
  INFECTION AND IMMUNITY, AMER SOC MICROBIOLOGY, 84(9) 2653 - 2661, Sep. 2016, Scientific journal
  

  DOI URL
• M-COPA, a novel Golgi system disruptor, suppresses apoptosis induced by Shiga toxin
  Takayuki Hattori; Miho Watanabe-Takahashi; Isamu Shiina; Yoshimi Ohashi; Shingo Dan; Kiyotaka Nishikawa; Takao Yamori; Mikihiko Naito
  GENES TO CELLS, WILEY, 21(8) 901 - 906, Aug. 2016, Scientific journal
  

  DOI URL
• Development of a Novel Tetravalent Synthetic Peptide That Binds to Phosphatidic Acid
  Rina Ogawa; Kohjiro Nagao; Kentaro Taniuchi; Masaki Tsuchiya; Utako Kato; Yuji Hara; Takehiko Inaba; Toshihide Kobayashi; Yoshihiro Sasaki; Kazunari Akiyoshi; Miho Watanabe-Takahashi; Kiyotaka Nishikawa; Masato Umeda
  PLOS ONE, PUBLIC LIBRARY SCIENCE, 10(7), Jul. 2015, Scientific journal
  

  DOI URL
• Identification of a Wide Range of Motifs Inhibitory to Shiga Toxin by Affinity-Driven Screening of Customized Divalent Peptides Synthesized on a Membrane
  Mihoko Kato; Miho Watanabe-Takahashi; Eiko Shimizu; Kiyotaka Nishikawa
  APPLIED AND ENVIRONMENTAL MICROBIOLOGY, AMER SOC MICROBIOLOGY, 81(3) 1092 - 1100, Feb. 2015, Scientific journal
  

  DOI URL
• Proteasome inhibitors prevent cell death and prolong survival of mice challenged by Shiga toxin
  Takayuki Hattori; Miho Watanabe-Takahashi; Nobumichi Ohoka; Takashi Hamabata; Koichi Furukawa; Kiyotaka Nishikawa; Mikihiko Naito
  FEBS OPEN BIO, ELSEVIER SCIENCE LONDON, 5 605 - 614, 2015, Scientific journal
  

  DOI URL
• Identification of a Peptide-Based Neutralizer That Potently Inhibits Both Shiga Toxins 1 and 2 by Targeting Specific Receptor-Binding Regions
  Kazue Tsutsuki; Miho Watanabe-Takahashi; Yasuaki Takenaka; Eiji Kita; Kiyotaka Nishikawa
  INFECTION AND IMMUNITY, AMER SOC MICROBIOLOGY, 81(6) 2133 - 2138, Jun. 2013, Scientific journal
  

  DOI URL
• Substrate ectodomain is critical for substrate preference and inhibition of γ-secretase
  Satoru Funamoto; Toru Sasaki; Seiko Ishihara; Mika Nobuhara; Masaki Nakano; Miho Watanabe-Takahashi; Takashi Saito; Nobuto Kakuda; Tomohiro Miyasaka; Kiyotaka Nishikawa; Takaomi C. Saido; Yasuo Ihara
  Nature Communications, 4, 2013, Scientific journal
  

  DOI URL
• An Orally Applicable Shiga Toxin Neutralizer Functions in the Intestine To Inhibit the Intracellular Transport of the Toxin
  Miho Watanabe-Takahashi; Toshio Sato; Taeko Dohi; Noriko Noguchi; Fumi Kano; Masayuki Murata; Takashi Hamabata; Yasuhiro Natori; Kiyotaka Nishikawa
  INFECTION AND IMMUNITY, AMER SOC MICROBIOLOGY, 78(1) 177 - 183, Jan. 2010, Scientific journal
  

  DOI URL
• Yip1A regulates the COPI-independent retrograde transport from the Golgi complex to the ER
  Fumi Kano; Shinobu Yamauchi; Yumi Yoshida; Miho Watanabe-Takahashi; Kiyotaka Nishikawa; Nobuhiro Nakamura; Masayuki Murata
  JOURNAL OF CELL SCIENCE, COMPANY OF BIOLOGISTS LTD, 122(13) 2218 - 2227, Jul. 2009, Scientific journal
  

