骨髄低酸素環境における骨髄腫幹細胞維持分子を標的とした新規治療開発

• Project Area: Integrative Research on Cancer Microenvironment Network
• Project/Area Number: 25112706
• 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): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥7,540,000 (Direct Cost: ¥5,800,000、Indirect Cost: ¥1,740,000); Fiscal Year 2014: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000); Fiscal Year 2013: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
• Keywords: 多発性骨髄腫 / 骨髄腫幹細胞 / 低酸素環境 / Wnt/β-catenin経路 / 骨髄微小環境 / がん幹細胞

Outline of Annual Research Achievements

【研究経緯と目的】多発性骨髄腫 (MM) に対する治療成績は、近年の造血幹細胞移植術の進歩やボルテゾミブやレナリドマイドなどの新規治療薬の開発により急速に向上してきた。しかし根治できるMMはごく一部の症例に限られ、その原因の一つとして、MM細胞を供給する幹細胞が残存することが指摘されている。本研究では、①低酸素環境下にあるMM幹細胞の性状・動態を解析し、②MM根治に向けた標的分子を探索することを目的とした。
【研究実績の概要】①低酸素環境適応MM（MM-HA）幹細胞の性状解析：MM細胞株を低酸素(1%02)環境下で長期間培養して、AMO-1/HA、OPM-2/HA、IM-9/HAの３つの低酸素適応細胞株を樹立した(MM-HA細胞株)。MM-HA幹細胞の性状解析では、SP分画やGo期の細胞が多く、β-cateninの発現が高いなど、いわゆる”幹細胞性”を示すことを確認した。②細胞増殖はHA株で有意に遅く、Ki67/7-AADによる細胞周期解では、7-AADの蛍光強度が2nに存在するG0/G1分画のうち、Ki67陰性のG0期の細胞集団は、HA株において親株より有意に多く存在した。これらの細胞株のうち、AMO-1親株とHA株を用いて正所性MM担癌モデルマウスを作製し生存期間の検討した。HA株を移植したマウスにおいて有意に生存期間の短縮を認めた。これらのことからMM-HA株も、いわゆる“がん幹細胞”の性質を有することが示唆された。次に、我々がMMの新たな治療標的として明らかにしてきたWnt/β-catenin経路について検討した。HA株においてβ-cateninの発現増強やWnt/β-catenin/TCFシグナルの亢進は認めず、HA株のMM幹細胞性にはWnt経路は関与していないことが推察された。今後、MM幹細胞維持に重要な経路について網羅的に解析し標的分子を同定したい。

