Challenge to the accurate detection of minimal drug resistant cells

• Project/Area Number: 25670184
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Experimental pathology
• Research Institution: Hokkaido University
• Principal Investigator: OHBA Yusuke 北海道大学, 医学(系)研究科(研究院), 教授 (30333503)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: バイオイメージング / 腫瘍 / 薬剤耐性 / フローサイトメーター / FRET / CML / 蛍光寿命

Outline of Final Research Achievements

In Philadelphia chromosome-positive leukemia, growth and survival of leukemia cells solely depend on constitutive tyrosine kinase activity of new fusion gene product BCR-ABL formed by chromosome reciprocal translocation between chromosomes 9 and 22. Therefore, the treatment for this disease has been radically improved by the introduction of the specific tyrosine kinase inhibitor of BCR-ABL. However, it is desired to develop new technologies that enable the detection of a minimal number of drug-resistant cells because substantial patients experience drug resistance regardless of the presence or absence of past and current treatment. We have previously developed a novel technique to detect relatively small number of drug resistant cells. Here, we challenged to centuplicate the detection ability by the improvement of the FRET biosensor, the detection method and the analytical method. As a result, we obtained 162-fold improvement throughout this study.

Research Products

• [Journal Article] Fluorescent protein-based biosensors to visualize signal transduction beneath the plasma membrane (2015)
  • Author(s): Y. Fujioka, et al.
  • Journal Title: Anal. Sci. Volume: in press
  • Peer Reviewed
• [Journal Article] Molecular role of RNF43 in canonical and noncanonical Wnt signaling (2015)
  • Author(s): T. Tsukiyama,et al.
  • Journal Title: Mol. Cell. Biol. Volume: in press Issue: 11 Pages: 2007-2023
  • DOI: 10.1128/mcb.00159-15
  • Peer Reviewed / Open Access
• [Journal Article] CRK adaptor protein induces epithelial-mesenchymal transition and metastasis of bladder cancer cells via HGF/c-Met feedback loop. (2015)
  • Author(s): Matsumoto R, Tsuda M, Wang L, Maishi N, Abe t, Kimura T, Tanino M, Nishihara H, Hida K, Ohba Y, Shinohara N, Nonomura K, Tanaka S.
  • Journal Title: Cancer Sci Volume: in press Issue: 6 Pages: 709-17
  • DOI: 10.1111/cas.12662
  • Peer Reviewed / Open Access
• [Journal Article] p18/Stathmin1 is regulated by miR-31 in ovarian cancer in response to taxane (2015)
  • Author(s): M.K. Hassan,et al.
  • Journal Title: Oncoscience Volume: in press
  • Peer Reviewed
• [Journal Article] Inhibition of multidrug transporter in tumor endothelial cells enhances antiangiogenic effects of low-dose metronomic paclitaxel (2015)
  • Author(s): 1.Akiyama K., Maishi N., Ohga N., Hida Y., Ohba Y., Alam Mohammad Towfik, Kawamoto T., Ohmura H., Yamada K., Torii C., Shindoh M. and *Hida K.
  • Journal Title: Am J Pathol Volume: 185(2) Issue: 2 Pages: 572-580
  • DOI: 10.1016/j.ajpath.2014.10.017
  • Peer Reviewed / Open Access
• [Journal Article] Apoptosis and Molecular Targeting Therapy in Cancer (2014)
  • Author(s): M. Hassan, et al.
  • Journal Title: Biomed Res. Int. Volume: 2014 Pages: 150845-150845
  • DOI: 10.1155/2014/150845
  • Peer Reviewed
• [Journal Article] Agonist-promoted ubiquitination differentially regulates receptor trafficking of endothelin type A and type B receptors. (2014)
  • Author(s): Terada K., Horinouchi T., Fujioka Y., Higashi T., Nepal P., Horiguchi M., Sarita K., Hatate C., Hoshi A., Harada T., Mai Y., Ohba Y. and Miwa S.