  DOI URL
• Regulation of hepatic cholesterol synthesis by a novel protein (SPF) that accelerates cholesterol biosynthesis
  Norihito Shibata; Kou-ichi Jishage; Makoto Arita; Miho Watanabe; Yosuke Kawase; Kiyotaka Nishikawa; Yasuhiro Natori; Hiroyasu Inoue; Hitoshi Shimano; Nobuhiro Yamada; Masafumi Tsujimoto; Hiroyuki Arai
  FASEB JOURNAL, FEDERATION AMER SOC EXP BIOL, 20(14) 2642 - +, Dec. 2006, Scientific journal
  

  DOI URL
• A multivalent peptide library approach identifies a novel Shiga toxin inhibitor that induces aberrant cellular transport of the toxin
  Kiyotaka Nishikawa; Miho Watanabe; Eiji Kita; Katsura Igai; Kazumi Omata; Michael B. Yaffe; Yasuhiro Natori
  FASEB JOURNAL, FEDERATION AMER SOC EXP BIOL, 20(14) 2597 - +, Dec. 2006, Scientific journal
  

  DOI URL
• Development of dialyzer with immobilized glycoconjugate polymers for removal of Shiga-toxin
  A Miyagawa; M Watanabe; K Igai; MCZ Kasuya; Y Natori; K Nishikawa; K Hatanaka
  BIOMATERIALS, ELSEVIER SCI LTD, 27(17) 3304 - 3311, Jun. 2006, Scientific journal
  

  DOI URL
• Structural analysis of the interaction between Shiga toxin B subunits and linear polymers bearing clustered globotriose residues
  M Watanabe; K Igai; K Matsuoka; A Miyagawa; T Watanabe; R Yanoshita; Y Samejima; D Terunuma; Y Natori; K Nishikawa
  INFECTION AND IMMUNITY, AMER SOC MICROBIOLOGY, 74(3) 1984 - 1988, Mar. 2006, Scientific journal
  

  DOI URL
• Identification of the optimal structure required for a Shiga toxin neutralizer with oriented carbohydrates to function in the circulation
  K Nishikawa; K Matsuoka; M Watanabe; K Igai; K Hino; K Hatano; A Yamada; N Abe; D Terunuma; H Kuzuhara; Y Natori
  JOURNAL OF INFECTIOUS DISEASES, UNIV CHICAGO PRESS, 191(12) 2097 - 2105, Jun. 2005, Scientific journal
• Oral therapeutic agents with highly clustered globotriose for treatment of Shiga toxigenic Escherichia coli infections
  M Watanabe; K Matsuoka; E Kita; K Igai; N Higashi; A Miyagawa; T Watanabe; R Yanoshita; Y Samejima; D Terunuma; Y Natori; K Nishikawa
  JOURNAL OF INFECTIOUS DISEASES, UNIV CHICAGO PRESS, 189(3) 360 - 368, Feb. 2004, Scientific journal

MISC

• 抗A型インフルエンザウイルス活性を示す誘導性アンフィソームの形成機構の解明
  村上 絵理; 福田 協平; 井深 健太郎; 高橋 美帆; 柴田 剛明; 近江 純平; 可野 邦行; 河野 望; 青木 淳賢; 西川 喜代孝
  日本生化学会大会プログラム・講演要旨集, (公社)日本生化学会, 94回 [P - 903], Nov. 2021
• 4価型人工ペプチドを用いたホスファチジン酸結合プローブの開発
  小川 莉奈; 長尾 耕治郎; 原 雄二; 田村 朋則; 浜地 格; 井上 飛鳥; 青木 淳賢; 高橋 美帆; 西川 喜代孝; 梅田 真郷
  脂質生化学研究, 日本脂質生化学会, 57 170 - 173, May 2015
• ホスファチジン酸結合性を有する4価型人工ペプチドの開発
  小川莉奈; 宮崎俊秀; 長尾耕治郎; 原雄二; 佐々木善浩; 秋吉一成; 稲葉岳彦; 小林俊秀; 高橋美帆; 西川喜代孝; 梅田真郷
  日本生化学会大会(Web), (公社)日本生化学会, 87th 2T09A-10(2P-071) (WEB ONLY) - 10], Oct. 2014
  

