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Identification of minimal residual cells after the treatment of leukemia cells by the imaging of mRNA

Research Project

Project/Area Number 19590552
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Laboratory medicine
Research InstitutionHamamatsu University School of Medicine

Principal Investigator

TAKESHITA Akihiro  Hamamatsu University School of Medicine, 医学部, 准教授 (00242769)

Co-Investigator(Kenkyū-buntansha) TAKESHITA Kaori  浜松医科大学, 医学部附属病院, 医員 (30397393)
MAEKAWA Masato  浜松医科大学, 医学部, 教授 (20190291)
Project Period (FY) 2007 – 2008
Project Status Completed (Fiscal Year 2008)
Budget Amount *help
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2008: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2007: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Keywords白血病 / 微少残存病変 / mRNA / flow cytometry / 白血病細胞 / Wt-1 / 生細胞
Research Abstract

特定の白血病細胞の分画の検出と分離を施行するため、細胞内のmRNAの染色を試みた。QUAL/FRET(quenched autoligating/fluorescence resonance energy transfer)法を改良し、蛍光probeの導入技術や反応時間の調整など多方面の改良を施行した。適切なdonor probe とacceptor probeを設定し、energy transferにて得られた特異的蛍光を検出しようとした。標的分子(mRNA)をWT1とし、白血病細胞の同定と分離を生細胞の状態で行う可能性を模索した。作成されたprobeの細胞膜の透過性を一時的に高めるためstreptlysin O(SLO)を使用した。SLOの至適濃度の決定に関しては細胞や試薬の状態などによる再現性の低下を防止するため、probe導入前にSLOの処理濃度の最適化実験を行い、flow cytometryにて検討した。培養細胞では細胞内蛍光の増加を認め、WT1が特異的に検出できると考えられた。しかし、WT1の発現が本来非常に弱いとされている正常のリンパ球における蛍光がprobe量の増加とともに増加し、non-specificな蛍光をより低下させる必要性がでてきた。Non-specificな蛍光の原因を検討するため、分子量を変えたprobeを設計して細胞内に導入した。またprobeが核内へ移行することも考えられたため、probeに接着させる蛍光物質の量を変化させ検討している。

Report

(3 results)
  • 2008 Annual Research Report   Final Research Report ( PDF )
  • 2007 Annual Research Report
  • Research Products

    (31 results)

All 2009 2008 2007 Other

All Journal Article (18 results) (of which Peer Reviewed: 11 results) Presentation (8 results) Book (5 results)

  • [Journal Article] Effective blood utilization via system for massive blood transfusion, including cardiovascular operation in local areas2009

    • Author(s)
      Takeshita A, Asai T, Murakami M, Fujihara H, Ishizuka T, Nakai S, Yamada C, Suzumura T, Uchiyama Y, Maekawa M, Shigeno K, Washiyama N, Yamashita K, Unno N, Shinjo K
    • Journal Title

      Jpn J Transfusion and Cell Therapy 55

      Pages: 64-67

    • NAID

      10025628900

    • Related Report
      2008 Final Research Report
  • [Journal Article] Effective blood utilization via system for massive blood trans fusion, including cardiovascular operation in local areas2009

    • Author(s)
      Takeshita A, Asai T, Murakami M, Fujihara H, Ishizuka T, Nakai S, Yamada C, Suzumura T, Uchiyama Y, Maekawa M, Shigeno K, Washiyama N, Yamashita K, Unno N, Shinjo K.
    • Journal Title

      Jap J Transfusion and Cell Therapy 55

      Pages: 64-67

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Phase I/II study of humanized anti-CD33 antibody conjugated with calicheamicin, gemtuzumab ozogamicin, in relapsed or refractory acute my eloid leukemia : Final results of Japanese multi-center cooperative study2009

    • Author(s)
      Kobayashi Y, Naito K, Takeshita A, Morishima Y, Ogura M, Besso H, Hotta T, Mitani K, Takeuchi H, Miyawaki S, Naoe T, Ohno R
    • Journal Title

      Int J Hematol (in press)

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] CMC544 (inotuzumab ocogamicin), an ati-CD22 immuno-conjugate of calicheamicin, alters the levels of target molecules of malignant B- cells2009

