骨髄低酸素環境における骨髄腫幹細胞の解析と治療標的分子同定

• Project Area: Integrative Research on Cancer Microenvironment Network
• Project/Area Number: 23112507
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Kyoto University
• Principal Investigator: 前川 平 京都大学, 医学(系)研究科(研究院), 教授 (80229286)
• Project Period (FY): 2011-04-01 – 2013-03-31
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥8,840,000 (Direct Cost: ¥6,800,000、Indirect Cost: ¥2,040,000); Fiscal Year 2012: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2011: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
• Keywords: 多発性骨髄腫 / 低酸素 / 骨髄微小環境 / がん幹細胞 / 骨髄腫

Outline of Annual Research Achievements

多発性骨髄腫（MM）の病態維持に重要なMM幹細胞の骨髄低酸素環境の動態、および治療標的分子の同定のため、以下の検討を行った。まず正所性MM担がんマウスを作製し、MM細胞の骨髄内の動態を解析した。MM細胞は海綿骨周囲の骨内膜下に生着し、低酸素マーカーであるピモニダゾール陽性を示した。このことより、MM細胞は骨髄微小環境下では低酸素状態であることが明らかとなった。低酸素下骨髄微小環境で生存するMM細胞の性状をin vitro系で解析するため、1%低酸素状態下で長期生存可能な低酸素適応MM（HA-MM）細胞株の作製を試み、現在までに6株（AMO-1、OPM-2、NCIH929、RPMI8226、U266、IM-9）のHA-MM細胞株を樹立した。それぞれの細胞増殖を解析したところ、低酸素環境では細胞増殖スピードはそれぞれHA株で有意に遅く、またdoubling timeも有意に長いことが明らかとなった。これらの細胞株をNormoxia（20%）下の培養条件に戻したところ、増殖速度はNormoxia下での培養細胞と同等の増殖速度に復した。Ki67/7-AADを用いた二重染色による細胞周期解析では、親株であるNormoxia下での培養細胞に比して、G0期の細胞集団が有意に増加しており、さらにOct3/4、Sox2といった幹細胞マーカーのmRNAの発現亢進を認めた。CD138抗原の発現の低下は認めず、またMMの治療標的として有効であると報告してきたbeta-cateninの発現や、慢性骨髄性白血病のHA株で治療標的分子の一つとして同定したGlyoxalase 1の発現は、MM親株とHA株で発現の差は認めなかったが、TGF-betaシグナルの下流のSmad2のリン酸化亢進を認め、MMにおいてはTGF-beta/Smadシグナル系がMM幹細胞維持に関与することが示唆された。

Research Progress Status

24年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

24年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] An exploratory clinical study on the safety and efficacy of an autologous fibroblast-seeded artificial skin cultured with animal product-free medium in patients with diabetic foot ulcers (2014)
  • Author(s): Morimoto N, Ito T, Takemoto S, Katakami M, Kanda N, Tada H, Tanaka S, Teramukai S, Kawai K, Nakamura Y, Kasai Y, Yokode M, Maekawa T, Shimizu A, Suzuki S
  • Journal Title: International Wound Journal Volume: 11 Issue: 2 Pages: 183-189
  • DOI: 10.1111/j.1742-481x.2012.01064.x
  • Peer Reviewed
• [Journal Article] Combination of Ponatinib with Hedgehog Antagonist Vismodegib for Therapy-Resistant BCR-ABL1-Positive Leukemia. (2013)
  • Author(s): Katagiri S, Minami Y, Naoe T, et al.
  • Journal Title: Clin Cancer Res. Volume: 19 Issue: 6 Pages: 1422-1432
  • DOI: 10.1158/1078-0432.ccr-12-1777
  • Peer Reviewed
• [Journal Article] Effectiveness of sulforaphane as a radiosensitizer for murine osteosarcoma cells (2013)
  • Author(s): Y. Sawai, H. Murata, M. Horii, K. Koto,T. Matsui, N. Horie, Y. Tsuji, E. Ashihara, T. Maekawa, T. Kubo, S. Fushiki
  • Journal Title: Oncol Rep Volume: 29 Issue: 3 Pages: 941-945
  • DOI: 10.3892/or.2012.2195
  • Peer Reviewed
• [Journal Article] Zoledronic acid significantly enhances radiation induced apoptosis against human fibrosarcoma cells by inhibiting radioadaptive signaling. (2013)
  • Author(s): 8. Koto K, Murata H, Kimura S, Sawai Y, Horie N, Matsui T, Ryu K, Ashihara E, Maekawa T, Kubo T, Fushiki S.