  URL
• Synthesis and biological evaluation of glycopolymer as Shiga toxin neutralizer
  K Matsuoka; A Miyagawa; K Nishikawa; M Watanabe; Y Natori; E Kita; T Koyama; K Hatano; D Terunuma
  GLYCOBIOLOGY, OXFORD UNIV PRESS INC, 14(11) 1194 - 1194, Nov. 2004, Summary international conference

Presentations

• Stx2fのAならびにBサブユニットを 標的とした阻害薬開発
  髙野 祐輝; 高橋 美帆; 六車 健太; 中筋 圭穂; 石嶋 希; 伊豫田 淳; 西川 喜代孝
  第26回腸管出血性大腸菌感染症研究会, Nov. 2024
• 個体強毒性発現に関わるStx2aの 細胞外放出機構におけるGSK3βの関与
  寺島 俊結; 髙橋 美帆; 平田 慶治; 尾上 舞衣; 西川 喜代孝
  第26回腸管出血性大腸菌感染症研究会, Nov. 2024
• Shiga toxin 2 (Stx2)の細胞内輸送制御による個体毒性の抑制
  髙橋 美帆; 平田 慶治; 尾上 舞衣; 西川 喜代孝
  第25回腸管出血性大腸菌感染症研究会, 12 Sep. 2023, Oral presentation
• 受容体結合部位を標的とした新規ペプチド性Subtilase cytotoxin阻害薬の開発
  中原美樹; 高橋美帆; 及川英子; 八尋錦之助; 西川喜代孝
  第２２回腸管出血性大腸菌感染症研究会, 2018
• 酵素活性部位を標的とした新規ペプチド性Stx阻害薬の開発
  日比野雅大; 高橋美帆; 玉田真一; 千田美紀; 奥田明子; 宮澤淳夫; 千田俊哉; 西川喜代孝
  第２２回腸管出血性大腸菌感染症研究会, 2018
• Exosome-associated Shiga toxin 2 is released from cells and causes severe toxicity in mice.
  Miho Watanabe-Takahashi; Shinji Yamasaki; Masayuki Murata; Fumi Kano; Jun Motoyama; Jyoji Yamate; Jumpei Omi; Waka Sato; Hirofumi Ukai; Kentaro Shimasaki; Masaya Ikegawa; Miwa Tamura-Nakano; Ryohei Yanoshita; Yuri Nishino; Atsuo Miyazawa; Yasuhiro Natori; Noriko Toyama-Sorimachi; Kiyotaka Nishikawa
  VTEC2018, 2018
• Structural analysis of the interaction between Shiga toxin B subunits and linear polymers bearing clustered globotriose residues
  VTEC2006, 2006, Poster presentation
• Carbohydorate-containing linear polymers that neutralize Shiga toxin
  VTEC 2003, 2003, Poster presentation

