キナーゼ阻害剤感受性パネルに基づく白血病の層別化

• Project/Area Number: 20014008
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Nagoya University
• Principal Investigator: 清井 仁 Nagoya University, 医学部附属病院, 講師 (90314004)
• Project Period (FY): 2008 – 2009
• Project Status: Completed (Fiscal Year 2009)
• Budget Amount: ¥10,700,000 (Direct Cost: ¥10,700,000); Fiscal Year 2009: ¥4,700,000 (Direct Cost: ¥4,700,000); Fiscal Year 2008: ¥6,000,000 (Direct Cost: ¥6,000,000)
• Keywords: キナーゼ / 白血病 / 阻害剤 / 分子標的療法 / シグナル伝達

Research Abstract

本研究では、各種キナーゼ阻害剤に対する感受性を基に白血病の層別化を試みた。
昨年度までに、受容体型チロシンキナーゼであるFLT3およびKIT阻害剤の感受性がリガンド刺激によって軽減されることを明らかにしたことより、膜結合型リガンド(FL及びSCF)発現支持細胞を樹立し、これらとの共培養系での評価を追加した。
1. リガンド発現細胞との接着状態での共発現培養系では、選択性の高い阻害剤による増殖抑制効果の減弱が、液体培養系よりもより顕著に認められることが明らかとなった。
2. この阻害効果の減弱は正常キナーゼ分子の発現が高いものほど強い傾向にあることから、正常および変異型FLT3共発現細胞を樹立して検討したところ、正常FLT3分子を介したリガンド依存性シグナルが阻害剤の効果を減弱していることが明らかとなった。
3. 一方、マルチキナーゼ阻害活性を有する化合物では、リガンド刺激による阻害効果の減弱が弱いことから、選択性の高い阻害剤との比較を行う事により、より白血病細胞の依存する活性化キナーゼを同定することが可能であり層別化に有用であることを明らかにした。
4. 上記方法とKIT、FLT3選択的阻害剤を用いることにより、KIT、FLT3キナーゼに依存度の高いAMLが層別化され、これら阻害剤の治療効果予測に有用と考えられた。
5. trans-golgiでの糖鎖修飾と分泌を阻害するmonencinを用いることにより、大多数のAML細胞の増殖および支持細胞との接着が阻害された。これは受容体型キナーゼが細胞表面に発現されないために誘導される現象であり、monencin耐性の細胞では細胞質内での変異型キナーゼの活性化が認められた。
これらの結果から、受容体型チロシンキナーゼ依存性が高く、阻害剤の有効性が期待される一群を抽出した。monencin耐性AML細胞質内で活性化している未知のキナーゼをリン酸化抗体アレイで同定することにより新たな変異キナーゼの同定を試みた。