  • Journal Title: J. Biol. Chem. Volume: 289 Issue: 51 Pages: 35283-35295
  • DOI: 10.1074/jbc.m113.544171
  • NAID: 120005549588
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Lysosomal interaction of Akt with Phafin2 : a critical step in the induction of autophagy (2014)
  • Author(s): M. Matsuda-Lennikov, F. Suizu, N. Hirata, M. Hashimoto, K. Kimura, T. Nagamine, Y. Fujioka, Y. Ohba, T. Iwanaga, and M. Noguchi
  • Journal Title: PLoS One Volume: 9 Issue: 5 Pages: 671-685
  • DOI: 10.1093/neuonc/not239
  • NAID: 120005385325
  • Peer Reviewed
• [Journal Article] Histone deacetylase inhibitors sensitize lung cancer cells to hyperthermia : involvement of Ku70/SirT-1 in thermo-protection (2014)
  • Author(s): M.K. Hassan, H. Watari, A.E. Salah-EldinAE, A.S. Sultan, Z. Mohamed, Y. Fujioka, Y. Ohba, N. Sakuragi
  • Journal Title: PLoS One Volume: 9 Issue: 4 Pages: e94213-e94213
  • DOI: 10.1371/journal.pone.0094213
  • NAID: 120005462418
  • Peer Reviewed / Open Access
• [Journal Article] ［特集］細胞の少数制と多様性に挑むシングルセルアナリシス―患者血液とFRETバイオセンサーを用いた薬効効果予測 (2014)
  • Author(s): 大場雄介
  • Journal Title: 生体の科学 Volume: 65 Pages: 171-175
• [Journal Article] A Ca2+-dependent signaling circuit regulates influenza A virus internalization and infection (2013)
  • Author(s): Fujioka Y, Tsuda M, Nanbo A, Hattori T, Sasaki J, Sasaki T, Miyazaki T, Ohba Y
  • Journal Title: Nature Co㎜unnications Volume: 4 Issue: 1 Pages: 2-763
  • DOI: 10.1038/ncomms3763
  • Peer Reviewed
• [Journal Article] Beneficial innate signaling interference for anti-bacterial responses by a TLR-mediated enhancement of the MKP-IRF3 axis (2013)
  • Author(s): Negishi H, Matsuki K, Endo N, Sarashina H. Miki S, Matsuda A, Fukazawa K, Taguchi-Atarashi N, Ikushima H, Yanai H, Nishio J, Honda K, Fujioka Y, Ohba Y, Noda T, Taniguchi S, Nishida E, Zhang Y, Chi H, Flavell RA, Taniguchi T
  • Journal Title: Proc Nat1 Acad Sci U S A Volume: 49 Issue: 49 Pages: 19884-19889
  • DOI: 10.1073/pnas.1320145110
  • Peer Reviewed
• [Journal Article] Inhibition of influenza A virus infection by Galectin-9 (2013)
  • Author(s): T. Hattori, J. Maruyama, Y. Fujioka, Y. Nakayama, Y. Ohba, T. Niki, T. Arikawa, T. Miyazaki, M. Hirashima, and H. Kida a
  • Journal Title: Jpn. J. Vet. Res. Volume: 61 Pages: 5-18
  • NAID: 120005289519
  • Peer Reviewed
• [Journal Article] All members of the EPI64 subfamily of TBC/RabGAPs also have GAP activities toward Ras (2013)
  • Author(s): Hiroyuki Nagai
  • Journal Title: J. Biochem Volume: 153 Issue: 3 Pages: 283-288
  • DOI: 10.1093/jb/mvs147
  • Peer Reviewed
• [Presentation] FRET イメージングによる BMAL1-CLOCK 相互作用の解析 (2015)
  • Author(s): 西出真也他
  • Organizer: 第92回日本生理学会
  • Place of Presentation: 神戸国際会議場（兵庫県神戸市）
  • Year and Date: 2015-03-23
• [Presentation] Development of FRET-based biosensors for detection of CML cells resistant to molecular target drugs (2015)
  • Author(s): Yusuke Ohba et al.
  • Organizer: International Symposium on Multi-dimensional Fluorescence Live Imaging of Cellular Function and Molecular Activities
  • Place of Presentation: 国立京都国際会館 （京都府京都市）
  • Year and Date: 2015-01-26 – 2015-01-27
• [Presentation] Ca2+ signaling mediated influenza virus internalization into host cells (2015)
  • Author(s): Yoichiro Fujioka et al.