  • Journal Title: Int J Oncol Volume: 42 Issue: 2 Pages: 525-534
  • DOI: 10.3892/ijo.2012.1735
  • Peer Reviewed
• [Journal Article] Activity of histone deacetylase inhibitors and an Aurora kinase inhibitor in BCR-ABL expressing leukemic cells. (2013)
  • Author(s): Okabe S
  • Journal Title: Cancer Cell International Volume: 13(1) Issue: 1 Pages: 32-40
  • DOI: 10.1186/1475-2867-13-32
  • Peer Reviewed
• [Journal Article] C/EBPβ promotes BCR-ABL-mediated myeloid expansion and leukemic stem cell exhaustion. (2013)
  • Author(s): 5. Hayashi Y, Hirai H, Kamio N, Yao H, Yoshioka S, Miura Y, Ashihara E, Fujiyama Y, Tenen DG, Maekawa T.
  • Journal Title: Leukemia Volume: 27 Issue: 3 Pages: 619-628
  • DOI: 10.1038/leu.2012.258
  • NAID: 120005466670
  • Peer Reviewed
• [Journal Article] Cyclic AMP responsive element binding proteins are involved in 'emergency' granulopoiesis through the upregulation of CCAAT/enhancer binding protein β. (2013)
  • Author(s): Hirai H
  • Journal Title: PLoS One Volume: 8 Issue: 1 Pages: e54862-e54862
  • DOI: 10.1371/journal.pone.0054862
  • NAID: 120005242481
  • Peer Reviewed
• [Journal Article] C/EBP is involved in the amplification of early granulocyte precursors during candidemia-induced ‘emergency’ granulopoiesis. (2012)
  • Author(s): Satake S
  • Journal Title: J Immunol Volume: 198 Issue: 9 Pages: 4546-4555
  • DOI: 10.4049/jimmunol.1103007
  • Peer Reviewed
• [Journal Article] Progressive graft fibrosis and donor-specific HLA antibodies in pediatric late liver allografts (2012)
  • Author(s): Miyagawa-Hayashino A, Yoshizawa A, Uchida Y, Egawa H, Yurugi K, Masuda S, Minamiguchi S, Maekawa T, Uemoto S, Haga H
  • Journal Title: Liver Transpl Volume: 18 Issue: 11 Pages: 1333-1342
  • DOI: 10.1002/lt.23534
  • Peer Reviewed
• [Journal Article] Effects of low-intensity pulsed ultrasound (LIPUS) on osteosarcoma and cancer cells. (2012)
  • Author(s): Sawai Y
  • Journal Title: Oncol Rep Volume: 28 Issue: 2 Pages: 481-486
  • DOI: 10.3892/or.2012.1816
  • Peer Reviewed
• [Journal Article] Incidence and pattern of hemolytic anemia after minor ABO-mismatched living- d lobar lung transplantation. (2012)
  • Author(s): Ohsumi A
  • Journal Title: Surg Today Volume: 43 Issue: 11 Pages: 1250-1253
  • DOI: 10.1007/s00595-012-0422-3
  • Peer Reviewed
• [Journal Article] III. 造血器腫瘍の診断と治療 治療法 支持療法；輸血副作用とHLA抗体. (2012)
  • Author(s): 吉岡聡
  • Journal Title: 日本臨床 Volume: 70 Pages: 211-245
• [Journal Article] 細胞周期調節系 (2012)
  • Author(s): 前川 平
  • Journal Title: 日本臨牀 Volume: 70(suppl) Pages: 119-124
• [Journal Article] Phase I/II a clinical trials of dendritic cell-based immunotherapy for acute myeloid leukemia in elderly patients using autologous apoptotic leukemic cells or a heteroclitic WT1 peptide as antigens (2011)
  • Author(s): Kitawaki T, Kadowaki, T, Maekawa T, et al
  • Journal Title: Brit J Haematol Volume: 153 Issue: 6 Pages: 796-799
  • DOI: 10.1111/j.1365-2141.2010.08490.x
  • Peer Reviewed
• [Journal Article] AV-65, a novel Wnt/β-catenin signal inhibitor, successfully suppresses progression of multiple myeloma in a mouse model (2011)
  • Author(s): Yao H, Ashihara E, Maekawa T, et al
  • Journal Title: Blood Cancer Journal Volume: 1 Issue: 11 Pages: e43-e43
  • DOI: 10.1038/bcj.2011.41
  • Peer Reviewed
• [Journal Article] Galectin-3(Gal-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, Shimura Y, Kiyota M, Nakayama R, Akaji K, Taki T, Uoshima N, Kobayashi Y, Horiike S, Maekawa T, Taniwaki M