Research Products

• [Journal Article] Phase I Study of KW-0761, a Defucosylated Humanized Anti-CCR4 Antibody, in Relapsed Patients With Adult T-Cell Leukemia-Lymphoma and Peripheral T-Cell Lymphoma. (2010)
  • Author(s): Kazuhito Yamamoto
  • Journal Title: J Clin Oncol 28 Pages: 1591-1598
  • Peer Reviewed
• [Journal Article] Randomized trial of response-oriented individualized versus fixed-schedule induction chemotherapy with idarubicin and cytarabine in adult acute myeloid leukemia : the JALSG AML95 study. (2010)
  • Author(s): Satoshi Ohtake
  • Journal Title: Int J Hematol 91 Pages: 276-283
  • Peer Reviewed
• [Journal Article] BCR-ABL-independent and RAS/MAPK pathway-dependent form of imatinib resistance in Ph-positive acute lymphoblastic leukemia cell line with activation of EphB4. (2010)
  • Author(s): Momoko Suzuki
  • Journal Title: Eur J Haematol 84 Pages: 229-238
  • Peer Reviewed
• [Journal Article] Donor single nucleotide polymorphism in the CCR9 gene affects the incidence of skin GVHD. (2010)
  • Author(s): Yoshihiro Inamoto
  • Journal Title: Bone Marrow Transplant 45 Pages: 363-369
  • Peer Reviewed
• [Journal Article] Irrespective of CD34 expression, lineage-committed cell fraction reconstitutes and re-establishes transformed Philadelphia chromosome-positive leukemia in NOD/SCID/IL-2Rgammac mice. (2009)
  • Author(s): Ryohei Tanizaki
  • Journal Title: Cancer Sci (in press)
  • Peer Reviewed
• [Journal Article] Escape mechanisms from antibody therapy to lymphoma cells : downregulation of CD20 mRNA by recruitment of the HDAC complex and not by DNA methylation. (2009)
  • Author(s): Takumi Sugimoto
  • Journal Title: Biochem Biophys Res Commun 390 Pages: 48-53
  • Peer Reviewed
• [Journal Article] KW-2449, a novel multi-kinase inhibitor, suppresses the growth of leukemia cells with FLT3 mutations or T315I-mutated BCR/ABL translocation. (2009)
  • Author(s): Yukimasa Shiotsu
  • Journal Title: Blood 114 Pages: 1607-1617
  • 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 lymph ocytic leukaemia and lymphoma. (2009)
  • Author(s): Akihiro Takeshita
  • Journal Title: Br J Haematol 146 Pages: 34-43
  • Peer Reviewed
• [Journal Article] CMC-544 (inotuzumab ozogamicin), an anti-CD22 immuno-conjugate of calicheamicin, alters the levels of target molecules of malignant B-cells. (2009)
  • Author(s): Akihiro Takeshita
  • Journal Title: Leukemia 146 Pages: 34-43
  • Peer Reviewed
• [Journal Article] Comprehensive analysis of cooperative gene mutations between class I and class II in de novo acute myeloid leukemia. (2009)
  • Author(s): Yuichi Ishikawa
  • Journal Title: Eur J Haematol 83 Pages: 90-98
  • Peer Reviewed
• [Journal Article] Down-regulation of CD20 expression in B-cell lymphoma cells after treatment with rituximab-containing combination chemotherapies : its prevalence and clinical significance. (2009)
  • Author(s): Junji Hiraga
  • Journal Title: Blood 113 Pages: 4885-4893
  • Peer Reviewed
• [Journal Article] Abnormal cytoplasmic dyslocalisation and/or reduction of nucleophosmin protein level rarely occurs in myelodysplastic syndromes (2008)
  • Author(s): Yuichi IshikaWa
  • Journal Title: Leuk Lymphoma 49 Pages: 2359-2364
  • Peer Reviewed
• [Journal Article] Retention but significant reduction of BCR-ABL transcript in heraatopoietic stem cells in chronic myelogenous leukemia after imatimib therapy (2008)
  • Author(s): Akihiro Abe
  • Journal Title: Int J Hernatol 88 Pages: 471-475
  • Peer Reviewed
• [Journal Article] Mutational analysis of SOS 1 gene in acute myeloid leukemia (2008)
  • Author(s): Ryohei Tanizaki
  • Journal Title: Int J Hematol 88 Pages: 460-462
  • Peer Reviewed
• [Journal Article] Novel and orally active 5-(1, 3, 4-oxadiazol-2-yl)pyrinaidine deriva tives as selective FLT 3 inhibitors (2008)
  • Author(s): Hiroshi Ishida
  • Journal Title: Bioorg Med Chem Lett 18 Pages: 5472-5477
  • Peer Reviewed
• [Journal Article] Prognostic implication and biological roles of RhoH in acute myeloid leukaemia (2008)
  • Author(s): Toshihiro Iwasaki
  • Journal Title: Eur J Haematol 81 Pages: 454-460
  • Peer Reviewed
• [Journal Article] Karyotypically independent clones with del (11q) and trisomy 10 in acute myeloid leukemia : trisomy 10 may appear as an additional change (2008)
  • Author(s): Moritaka Gotoh
  • Journal Title: Int J Hematol 88 Pages: 123-124
  • Peer Reviewed
• [Journal Article] Clinical significance of minimal residual disease in patients with t(8 ; 21) acute myeloid leukemia in Japan (2008)
  • Author(s): Hiroto Narimatsu
  • Journal Title: Int J Hematol 88 Pages: 154-158
  • Peer Reviewed
• [Journal Article] Donor cell leukemia after allogeneic peripheral blood stem celltransplantation : a case report and literature review (2008)
  • Author(s): Makoto Murata
  • Journal Title: Int J Hematol 88 Pages: 111-115