  • Organizer: International Symposium on Multi-dimensional Fluorescence Live Imaging of Cellular Function and Molecular Activities
  • Place of Presentation: 国立京都国際会館 （京都府京都市）
  • Year and Date: 2015-01-26 – 2015-01-27
• [Presentation] Ca2+ signaling is involved in influenza virus entry into host cells (2014)
  • Author(s): Yoichiro Fujioka et al.
  • Organizer: The 2014 ASCB/IFCB meeting
  • Place of Presentation: Pennsylvania Convention Center (Philadelphia, PA, USA)
  • Year and Date: 2014-12-07
• [Presentation] FRETバイオセンサーを用いたCML生細胞におけるBCR-ABL活性測定 (2014)
  • Author(s): 藤岡真理他
  • Organizer: 第37回日本分子生物学会
  • Place of Presentation: パシフィコ横浜（神奈川県 横浜市）
  • Year and Date: 2014-11-27
• [Presentation] Ras-PI3Kによるエンドサイトーシス制御に関与する因子の機能解析 (2014)
  • Author(s): 堀内浩水他
  • Organizer: 第37回日本分子生物学会
  • Place of Presentation: パシフィコ横浜（神奈川県 横浜市）
  • Year and Date: 2014-11-27
• [Presentation] Ras-PI3Kシグナルによるエンドサイトーシス制御に関与する分子の探索 (2014)
  • Author(s): 佐藤 絢他
  • Organizer: 第37回日本分子生物学会
  • Place of Presentation: パシフィコ横浜（神奈川県 横浜市）
  • Year and Date: 2014-11-27
• [Presentation] Ca2+シグナルを介したインフルエンザウイルス宿主細胞侵入メカニズムの解明 (2014)
  • Author(s): 藤岡容一朗他
  • Organizer: 第37回日本分子生物学会
  • Place of Presentation: パシフィコ横浜（神奈川県 横浜市）
  • Year and Date: 2014-11-26
• [Presentation] 光による細胞の『観察』から『操作』へ (2014)
  • Author(s): 大場雄介
  • Organizer: 第37回日本分子生物学会
  • Place of Presentation: パシフィコ横浜（神奈川県 横浜市）
  • Year and Date: 2014-11-25
  • Invited
• [Presentation] イメージング技術を用いた分子標的治療薬効果判定法の開発と応用 (2014)
  • Author(s): 大場雄介
  • Organizer: 第4回北海道探索病理学研究シンポジウム
  • Place of Presentation: 京王プラザホテル札幌（北海道札幌市）
  • Year and Date: 2014-10-25
  • Invited
• [Presentation] 癌細胞におけるシグナル伝達イメージングとその応用 (2014)
  • Author(s): 大場雄介
  • Organizer: 第87回日本生化学会大会
  • Place of Presentation: 国立京都国際会館（京都府京都市）
  • Year and Date: 2014-10-17
  • Invited
• [Presentation] ウイルス-宿主相互作用を可視化する (2014)
  • Author(s): 南保明日香他
  • Organizer: 第87回日本生化学会大会
  • Place of Presentation: 国立京都国際会館（京都府京都市）
  • Year and Date: 2014-10-15
  • Invited
• [Presentation] Clinical application of fluorescence imaging (2014)
  • Author(s): 大場雄介
  • Organizer: 第73回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜（神奈川横浜市）
  • Year and Date: 2014-09-27
  • Invited
• [Presentation] カルシウムシグナルを介した外来因子の細胞取り込み機構 (2014)
  • Author(s): 藤岡容一朗他
  • Organizer: 第94回北海道医学大会生理系分科会
  • Place of Presentation: 北海道大学学術交流会館（北海道札幌市）
  • Year and Date: 2014-08-30
• [Presentation] Ras-PI3Kシグナルによるエンドサイトーシス制御機構の解明 (2014)
  • Author(s): 佐藤絢他
  • Organizer: 第94回北海道医学大会生理系分科会
  • Place of Presentation: 北海道大学学術交流会館（北海道札幌市）
  • Year and Date: 2014-08-30
