Eradication of chronic myelogenous leukemia stem cells and abnormal clone with T315I point mutation

2011 Fiscal Year Final Research Report

• Project/Area Number: 21390293
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Hematology
• Research Institution: Kyoto University
• Principal Investigator: MAEKAWA Taira 京都大学, 医学研究科, 教授 (80229286)
• Co-Investigator(Kenkyū-buntansha): ASHIHARA Eishi 京都府立医科大学, 医学研究科, 講師 (70275197) ; KIMURA Shinya 佐賀大学, 医学部, 教授 (80359794) ; HIRAI Hideyo 京都大学, 医学研究科, 助教 (50315933) ; MIURA Yasuo 京都大学, 医学研究科, 助教 (70605146)
• Project Period (FY): 2009 – 2011
• Keywords: 慢性骨髄性白血病 / 幹細胞 / 治療抵抗性 / 分子標的治療

Research Abstract

Imatinib mesylate(IM, GleevecR), which specifically inhibits the autophosphorylation of the Abl tyrosine kinase(TK), has dramatically improved the treatment of chronic myelogenous leukemia(CML). However, resistance is often reported in patients with advanced-stage disease. Several novel TK inhibitors(TKI) of second generation such as dasatinib(SprycelR), nilotinib(TasignaR), and INNO-406(Bafetinib) have been developed that override IM resistance mechanisms caused by point mutations within the Abl kinase domain. Because of their strong affinities for the ATP-binding site compared to IM, these new TKIs may be effective in IM-resistant patients. However, they could not inhibit the phosphorylation of T315I Bcr-Abl harboring the mutation of threonine 315 to isoleucine. We describe the in vitro and in vivo effects of AT9283(1-cyclopropyl-3[5-morpholin-4yl methyl-1H-benzomidazol-2-yl]-urea), a potent inhibitor of kinases Aurora A and Aurora B. AT9283 showed potent antiproliferative activity o n cells transformed by BCR-ABL/T315I. CMLstem cells also survived in the bone marrow niche under severe hypoxia condition. We generated two hypoxia-adapted(HA)-CML cell lines. These HA-CML cells exhibited stem cell-like characteristics. Compared with the respective parental cells, HA-CML cells had higher levels of protein and higher enzyme activity of glyoxalase-I(Glo-I), that detoxifies methylglyoxal, a cytotoxic by-product of glycolysis. In contrast to Abl TKIs, Glo-I inhibitors were much more effective in killing HA-CML cells both in vitro and in vivo. Ii is likely that leukemic cells are able to survive and proliferate in severely hypoxic environments. The hypoxic conditions allow leukemic cells that reside there to become insensitive to cell death stimuli. To target leukemic cells in hypoxic conditions, we focused on the hypoxia-selective cytotoxin, Rakicidin A. produced by the Micromonospora strain. We describe Rakicidin A-induced cell death in HA-CML cells with stem cell-like characteristics. Rakicidin A is a promising compound for targeting TKI-resistant quiescent CML stem cells in the hypoxic environment.