Industrial Property Rights

• Patent right
  P210 PH結合ペプチドおよび慢性骨髄性白血病治療薬
  西川喜代孝, 高橋美帆, 島崎健太朗, 長田雅也
  特願2018-210822
• Patent right
  Stx1毒性阻害ペプチドおよびStx1に起因する疾患の治療薬
  西川喜代孝, 高橋美帆, 津々木一恵
  特願2010-019315, 特願2010-019315
• Patent right
  Stx2阻害４価ペプチドおよびこのStx2阻害４価ペプチドを含む治療薬
  西川喜代孝, 高橋美帆, 三井貴瑛, 山崎伸二
  特願2013-13746, 特願2013-13746
• Patent right
  LT阻害４価ペプチドおよびETEC感染症治療薬
  西川喜代孝, 高橋美帆, 谷川哲也
  特願2014-128632, 特願2014-128632
• Patent right
  CT阻害４価ペプチドおよびコレラ治療薬
  西川喜代孝, 高橋美帆, 山本洋, 濱端崇
  特願2013-51032, 特願2014-050828
• Patent right
  ＣａＭＫII阻害ペプチドおよびこれを含有するＣａＭＫII阻害剤
  西川喜代孝, 高橋美帆, 西村浩輝, 高柳広, 尾藤晴彦
  特許第5754008号, 特許第5754008号
• Patent right
  ヘマグルチニン結合ペプチド、および、これを含むインフルエンザウイルス感染症の予防・治療薬
  西川喜代孝, 高橋美帆, 近江純平
  特願2016-164971, 特願2016-164971
• Patent right
  Stx毒性阻害ペプチドおよびStxに起因する疾患の治療薬
  西川喜代孝, 高橋美帆, 津々木一恵
  特許第5635779号, 特許第5635779号
• Patent right
  破骨細胞分化制御ペプチド、および、破骨細胞分化に関連する疾患の含有する治療薬
  西川喜代孝, 高橋美帆, 安西聖敬, 川端宏, 水野沙緒利
  特願2018-163521, 特願2018-163521
• Patent right
  抗インフルエンザウイルス活性ペプチドおよびインフルエンザウイルス感染症の予防・治療薬
  西川喜代孝, 高橋美帆, 近江純平, 濱信一郎
  特願2018-163326, 特願2018-163326