    • Author(s)
      Takeshita A, Yamakage N, Shinjo K, Ono T, Hirano I, Nakamura S, Shigeno K, Tobita T, Maekawa M, Kiyoi H, Naoe T, Ohnishi K, Sugimoto Y, Ohno R
    • Journal Title

      Leukemia (in press)

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] CMC-544 (inotuzumab ozogamicin) shows less effect on multidrug resistant cells : analyses in cell lines and cells from patients with B-cell chronic lymphocytic leukemia and lymphoma2009

    • Author(s)
      Takeshita A, Shinjo K, Yamakage N, Ono T, Hirano I, Matsui H, Nakamura S, Shigeno K, Tobita T, Maekawa M, Kiyoi H, Naoe T, Ohnishi K, Sugimoto Y, Ohno R
    • Journal Title

      Br J Hematol (in press)

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Pharmacokinetics of arsenic species in Japanese patients with relapsed or refractory acute promyelocytic leukemia treated with arsenic trioxide2007

    • Author(s)
      Fujisawa S, Ohno R, Shigeno K, Sahara N, Nakamura S, Naito K, Kobayashi M, Shinjo K, Takeshita A, Suzuki Y, Hashimoto H, Kinoshita K, Shimoya M, Kaise T, Ohnishi K
    • Journal Title

      Cancer Chemother Pharmacol 54

      Pages: 485-493

    • Related Report
      2008 Final Research Report
  • [Journal Article] A randomized study with or without intensified maintenance chemotherapy in patients with acute promyelocytic leukemia who have become negative for PML-RARα transcript after consolidation therapy : The Japan Adult Leukemia Study Group(JALSG)APL97 study2007

    • Author(s)
      Asou N, KIshimoto Y, Kiyoi H, Okada M, Kawai Y, Tsuzuki M, Horikawa K, Matsuda M, Shinagawa K, Ohtake S, NishimuraM, Takahashi M, Yagasaki F, Takeshita A, Kimura Y, Iwanaga M, Naoe T, Ohno R
    • Journal Title

      Blood 110

      Pages: 59-66

    • Related Report
      2008 Final Research Report
  • [Journal Article] Phase I Trial of FLAGM with High Doses of Cytosine Arabinoside for Relapsed, Refractory Acute Myeloid Leukemia : Study of the Japan Adult Leukemia Study Group(JALSG)2007

    • Author(s)
      Miyawaki S, Kawai Y, Takeshita A, Komatsu N, Usui N, Arai Y, Ishida F, Morii T, Kano Y, Ogura M, Doki N, Ohno R
    • Journal Title

      Int J Heamatol 86

      Pages: 343-347

    • NAID

      10019990223

    • Related Report
      2008 Final Research Report
  • [Journal Article] Pharmacokinetics of arsenic species in Japanese patients with relapsed or refractoryacute promyelocytic leukemia treated with arsenic trioxide2007

    • Author(s)
      Fujisawa S, Ohno R, Shigeno K, Sahara N, Nakamura S, Naito K, Kobayashi M, Shinjo K, Takeshita A, Suzuki Y, Hashimoto H, Kinoshita K, Shimoya M, Kaise T, Ohnishi K.
    • Journal Title

      Cancer Chemother Pharmacol 59(4)

      Pages: 485-493

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A randomized study with or without intensified maintenance chemotherapy in patients with acute promyelocytic leukemia who have become negative for PML-RARa transcript after consolidation therapy:The Japan Adult Leukemia Study Group(JALSG)APL97 study2007

    • Author(s)
      Asou N, KIshimoto Y, Kiyoi H, Okada M, Kawai Y, Tsuzuki M, Horikawa K, Matsuda M, Shinagawa K, Ohtake S, Nishimura M, Takahashi M, Yagasaki F, Takeshita A, Kimura Y, Iwanaga M, Naoe T, Ohno R
    • Journal Title

      Blood 110(1)

      Pages: 59-66

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Phase I Trial of FLAGM with High Doses of Cytosine Arabinoside for Relapsed, Refractory Acute Myeloid Leukemia:Study of the Japan Adult Leukemia Study Group (JALSG)2007

    • Author(s)
      Miyawaki S, Kawai Y, Takeslvta A, Komatsu N, Usui N, Arai Y, Ishida F, Morii T, Kano Y, Ogura M, Doki N, Ohno R
    • Journal Title