Research Products

• [Journal Article] Galectin-3 induced by leukemia microenvironment promotes drug resistance and bone marrow lodgment in chronic myelogenous leukemia (2011)
  • Author(s): Yamamoto-Sugitani, M., Kuroda, J., Ashihara, E., Nagoshi, H., Kobayashi, T., Matsumoto, Y., Sasaki, N., Kiyota, M., Nakayama, R., Akaji, K., Taki, T., Uoshima, N., Kobayashi, Y., Horiike, S., Maekawa, T.
  • Journal Title: Proc. Natl. Acad. Sci. Volume: 108(42) Pages: 17468-17473
• [Journal Article] Cross-priming of CD8(+) T cells pulsed with autologous apoptotic leukemic cells in immunotherapy 4for elderly patients with acute myeloid leukemia (2011)
  • Author(s): Kitawaki, T., Kadowaki, T., Fukunaga, K., Kasai, Y., Maekawa, T., Ohmori, K., Ito, T., Shimizu, A., Kuzushima, K., Kondo, T.
  • Journal Title: Exp Hematol Volume: 39(4) Pages: 424-433
• [Journal Article] Rakicidin A effectively induces apoptosis in hypoxia adapted Bcr-Abl positive leukemic cells (2011)
  • Author(s): Takeuchi, M., Ashihara, E., Yamazaki, Y., Kimura, S., Nakagawa, Y., Tanaka, R., Yao, H., Nagao, R., Hayashi, Y., Hirai, H.
  • Journal Title: Cancer Sci. Volume: 102(3) Pages: 591-596
• [Journal Article] A phase I/II study of immunotherapy for elderly patients with acute myeloid leukemia using dendritic cells pulsed with autologous apoptotic leukemic cells (2011)
  • Author(s): Kitawaki, T., Kadowaki, T., Fukunaga, K., Kasai, Y., Maekawa, T., Ohmori, K., Itoh, T., Tada, H., Teramukai, S., Shimizu, A., Fukushima, M., Kuzushima, K., Kondo, T.
  • Journal Title: Brit J Haematol Volume: 53(6) Pages: 796-799
• [Journal Article] Combined effects of novel heat shock protein 90 inhibitor NVP-AUY922 and nilotinib in a random mutagenesis screen (2011)
  • Author(s): Tauchi, T., Okabe, S., Ashihara, E., Kimura, S., Maekawa, T.
  • Journal Title: Oncogene Volume: 30(24) Pages: 2789-2797
• [Journal Article] Targeting the Wnt/υ-catenin signaling pathway in human cancers. Expert Opin Ther Targets (2011)
  • Author(s): Yao, H., Ashihara, E.
  • Journal Title: Expert Opin Ther Targets Volume: 15(7) Pages: 873-887
• [Journal Article] Growth inhibition of imatinib-resistant CML cells with the T315 mutation and hypoxia-adaptation by AV65 。Va novel Wnt/υ-catenin signaling inhibitor (2011)
  • Author(s): Nagao, R., Ashihara, E., Kimura, S., Strovel, J. W., Yao, H., Takeuchi, M., Tanaka, R., Hayashi, Y., Hirai, H., Padia, J., Strand, K.
  • Journal Title: Cancer Letter Volume: 312(1) Pages: 91-100
  • DOI: doi:10.1016/j.canlet.2011.08.002
• [Journal Article] Rapid automated detection of ABL kinase domain mutations in imatinib-resistant patients (2011)
  • Author(s): Tanaka, R., Kimura, S., Ashihara, E., Yoshimura, M., Takahashi, N., Wakita, H., Itoh, K., Nishiwaki, K., Suzuki, K., Nagao, R., Yao, H., Hayashi, Y., Satake, S., Hirai, H., Sawada, K-I., Ottmann, O. G., Melo, J. V.
  • Journal Title: Cancer Letter Volume: 312(2) Pages: 228-34
  • DOI: doi:10.1016/j.canlet.2011.08.009
• [Journal Article] A novel Wnt/β-catenin signal inhibitor successfully suppresses progression of multiple myeloma in a mouse model (2011)
  • Author(s): Yao, H., Ashihara, E., Strovel, J. W., Nakagawa, Y., Kuroda, J., Nagao, R., Tanaka, R., Yokota, A., Takeuchi, M., Sakai, K., Shimazaki, C., Taniwaki, M., Strand, K., Padia, J., Hirai, H., Kimura, S.
  • Journal Title: Blood Cancer J Volume: 1
• [Journal Article] Osteoclasts are involved in the maintenance of dormant leukemic cells (2010)
  • Author(s): Yokota, A., Kimura, S., Tanaka, R., Takeuchi, M., Yao, H., Sakai, K., Nagao, R., Kuroda, J., Kamitsuji, Y., Kawata, E.
  • Journal Title: Leuk Res. Volume: 34 Pages: 793-799
• [Journal Article] Future prospect of RNA interference for cancer therapies. Curr Drug Targets. (2010)
  • Author(s): Ashihara, E., Kawata, E.
  • Journal Title: Curr Drug Targets. Volume: 11(3) Pages: 345-360
• [Journal Article] Glyoxalase-I is a novel target against Bcr-Abl+ leukemic cells acquiring stem-like 5characteristics in a hypoxic environment (2010)
  • Author(s): Takeuchi, M., Kimura, S., Kuroda, J., Ashihara, E., Kawatani, M., Osada, H., Umezawa, K., Yasui, E., Imoto, M., Tsuruo, T., Yokota, A., Tanaka, T., Nagao, R., Nakahata, T., Fujiyama, Y.
  • Journal Title: Cell Death Diff Volume: 7 Pages: 1211-1220
• [Journal Article] Activity of the multi-targeted kinase inhibitor, AT9283, in imatinib-resistant BCR-ABL positive leukemic cells (2010)