Research Projects

• Molecular mechanism underlying the production of exosome-associated Shiga toxin 2a, which causes severe toxicity in mice
  高橋 美帆
  O157:H7等の腸管出血性大腸菌の主要な病原因子であるShiga toxin（Stx)にはStx1とStx2のサブタイプが存在する。両者の細胞毒性は同程度だが、個体レベルではStx2の方が数百倍強毒であることが知られている。Stx2は臨床的にも症状の重篤化と深く関係することから、Stx2に対する阻害剤開発は急務である。これまでに、Stx2に特徴的な細胞内小胞輸送機構が存在し、本経路を経由してStx2が積極的に細胞外に放出されること、このとき一部のStx2はエクソソーム結合型Stx2（exo-Stx2)であること、さらにこのexo-Stx2がマウス個体での強毒性に直接関与していることを明らかにした。また、複数種のプロテインキナーゼがStx2の細胞外遊離およびexo-Stx2産生に関わる細胞内輸送に寄与している可能性が考えられた。そこで本研究ではGSK3βに着目し、１）Stx感受性Vero細胞にsiRNAを添加しGSK3βをノックダウンした際の、Stx2の細胞外放出量をI125標識Stx2を用いて測定した。その結果、GSK3βをノックダウンした場合に有意にStx2の細胞外放出量が減弱することが示された。２）GSK3β阻害剤Indirubinを用いて１）と同様の検討をしたところ、有意にStx2の細胞外放出量が抑制された。そこで次に、３）マウスを用いてStx2致死性に対するindirubinの効果を検討した。マウス尾静脈からStx2を 3-10 ng投与すると4日以内に死亡した。そこでIndirubin (10mg/kg)を腹腔内投与したのち、Stx2を尾静脈投与した。その結果、Indirubin投与群では明らかにStx2の個体毒性が抑制されマウスは延命していることが示された。以上よりGSK3betaはマウス個体でみられるexo-Stx2による強毒性発現に寄与すると考えられる。, Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, 2023/04 -2026/03, Grant-in-Aid for Scientific Research (C), Doshisha University
• Development of a new therapeutic agent that inhibits the production of exosome-associated Shiga toxin 2
  Takahashi Miho
  After being incorporated into target cells, Shiga toxin 2 (Stx2) was actively released from the cells, and part of released Stx2a was associated with exosomes (refer to as exo-Stx2). To determine the intracellular molecules that regulate the production of exo-Stx2, we examined the effect of knockdown of Rabs (Rab11a, Rab11b, Rab27a, Rab27b, and Rab35) on the release of Stx2 in the culture medium. The results showed that the release of Stx2 was inhibited by Rab11b knockdown but not by the other Rabs knockdown, indicating that exo-Stx2 is released by Rab11b dependent machinery. By using a kinase substrate peptide array, we analyzed the hundreds of protein kinases activations in Stx treated cells, and found that the types of activated protein kinases were different between the case with Stx1 and Stx2 in the intracellular transport of Stx., Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, 2018/04 -2021/03, Grant-in-Aid for Scientific Research (C), Doshisha University
• Development of a novel Shiga toxin inhibitor that targeting of intracellular transport of the toxin
  Takahashi Miho
  We synthesized substrate peptides of various kinases on a slide glass and using this, we analyzed the activities of kinases stimulated by Stx1a in the presence or absence of Stx inhibitors (MMA-tet, FRA-tet).We established a new multivalent peptide library screening by using tetravalent peptides synthesized on a cellulose membrane., Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, 2015/04 -2018/03, Grant-in-Aid for Scientific Research (C), Doshisha University
• Development peptide-based compounds which bind to the specific receptor-binding region of Shiga toxin B subunit
  TAKAHASHI Miho
  To identify the peptide probes that specifically bind to one of the receptor-binding site 2 of Shiga toxin1 (Stx1), we established a novel screening system using the multivalent SPOT peptide array. We synthesized total 361 SPOT peptides on the cellulose membrane, and screened the peptides based on the binding affinities against the Stx1B subunit or Stx1 site 2 mutant (G62A). Finally, we identified 11 peptides that specifically bound to the Stx1B subunit, but not to G62A mutant. Furthermore, we found the 4 peptides among them (PQA-tet, KGA-tet, VIA-tet, YTA-tet) effectively inhibited the cytotoxicity for Stx1 in Vero cells. Thus, the multivalent SPOT peptide array technique may provide a powerful tool to identify a series of effective peptide-based Stx neutralizers which target a specific receptor binding region., Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, 2010 -2012, Grant-in-Aid for Young Scientists (B), Doshisha University
• An orally applicable Shiga toxin neutralizer functions in the intestine to inhibit the intracellular transport of the toxin.
  TAKAHASHI Miho
  We found that PPP-tet, a peptide-based Stx2 inhibitor, completely inhibited fluid accumulation in the rabbit ileum caused by the direct injection of Stx2. We also found that PPP-tet accumulated in the ileal epithelial cells only through its formation of a complex with Stx2. The complex in cultured epithelial cells blocked the intracellular transport of Stx2 from the Golgi apparatus to the endoplasmic reticulum, the process that is essential for Stx2 cytotoxicity. Our results reveal that PPP-tet functions as a potent Stx neutralizer in the intestine by altering the intracellular transport of Stx2 in epithelial cells., Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, 2008 -2009, Grant-in-Aid for Young Scientists (B), Doshisha University

Research Seeds

• Development of a novel therapeutic agent for CML that targets a PH domain of p210-BCR-ABL

  Research Fields: Medical and Pharmaceutical Sciences

  Summary: The tyrosine kinase inhibitors (TKIs) for CML treatment causes drug-resistance, and shows only a poor effect against CML stem cells. In CML cells and CML stem cells, it has been shown that a high level of ROS production is sustained to promote the proliferation of these cells. Thus, the compound targeting the PH domain to regulate its function can be a novel type of therapeutic agent against CML.
• A new technology identifying a specific peptide-based inhibitor for a protein kinase.

  Research Fields: Medical and Pharmaceutical Sciences, Life Sciences

  Affiliation/Name: Kiyotaka Nishikawa:Miho Takahashi

  

  Patent Information: 特願2014-128632, 特願2010-019731, 特願2010-019728