• [Presentation] インフルエンザウイルスのCa+2シグナルを介した宿主細胞侵入機構 (2014)
  • Author(s): 藤岡容一朗他
  • Organizer: 第66回日本細胞生物学会大会
  • Place of Presentation: 奈良県新公会堂（奈良県奈良市）
  • Year and Date: 2014-06-11
• [Presentation] フェルスター共鳴エネルギー移動を用いたシグナル伝達の可視化とその応用 (2014)
  • Author(s): 大場雄介
  • Organizer: 第91回日本生理学会大会
  • Place of Presentation: 鹿児島大学郡元キャンパス（鹿児島）
  • Invited
• [Presentation] Involvement of Ras-PI3K-mediated calcium signaling in the regulation of endocytosis (2014)
  • Author(s): Y. Fujioka, M. Tsuda, A. Nanbo, T. Hattori, J. Sasaki, T. Sasaki, T. Miyazaki and Y. Ohba
  • Organizer: 第91回日本生理学会大会
  • Place of Presentation: 鹿児島大学郡元キャンパス（鹿児島）
• [Presentation] Influenza viruses internalize into host cells via calcium signaling (2013)
  • Author(s): Y. Fujioka, M. Tsuda, A. Nanbo, T. Hattori, J. Sasaki, T. Sasaki, T. Miyazaki and Y. Ohba
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 神戸国際会議場他（神戸）
• [Presentation] P-gp阻害剤はメトロノミックケモセラピーの血管新生阻害効果を強める (2013)
  • Author(s): 秋山廣輔，大賀則孝，樋田泰浩，間石奈湖，Alam Mohammad Towfik，川本泰輔，大村瞳，山田健司，鳥居ちさほ，進藤正信，大場雄介，樋田京子
  • Organizer: 第72回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜（横浜）
• [Presentation] 血管内皮細胞によるがん転移促進 (2013)
  • Author(s): 間石奈湖，大場雄介，大賀則孝,秋山廣輔,山本和幸，浜田淳一，川本泰輔，Alam Mohammad Towfik, 大村瞳，進藤正信，樋田泰浩，樋田京子
  • Organizer: 第72回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜（横浜）
• [Presentation] 細胞機能の蛍光バイオイメージング (2013)
  • Author(s): 大場雄介
  • Organizer: 第33回日本骨形態計測学会
  • Place of Presentation: アクトシティ浜松コングレスセンター（浜松）
  • Invited
• [Presentation] Involvement of Ras-PI3K signaling in the uptake of exogenous factors into cells (2013)
  • Author(s): Y. Fujioka, M. Tsuda, A. Nanbo, T. Hattori, J. Sasaki, T. Sasaki, T. Miyazaki and Y. Ohba
  • Organizer: 第65回日本細胞生物学会大会
  • Place of Presentation: ウインクあいち（名古屋）
• [Presentation] 細胞機能とシグナル伝達の蛍光イメージング
  • Author(s): 大場雄介
  • Organizer: 日本生化学会北海道支部例会第50回記念大会
  • Place of Presentation: 北海道大学医学部学友会館（札幌市）
  • Invited
• [Book] Fluorescence-Based Biosensors-concepts and applications. Progress in Molecular Biology and Translational Science (2013)
  • Author(s): Y, Ohba, Y. Fujioka, S. Nakada and M. Tsuda
  • Total Pages: 429
  • Publisher: Elsevier, London
• [Patent(Industrial Property Rights)] フェルスター共鳴エネルギー移動用ポリペプチド (2014)
  • Inventor(s): 大場雄介
  • Industrial Property Rights Holder: 大場雄介
  • Industrial Property Rights Type: 特許
  • Filing Date: 2014-04-08
  • Overseas
• [Patent(Industrial Property Rights)] フェルスター共鳴エネルギー移動用ポリペプチド (2013)
  • Inventor(s): 大場雄介
  • Industrial Property Rights Holder: 北海道大学
  • Industrial Property Rights Type: 特許
  • Filing Date: 2013-04-08