  • Author(s): Tanaka, R., Squires, M. S., Kimura, S., Yokota, A., Mallet, K., Smyth, T., Thompson, N. T., Nagao, R., Yamauchi, T., Ashihara, E., Ottmann, O. G., Lyons, J. F.
  • Journal Title: Blood Volume: 116(12) Pages: 2089-2095
• [Journal Article] Dual BCR-ABL/LYN tyrosine kinase inhibitor (2009)
  • Author(s): Takeuchi M, Kimura S, Ashihara E, Maekawa T
  • Journal Title: Drug Fut Volume: 34(4) Pages: 261-269
• [Presentation] BCR/ABL-mediated myeloid expansion is promoted by C/EBPβ (2011)
  • Author(s): Hayashi Y, Hirai H, Yao H, Yoshioka S, Satake S, Kamio N, Miura Y, Ashihara E, Fujiyama Y, Tenen DG, Maekawa T
  • Organizer: a regulator of emergency granulopoiesis. 53rd ASH Annual Meeting and Exposition
  • Place of Presentation: San Diego, USA
  • Year and Date: 20111211-13
• [Presentation] 転写因子C/EBPβの慢性骨髄性白血病の病態形成への関与 (2011)
  • Author(s): 平位秀世、林嘉宏、前川平
  • Organizer: 難治性白血病に対する標準的治療法の確立(小林班)平成23年度第2回合同班会議
  • Place of Presentation: 名古屋大学(名古屋)
  • Year and Date: 2011-12-17
• [Presentation] アカデミアの立場から-創薬開発を経験して-.ワークショップ1「TRにおける産学連携の模索」 (2011)
  • Author(s): 前川平
  • Organizer: 第7回トランスレーショナルリサーチワークショップ(TRワークショップ2011)
  • Place of Presentation: 東京
  • Year and Date: 2011-01-28
• [Presentation] Development of new agents in the treatment of imatinib-resistant Ph+ leukemias (2010)
  • Author(s): Maekawa T
  • Organizer: The 30th Internationl Symposium by Princess Takamatsu Foundation for Cancer Research
  • Place of Presentation: Kyoto, Japan
  • Year and Date: 2010-12-23
• [Presentation] T315I遺伝子変異を持つCMLに対するオーロラキナーゼ阻害剤、AT9283の開発について (2010)
  • Author(s): 木村晋也、前川平
  • Organizer: 平成22年度第1回合同班会議
  • Place of Presentation: 名古屋
  • Year and Date: 2010-06-19
• [Presentation] A Novel Automated Assay for the Detection of BCR-ABL Kinase Domain Mutations (2010)
  • Author(s): Tanaka R, Kimura S, Hosomi T, Hirai M, Nagao R, Takeuchi M, Yao H, Sakai K, HiraiH, Ashihara E, Maekawa T
  • Organizer: 15th Congress of the European Hematology Association(EHA)
  • Place of Presentation: Barcelona, Spain
  • Year and Date: 2010-06-12
• [Presentation] Hypoxia-Adapted CML cells are more primitive population and are Eradication by Glyoxalase-1 Inhibitors (2009)
  • Author(s): Takeuchi M, Kimura S, Kuroda J, Ashihara E, Kawatani M, Osada H, Umezawa K, Yasui E, ImotoM, Tsuruo T, Yokota A, Tanaka R, Nagao R, Nakahata T, Fujiyama Y, MaekawaT
  • Organizer: The American Society of Hematology 51st Annual Meeting and Exposition(ASH)
  • Place of Presentation: Ernest N. Morial Convention Center, New Orleans, USA
  • Year and Date: 2009-12-06
• [Presentation] Automated and Rapid Detection of BCR-ABL Kinase Domain Mutations in IM Resistant Patients with Ph+ Leukemias (2009)
  • Author(s): Tanaka R, Kimura S, Hosomi T, Hirai M, Nagao R, Yokota A, Takeuchi M, Yao H, Sakai K, Adachi S, Hirai H, Ashihara E, Maekawa T
  • Organizer: The American Society of Hematology 51st Annual Meeting and Exposition(ASH)
  • Place of Presentation: Ernest N. Morial Convention Center, New Orleans, USA
  • Year and Date: 2009-12-06
• [Presentation] CREB Is Involved in the Regulation of C/EBP & beta ; During'emergency' Granulopoiesis (2009)
  • Author(s): Hirai H, Kamio N, Matsusue A, Ogino S, Kimura N, Satake S, Ashihara E, Maekawa T, Tenen DG, Imanishi J
  • Organizer: The American Society of Hematology 51st Annual Meeting and Exposition
  • Place of Presentation: New Orleans, USA
  • Year and Date: 2009-12-05
• [Presentation] Overcoming CML with T315I Mutation～Detection and Therapeutics～. Kyoto University Global COE (2009)
  • Author(s): Tanaka R, Kimura S, Nagao R, Yokota A, Takeuchi M, Yao H, Sakai K, Hirai H, Ashihara E, Maekawa T
  • Organizer: Center for Frontier Medicine International Symposium/Retreat
  • Place of Presentation: Hyogo, Japan
  • Year and Date: 2009-11-07
• [Presentation] 低酸素環境とCML幹細胞厚生労働省がん研究助成金『成人白血病の難治機構の分子レベルでの解明とそれに基づく分子標的治療の開発に関する研究』班(直江班) (2009)
  • Author(s): 前川平
  • Organizer: 平成21年度第1回合同班会議
  • Place of Presentation: 名古屋
  • Year and Date: 2009-06-20
• [Patent(Industrial Property Rights)] 細胞培養容器及びその容器を用いた細胞培養方法 (2010)
  • Inventor(s): 務中達也(島津製作所)、阿部浩久(島津製作所)、叶井正樹(島津製作所)、明地将一(島津製作所)、前川平、木村晋也(佐賀大学)、芦原英司(京都府立医科大学)、庄子習一(早稲田大学)、川合健太郎(早稲田大学)
  • Industrial Property Rights Holder: 務中達也(島津製作所)、阿部浩久(島津製作所)、叶井正樹(島津製作所)、明地将一(島津製作所)、前川平、木村晋也(佐賀大学)、芦原英司(京都府立医科大学)、庄子習一(早稲田大学)、川合健太郎(早稲田大学)
  • Industrial Property Number: 特願、2010-200679
  • Filing Date: 2010-09-08