  • Journal Title: Proc Natl Acad Sci U S A Volume: 108 Issue: 42 Pages: 17468-17473
  • DOI: 10.1073/pnas.1111138108
  • Peer Reviewed
• [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) Issue: 2 Pages: 228-34
  • DOI: 10.1016/j.canlet.2011.08.009
  • Peer Reviewed
• [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) Issue: 1 Pages: 91-100
  • DOI: 10.1016/j.canlet.2011.08.002
  • Peer Reviewed
• [Journal Article] Rakicidin A effectively induces apoptosis in hypoxia adapted Bcr-Abl positive leukemic cells (2011)
  • Author(s): Takeuchi M, Ashihara E, Maekawa T, et al
  • Journal Title: Cancer Sci Volume: 102 Issue: 3 Pages: 591-6
  • DOI: 10.1111/j.1349-7006.2010.01813.x
  • NAID: 10029291246
  • Peer Reviewed
• [Journal Article] Galection-9 ameliorates acute Graft-versus-host disease through the induction of T-cell apoptosis (2011)
  • Author(s): Sakai K, Ashihara E, Maekawa T, et al
  • Journal Title: Eur J Immunol Volume: 41 Issue: 1 Pages: 67-75
  • DOI: 10.1002/eji.200939931
  • Peer Reviewed
• [Journal Article] Cross-priming of CD8(+) T cells in vivo by dendritic cells pulsed with autologous apoptotic leukemic cells in immunotherapy for elderly patients with acute myeloid leukemia. (2011)
  • Author(s): Kitawaki, T., Kadowaki, N., Fukunaga, K., Kasai, Y., Maekawa, T., Ohmori, T., Itoh, T., Shimizu, A., Kuzushima, K., Kondo, T., Ishikawa, T.and Uchiyama, T.
  • Journal Title: Exp. Hematol. Volume: 39 Issue: 4 Pages: 424-433
  • DOI: 10.1016/j.exphem.2011.01.001
  • NAID: 120002970837
  • Peer Reviewed
• [Journal Article] Targeting the Wnt/β-catenin signaling pathway in human cancers (2011)
  • Author(s): Yao H, Ashihara E, Maekawa T
  • Journal Title: Expert Opinion on Therapeutic Targets Volume: 15 Issue: 7 Pages: 873-887
  • DOI: 10.1517/14728222.2011.577418
  • Peer Reviewed
• [Journal Article] RNA interference against Polo-like kinase-1 in advanced non-small cell lung cancers (2011)
  • Author(s): Kawata E, Ashihara E, Maekawa T
  • Journal Title: J Clin Bioinformatics Volume: 1 Issue: 1 Pages: 6-6
  • DOI: 10.1186/2043-9113-1-6
  • Peer Reviewed
• [Presentation] C/EBPβ is upregulated through STAT5 and accelerates exhaustion of leukemic stem cells in chronic phase of chronic myeloid leukemia. (2012)
  • Author(s): Hayashi Y
  • Organizer: The 54th ASH Annual Meeting and Exposition
  • Place of Presentation: Atlanta, GA, USA
  • Year and Date: 2012-12-11
• [Presentation] Expression of C/EBPβ in Bone Marrow Mesenchymal Stem Cells Is Mandatory for Early-Stage B Cell Lymphopoiesis. (2012)
  • Author(s): Yoshioka S
  • Organizer: The 54th ASH Annual Meeting and Exposition
  • Place of Presentation: Atlanta, GA, USA
  • Year and Date: 2012-12-10
• [Presentation] Mesenchymal stem cells skewed their phenotype toward the osteogenic lineage support the hematopoietic cell differentiation. (2012)
  • Author(s): Yao H
  • Organizer: The 54th ASH Annual Meeting and Exposition
  • Place of Presentation: Atlanta, GA, USA
  • Year and Date: 2012-12-10
• [Presentation] C/EBPβ-mediated expansion of hematopoietic stem/progenitors precedes ‘emergency’ granulopoiesis induced by candidemia (2012)
  • Author(s): Tamura A
  • Organizer: The 54th ASH Annual Meeting and Exposition
  • Place of Presentation: Atlanta, GA, USA
  • Year and Date: 2012-12-09