      Int J Hematol 86

      Pages: 343-347

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Clinical Significance of P-Glycoprotein in Acute Leukemia and a Strategy to Overcome Drug Resistance2007

    • Author(s)
      Motoji T, Motomura S, Wang Y, .Tsuji K, Takanashi M, Slvozaki H, Mlyawaki S, Asou N, Takeshita A, Saburi Y, Ohno R, Mizoguchi H
    • Journal Title

      Frontiers in Cancer Res

      Pages: 123-151

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] 造血器腫瘍の分子標的療法。標準的治療にみる分子標的療法レチノイン酸、Am80による急性前骨髄球性白血病の治療急性前骨髄球性白1(ll病に対する全トランス型レチノイン酸と合成レチノイド.Am80の標準的な治療と発展性2007

    • Author(s)
      竹下 明裕
    • Journal Title

      医学のあゆみ 220(9)

      Pages: 681-686

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] 造血器腫瘍基礎・臨床領域における最新の研究動向。臨床編治療法抗体療法白血病の抗体療法2007

    • Author(s)
      竹下 明裕
    • Journal Title

      日本臨床 65(1)

      Pages: 408-415

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] 造血器腫瘍基礎・臨床領域における最新の研究動向。臨床編診断法細胞表面マーカー(解説/特集〉2007

    • Author(s)
      竹下 明裕
    • Journal Title

      日本臨床 65(1)

      Pages: 293-298

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Phase I/II study of humanized anti-CD33 antibody conjugated with calicheamicin, gemtuzumab ozogamicin, in relapsed or refractory acute myeloid leukemia : Final results of Japanese multi-center cooperative study

    • Author(s)
      Kobayashi Y, Naito K, Takeshita A, Morishima Y, Ogura M, Besso H, Hotta T, Mitani K, Takeuchi H, Miyawaki S, Naoe T, Ohno R.
    • Journal Title

      Int J Hematol (in press)

    • Related Report
      2008 Final Research Report
  • [Journal Article] CMC544(inotuzumab ocogamicin), an ati-CD22 immuno-conjugate of calicheamicin, alters the levels of target molecules of malignant B-cells

    • Author(s)
      Takeshita A, Yamakage N, Shinjo K, Ono T, Hirano I, Nakamura S, Shigeno K, Tobita T, Maekawa M, Kiyoi H, Naoe T, Ohnishi K, Sugimoto Y, Ohno R
    • Journal Title

      Leukemia (in press)

    • Related Report
      2008 Final Research Report
  • [Journal Article] CMC-544(inotuzumab ozogamicin)shows less effect on multidrug resistant cells : analyses in cell lines and cells from patients with B-cell chronic lymphocytic leukemia and lymphoma

    • Author(s)
      Takeshita A, Shinjo K, Yamakage N, Ono T, Hirano I, Matsui H, Nakamura S, Shigeno K, Tobita T, Maekawa M, Kiyoi H, Naoe T, Ohnishi K, Sugimoto Y, Ohno R
    • Journal Title

      Br J Haematol (in press)

    • Related Report
      2008 Final Research Report
  • [Presentation] A Randomized Trial Comparing Individualized Vs. Non-Individualized Treatment for Elderly Acute Myeloid Leukemia : JALSG GML200 study2008

    • Author(s)
      Wakita A, Ohtake S, Takada S, Yagasaki F, Komatsu H, Miya zaki Y, Kubo K, Kimura Y, Takeshita A, Adachi Y, Kiyoi H, Yamaguchi T, Yoshida M, Ohnishi K, Miyawaki S, Naoe T, Ueda R, Ohno R
    • Organizer
      American Society of Hematology
    • Place of Presentation
      Sanfrancisco, USA
    • Year and Date
      2008-10-05
    • Related Report
      2008 Annual Research Report
  • [Presentation] The usefulness of the expected elevation value of serum albumin before replacement therapy2008

    • Author(s)
      Fujihara H, Takeshita K, Nakai S, Yamada C, Suzumura T, Uchiyama S, Takeshita A
    • Organizer
      American Association of Blood Banking
    • Place of Presentation
      Montreal, Canada
    • Year and Date
      2008-05-10
    • Related Report
      2008 Annual Research Report
  • [Presentation] Cell cycle features and quantitative alteration of target molecules of malignant B cells treated with CMC544 alone or in combination with rituximab2007