  • Peer Reviewed
• [Journal Article] Functional analysis of JAK 3 mutations in transient myeloproliferative disorder and acute megakaryoblastic leukaemia accompanying Down syndrome (2008)
  • Author(s): Tomohiko Sato
  • Journal Title: Br J Haematol 141 Pages: 681-688
  • Peer Reviewed
• [Presentation] MS4A1 (CD20) Gene Expression Is Down-Regulated by Recruiting the Histone Deacetylase Protein Complex to the Promoter in the CD20-Negative B-Lymphom a Cells After Treatment with Rituximab. (2009)
  • Author(s): Sugimoto Takumi
  • Organizer: The American Society of Hematology 51th Annual Meeting
  • Place of Presentation: New Orleans USA
• [Presentation] Treatment with Bortezomib Overcomes Resistance to Imatinib in Ph-Leukemia Quiescent Cells. (2009)
  • Author(s): Kuwatsuka Yachiyo
  • Organizer: The American Society of Hematology 51th Annual Meeting
  • Place of Presentation: New Orleans USA
• [Presentation] Double Genetic Mutations in PML-Rara Fusion Gene Confirmed in a Patient Showing Resistance to All-Trans Retinoic Acid and Arsenic-Trioxide Therapy. (2009)
  • Author(s): Goto Emi
  • Organizer: The American Society of Hematology 51th Annual Meeting
  • Place of Presentation: New Orleans USA
• [Presentation] FL-Dependent Wild-Type FLT3 Signals Reduce the Inhibitory Effects of FLT3 Inhibitors On Wild-Type and Mutant FLT3 Co-Expressing Cel. (2009)
  • Author(s): Mori Yumiko
  • Organizer: The American Society of Hematology 51th Annual Meeting
  • Place of Presentation: New Orleans USA
• [Presentation] FLT3/ITD Regulates Leukemia Cell Adhesion through α4β1 Integrin and Pyk2 Signalin. (2009)
  • Author(s): Katsumi Akira
  • Organizer: The American Society of Hematology 51th Annual Meeting
  • Place of Presentation: New Orleans USA
• [Presentation] Treatment with mTOR Inhibitor, Everolimus (RAD001) Overcomes Resistance to Imatinib in Ph-Leukemia Quiescent or T315I-Mutated Cells. (2009)
  • Author(s): Minami Yousuke
  • Organizer: The American Society of Hematology 51th Annual Meeting
  • Place of Presentation: New Orleans USA
• [Presentation] Treatment with Hsp 90 Inhibitor, 17-AAG Overcomes Resistance to Small Molecule FLT 3-Inhibitors in FLT 3/ITD-Positive Leukemia Cells Harboring N 676 K-Mutation (2008)
  • Author(s): Yosuke Minanai
  • Organizer: The American Society of Hematology 50 th Annual Meeting
  • Place of Presentation: San Francisco USA
• [Presentation] A Randomized Trial Comparing Individualized Vs. Non-Individualized Treatment for Elderly Acute Myeloid Leukemia : JALSG GML 200 Study (2008)
  • Author(s): Atsushi Wakita
  • Organizer: The American Society of Hematology 50 th Annual Meeting
  • Place of Presentation: San Francisco USA
• [Presentation] The Genotype Consisting of Complex Karyotype and TP 53 Gene Mutation Is Specific to Acute Myeloid Leukemia with Multilineage Dysplasia (2008)
  • Author(s): Yuichi Ishikawa
  • Organizer: The American Society of Hematology 50 th Annual Meeting
  • Place of Presentation: San Francisco USA
• [Presentation] Prognostic Implication and Biological Roles of RhoH in Acute Myeloid Leukemia (2008)
  • Author(s): Akira Katsumi
  • Organizer: The American Society of Hematology 50th Annual Meeting
  • Place of Presentation: San Francisco USA
• [Presentation] KW-2449, a Novel Multi-Kinase Inhibitor, Suppresses the Growth of Imatinib-Resistant Ph + Leukemia Including BCR-ABL/T 315 I Both in Vitro and in Vivo (2008)
  • Author(s): Yukimasa Shiotsu
  • Organizer: The American Society of Hematology 50 th Annual Meeting
  • Place of Presentation: San Francisco USA
• [Presentation] Updated Results of JALSG AML201 Study Comparing Intensified Daunorubicin with Idarubicin in Patients with De Novo Acute Myeloid Leukemia : Effect of Hematopoietic Stem Cell Transplantation (2008)
  • Author(s): Shigeki Ohtake
  • Organizer: The American Society of Hematology 50th Annual Meeting
  • Place of Presentation: San Francisco USA
• [Presentation] Long-Term Follow-up of the Randomized JALSG AML 201 Study Comparing High Dose Ara-C Therapy with Conventional Consolidation Therapy in Adult Acute Myeloid Leukemia (AML) (2008)
  • Author(s): Shuichi Miyawaki
  • Organizer: The American Society of Hematology 50th Annual Meeting
  • Place of Presentation: San Francisco USA
• [Presentation] Relationship Between Post-Translational Modification of CD 20 Protein and the Responsiveness to Rituximab Treatment (2008)
  • Author(s): Taknini Sugimoto
  • Organizer: The American Society of Hematology 50 th Annual Meeting
  • Place of Presentation: San Francisco USA
• [Presentation] Retention but Significant Reduction of BCR-ABL Transcript in Hematopoietic Stem Cells in Chronic Myelogenous Leukemia after Imatinib Therapy (2008)
  • Author(s): Akihiro Abe
  • Organizer: The American Society of Hematology 50 th Annual Meeting
  • Place of Presentation: San Francisco USA