• [Presentation] Myeloid-derived suppressor cells accumulated in soleens of mice with collagen-induced arthritis and inhibited immune response od CD4+ T cells. (2012)
  • Author(s): Fujii W
  • Organizer: American College of Rheumatology 2012 Annual Scientific Meeting
  • Place of Presentation: Washington, DC, USA
  • Year and Date: 2012-11-13
• [Presentation] Defective mesenchymal stem cells contribute to impaired B cell lymphopoiesis in C/EBPb knockout mice. (2012)
  • Author(s): Yoshioka S
  • Organizer: The 74th Annual Meeting of the Japanese Society of Hematology
  • Place of Presentation: Kyoto, Japan
  • Year and Date: 2012-10-20
• [Presentation] Co-treatment with ponatinib and HDAC inhibitor vorinostat: A treatment for Ph-positive cells. (2012)
  • Author(s): Okabe S
  • Organizer: The 74th Annual Meeting of the Japanese Society of Hematology
  • Place of Presentation: Kyoto, Japan
  • Year and Date: 2012-10-20
• [Presentation] Targeting the Hedgehog pathway in therapy-resistant BCR-ABL1 positive leukemia with ponatinib. (2012)
  • Author(s): Katagiri S
  • Organizer: The 74th Annual Meeting of the Japanese Society of Hematology
  • Place of Presentation: Kyoto, Japan
  • Year and Date: 2012-10-20
• [Presentation] Combined effects of ponatinib and dasatinib against T315I mutant forms of BCR-ABL1. (2012)
  • Author(s): Tauchi T
  • Organizer: The 74th Annual Meeting of the Japanese Society of Hematology
  • Place of Presentation: Kyoto, Japan
  • Year and Date: 2012-10-20
• [Presentation] Involvement of C/EBPβ in the pathogenesis of chronic phase chronic myeloid leukemia. (2012)
  • Author(s): Hayashi Y
  • Organizer: The 74th Annual Meeting of the Japanese Society of Hematology
  • Place of Presentation: Kyoto, Japan
  • Year and Date: 2012-10-20
• [Presentation] Parathyroid hormone stimulates mesenchymal stem cells to enhance expansion of hematopoietic cells. (2012)
  • Author(s): Yao H
  • Organizer: The 74th Annual Meeting of the Japanese Society of Hematology
  • Place of Presentation: Kyoto, Japan
  • Year and Date: 2012-10-19
• [Presentation] BCR-ABL による C/EBPβの発現亢進が慢性骨髄性白血病の骨髄球系造血亢進および病態進行をもたらす. (2012)
  • Author(s): 林 嘉宏
  • Organizer: 第71回日本癌学会学術総会
  • Place of Presentation: 札幌
  • Year and Date: 2012-09-20
• [Presentation] 新規Wnt/beta-カテニン経路阻害剤AV65による多発性骨髄腫細胞の増殖抑制効果. (2012)
  • Author(s): 芦原英司
  • Organizer: 第71回日本癌学会学術総会
  • Place of Presentation: 札幌
  • Year and Date: 2012-09-19
• [Presentation] 原発性胆汁性肝硬変患者が保有するＨＬＡ抗体の特異性について. (2012)
  • Author(s): 万木紀美子
  • Organizer: 第21回日本組織適合性学会大会
  • Place of Presentation: 東京都
  • Year and Date: 2012-09-16
• [Presentation] An exploratory clinical study on the safety and efficacy of an autologous fibroblast-seeded artificial skin cultured with animal-product free medium in patients with diabetic foot ulcers. (2012)
  • Author(s): Morimoto N
  • Organizer: 4th Congress of the World Union of Wound Healing Societies
  • Place of Presentation: Yokohama, Japan
  • Year and Date: 2012-09-02
• [Presentation] Successful co-treatment of BCR-ABL leukemic cells with ponatinib, an ABL kinase inhibitor, and vorinostat, a histone deacetylase inhibitor: A potential treatment for BCR-ABL positive leukemia cells. (2012)
  • Author(s): Okabe S
  • Organizer: 17th Congress of European Society of Hematology
  • Place of Presentation: Amsterdam, The Netherland
  • Year and Date: 2012-07-14
• [Presentation] BCR/ABLの白血病原性における転写因子 C/EBPβの関与. (2012)
  • Author(s): 林 嘉宏