    • Author(s)
      Yamakage N, Takeshita A, Shinjo K, Ono T, Hiranol, Okinaka K, Nakamura S, Shigeno K, Maekawa M, Ohnishi K, Ohno R
    • Organizer
      American Society of Hematology
    • Place of Presentation
      アトランタ(米国)
    • Year and Date
      2007-12-09
    • Related Report
      2007 Annual Research Report
  • [Presentation] Reduced effect of CMC544 on P-glycoprotein positive malignant B cells and its restoration by multidrug resistance modifiers2007

    • Author(s)
      Takeshita A, Shinjo K, Yamakage N Ono T, Hiranol, Okinaka K, Matsui H, Nakamura S, Shigeno K, Maekawa M, Ohnishi K, Sugimoto Y, Ohno R
    • Organizer
      American Society of Hematology
    • Place of Presentation
      アトランタ(米国)
    • Year and Date
      2007-12-09
    • Related Report
      2007 Annual Research Report
  • [Presentation] Cell cycle features and quantitative alteration of target molecules of malignant B cells treated with CMC544 alone or in combination with rituximab2007

    • Author(s)
      Yamakage N, Takeshita A, ShinjoK, Ono T, Hirano I, Okinaka K, Nakamura S, Shigeno K, Maekawa M, Ohnishi K, Ohno R
    • Organizer
      Annual Meeting of American Society of Hematology, Sanfrancisco, USA
    • Place of Presentation
      Atranta, USA, Dec
    • Related Report
      2008 Final Research Report
  • [Presentation] Reduc ed effect of CMC544 on P-glycoprotein po sitive malignant B cells and its restoration by multidrug resistance modifiers2007

    • Author(s)
      Takeshita A, Shinjo K, YamakageN, O no T, Hirano I, Okinaka K, Matsui H, Nakamura S, Shigeno K, Maekawa M, Ohnishi K , Sugimoto Y , Ohno R
    • Organizer
      Annual Meeting of American Society of Hematology, Sanfrancisco, USA
    • Place of Presentation
      Atranta, USA, Dec
    • Related Report
      2008 Final Research Report
  • [Presentation] The usefulness of the expected elevation value of serum albumin before replacement therapy

    • Author(s)
      Fujihara H, Takeshita K, Nakai S, Yamada C, Suzumura T, Uchiyama S, Takeshita A.
    • Organizer
      Annual Meeting of AABB Montreal
    • Place of Presentation
      Canada
    • Related Report
      2008 Final Research Report
  • [Presentation] A Randomized Trial Comparing Individualized Vs. Non-Individualized Treatment for Elderly Acute Myeloid Leukemia : JALSG GML200 Study

    • Author(s)
      Wakita A, Ohtake S, Takada S, Yagasaki F, Komatsu H, Miyazaki Y, Kubo K, Kimura Y, Takeshita A, Adachi Y, Kiyoi H, Yamaguchi T, Yoshida M, Ohnishi K, Miyawaki S, Naoe T, Ueda R, Ohno R.
    • Organizer
      Annual Meeting of American Society of Hematology
    • Place of Presentation
      Sanfrancisco, USA
    • Related Report
      2008 Final Research Report
  • [Book] 腫瘍内科2009

    • Author(s)
      竹下明裕, 直江知樹
    • Publisher
      科学評論社
    • Related Report
      2008 Annual Research Report
  • [Book] Annual review2009

    • Author(s)
      竹下明裕
    • Publisher
      中外医学社
    • Related Report
      2008 Annual Research Report
  • [Book] medicina2008

    • Author(s)
      竹下明裕
    • Publisher
      医学書院
    • Related Report
      2008 Annual Research Report
  • [Book] 周産期医学2008

    • Author(s)
      竹下明裕
    • Publisher
      東京医学社
    • Related Report
      2008 Annual Research Report
  • [Book] Clinical Significance of P-Glycoprotein in Acute Leukemia and a Strategy to Overcome Drug Resistance

    • Author(s)
      Motoji T, Motomura S, Wang Y, Tsuji K, Takanashi M, Shiozaki H, Miyawaki S, Asou N, Takeshita A, Saburi Y, Ohno R, Mizoguchi H
    • Publisher
      Frontiers in Cancer Res
    • Related Report
      2008 Final Research Report

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Published: 2007-04-01   Modified: 2016-04-21  

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