  • Organizer: 第16回日本がん分子標的治療学会学術集会
  • Place of Presentation: 福岡
  • Year and Date: 2012-06-28
• [Presentation] BCR/ABL-mediated myeloid expresion is promoted by C/EBPβ, a regulator of emergency granulopoiesis (2011)
  • Author(s): Hayashi Y, Hirai H, Maekawa T, et al
  • Organizer: 第53回アメリカ血液学会
  • Place of Presentation: 米国、サンディエゴ
  • Year and Date: 2011-12-12
• [Presentation] Parathyroid hormone stimulates human mesenchymal stem cells to enhance the expansion of hematopoietic stem cells (2011)
  • Author(s): Yao H, Miura Y, Maekawa T, et al
  • Organizer: 第53回アメリカ血液学会
  • Place of Presentation: 米国、サンディエゴ
  • Year and Date: 2011-12-11
• [Presentation] Combined effects of a pan-Abl1 kinase inhibitor, ponatinib and dasatinib against T315I mutant forms of Bcr-Abl1 : in vitro and in vivo studies (2011)
  • Author(s): Tauchi T, Maekawa T, Ohyashiki K, et al
  • Organizer: 第53回アメリカ血液学会
  • Place of Presentation: 米国、サンディエゴ
  • Year and Date: 2011-12-11
• [Presentation] Targeting the hedgehog signaling pathway in therapy-resistant Bcr-Abl1 positive leukemia with ponatinib (2011)
  • Author(s): Katagiri S, Maekawa T, Ohyashiki K, et al
  • Organizer: 第53回アメリカ血液学会
  • Place of Presentation: 米国、サンディエゴ
  • Year and Date: 2011-12-11
• [Presentation] Combined effects of the hedgehog inhibitor LDE225 and nilotinib in a random mutagesesis screen (2011)
  • Author(s): Tauchi T, Okabe S, Maekawa T, et al
  • Organizer: 第73回日本血液学会学術集会
  • Place of Presentation: 名古屋国際会議場
  • Year and Date: 2011-10-15
• [Presentation] Involvement of C/EBPβ in BCR/ABL-dependent leukemogenesis (2011)
  • Author(s): Hayashi Y, Hirai H, Maekawa T, et al
  • Organizer: 第73回日本血液学会学術集会
  • Place of Presentation: 名古屋国際会議場
  • Year and Date: 2011-10-15
• [Presentation] Cord blood transplantation for T315I-mutated blastic-phase chronic myeloid leukemia (2011)
  • Author(s): Kunitomi A, Kotani S, Maekawa T, et al
  • Organizer: 第73回日本血液学会学術集会
  • Place of Presentation: 名古屋国際会議場
  • Year and Date: 2011-10-15
• [Presentation] A new BCR/ABL-positive megakaryoblastic cell line resistant to both imatinib and dasatinib (2011)
  • Author(s): Shimazu Y, Hishizawa M, Maekawa T, et al
  • Organizer: 第73回日本血液学会学術集会
  • Place of Presentation: 名古屋国際会議場
  • Year and Date: 2011-10-14
• [Presentation] Galectin-3 induced by leukemia microenvironment promotes drug resistance and BM lodgment in CML (2011)
  • Author(s): Yamamoto M, Kuroda J, Maekawa T, et al
  • Organizer: 第73回日本血液学会学術集会
  • Place of Presentation: 名古屋国際会議場
  • Year and Date: 2011-10-14
• [Presentation] AV-65 inhibited the growth of myeloma cells by enhancing β-TrCP-mediated β-catenin ubiquitination (2011)
  • Author(s): Yao H, Ashihara E, Maekawa T, et al
  • Organizer: 第73回日本血液学会学術集会
  • Place of Presentation: 名古屋国際会議場
  • Year and Date: 2011-10-14
• [Presentation] A microfluidic device to mimic the bone marrow environment : real-time observation of the leukemic cell behavior (2011)
  • Author(s): Munaka T, Ashihara E, Maekawa T, et al
  • Organizer: The 15^<th> Intemational Conference on Miniaturized Systems for Chemistry and Life Sciences
  • Place of Presentation: 米国、シアトル
• [Book] みんなに役立つ造血幹細胞移植の基礎と臨床#8212;改訂版 (2012)
  • Author(s): 前川 平
  • Total Pages: 8
  • Publisher: 医薬ジャーナル社
• [Remarks] 京都大学医学部附属病院輸血細胞治療部
  • URL: http://dtm.kuhp.kyoto-u.ac.jp/
• [Remarks]
  • URL: http://dtm.duhp.kyoto-u.ac.jp/
• [Remarks]
  • URL: http://www.kuhp.kyoto-u.ac.jp/~ccmt/