Research Progress Status

26年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

26年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Targeting the canonical Wnt /β-catenin pathway in hematological malignancies. (2015)
  • Author(s): Ashihara E, Takada T, Maekawa T
  • Journal Title: Cancer Sci Volume: - Issue: 6 Pages: 665-671
  • DOI: 10.1111/cas.12655
  • NAID: 120005767015
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] CCR1-mediated accumulation of myeloid cells in the liver microenvironment promoting mouse colon cancer metastasis (2014)
  • Author(s): Hirai H, Fujishita T, Kurimoto K, Miyachi H, Kitano S, Inamoto S, Itatani Y, Saitou M, Maekawa T, Taketo MM
  • Journal Title: Clin Exp Metastasis Volume: 31 Issue: 8 Pages: 977-989
  • DOI: 10.1007/s10585-014-9684-z
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] CCR1-mediated Accumulation of Myeloid Cells in the Liver Microenvironment Promoting Mouse Colon Cancer Metastasis (2014)
  • Author(s): Hirai H, et al
  • Journal Title: Clinical and Experimental Metastasis Volume: 32 Issue: 8 Pages: 2245-55
  • DOI: 10.1002/stem.1701
  • Peer Reviewed / Open Access
• [Journal Article] CCAAT/Enhancer-Binding Protein β Expressed by Bone Marrow Mesenchymal Stromal Cells Regulates Early B-Cell Lymphopoiesis (2014)
  • Author(s): Yoshioka S, Miura Y, Yao H, Satake S, Hayashi Y, Tamura A, Hishita T, Ichinohe T, Hirai H, Takaori-Kondo A, Maekawa T
  • Journal Title: Stem Cells Volume: 32 Issue: 3 Pages: 730-740
  • DOI: 10.1002/stem.1555
  • Peer Reviewed
• [Journal Article] Efficacy of the dual PI3K and mTOR inhibitor NVP-BEZ in combination with nilotinib against BCR-ABL-positive leukemia cells involves the ABL kinase domain mutation. (2014)
  • Author(s): Okabe S, Tauchi T, Tanaka Y, Kitahara T, Kimura S, Maekawa T, Ohyashiki K
  • Journal Title: Cancer Biol Ther Volume: 15 Issue: 2 Pages: 207-215
  • DOI: 10.4161/cbt.26725
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Two Susceptibility Loci to Takayasu Arteritis Reveal a Synergistic Role of the IL12B and HLA-B Regions in a Japanese Population (2013)
  • Author(s): Terao C, Yoshifuji H, Kimura A, Matsumura T, Ohmura K, Takahashi M, Shimizu M, Kawaguchi T, Chen Z, Naruse TK, Sato-Otsubo A, Ebana Y, Maejima Y, Kinoshita H, Murakami K, Kawabata D, Wada Y, Narita I,
  • Journal Title: American Journal of Human Genetics Volume: 93 Issue: 2 Pages: 289-297
  • DOI: 10.1016/j.ajhg.2013.05.024
  • 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] Chimerism of bone marrow mesenchymal stem/stromal cells in allogeneic hematopoietic cell transplantation: is it clinically relevant? (2013)
  • Author(s): Miura Y, Yoshioka S, Yao H, Takaori-Kondo A, Maekawa T, Ichinohe T.
  • Journal Title: Chimerism Volume: 4(3) Issue: 3 Pages: 78-83
  • DOI: 10.4161/chim.25609
  • 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] 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] Loss of SMAD4 from colorectal cancer cells promotes CCL15 expression to recruit CCR1+ myeloid cells and facilitate liver metastasis. (2013)
  • Author(s): Itatani Y, Kawada K, Fujishita T, Kakizaki F, Hirai H, Matsumoto T, Iwamoto M, Inamoto S, Hatano E, Hasegawa S, Maekawa T, Uemoto S, Sakai Y, Taketo MM
  • Journal Title: Gastroenterol Volume: 145(5) Issue: 5 Pages: 1064-1075
  • DOI: 10.1053/j.gastro.2013.07.033
  • NAID: 120005348333
  • Peer Reviewed
• [Journal Article] Myeloid-Derived Suppressor Cells Play Crucial Roles in the Regulation of Mouse Collagen-Induced Arthritis. (2013)
  • Author(s): 1. Fujii W, Ashihara E, Hirai H, Nagahara H, Kajitani N, Fujioka K, Murakami K, Seno T, Yamamoto A, Ishino H, Kohno M, Maekawa T, Kawahito Y.
  • Journal Title: J Immunol Volume: 191 Issue: 3 Pages: 1073-1081
  • DOI: 10.4049/jimmunol.1203535
  • 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, Kamio N, Huang G, Matsusue A, Ogino S, Kimura N, Satake S, Ashihara E, Imanishi J, Tenen DG ,Maekawa T
  • Journal Title: PLoS ONE Volume: 8
  • 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] Association of Takayasu arteritis with HLA-B*67:01 and two amino acids in HLA-B protein. (2013)
  • Author(s): Terao C, Yoshifuji H, Ohmura K, Murakami K, Kawabata D, Yurugi K, Tazaki J, Kinoshita H, Kimura A, Akizuki M, Kawaguchi Y, Yamanaka H, Miura Y, Maekawa T, Saji H, Mimori T
  • Journal Title: Rheumatology Volume: 52(10) Issue: 10 Pages: 1769-1774
  • DOI: 10.1093/rheumatology/ket241
  • 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
• [Presentation] CMLに対するIFNαの作用における転写因子C/EBPβの関与. (2015)
  • Author(s): 横田明日美、平位秀世、林嘉宏、田村彰広、佐藤淳至、岩佐麿佐紀、藤城綾、庄司月美、三浦康生、前川 平
  • Organizer: 平成26年度文部科学省新学術領域研究 がん研究分野の特性等を踏まえた支援活動 公開シンポジウム
  • Place of Presentation: 東京
  • Year and Date: 2015-01-28
• [Presentation] ABLファミリー遺伝子間での生理作用の違いから検討できる分子機構とその意義. (2015)
  • Author(s): 奥田恵子、横田明日美、庄司月美、平位秀世、前川平
  • Organizer: 第19回造血器腫瘍研究会
  • Place of Presentation: 佐賀
  • Year and Date: 2015-01-24
• [Presentation] C/EBPbetaによる造血幹細胞制御. (2015)
  • Author(s): 平位秀世、佐藤淳至、田村彰広、横田明日美、庄司月美、三浦康生、志馬伸朗、前川 平
  • Organizer: 第19回造血器腫瘍研究会
  • Place of Presentation: 佐賀
  • Year and Date: 2015-01-23
• [Presentation] CCR1-mediated formation of liver microenvironment for colon cancer metastasis (2015)
  • Author(s): Hirai H, Fujishita T, Kurimoto K, Saitou M, Maekawa T,Taketo MM
  • Organizer: 日本癌学会シンポジウム/共同利用・共同研究拠点シンポジウム「がん幹細胞・微小環境・分子標的～がん進展制御への挑戦」
  • Place of Presentation: 金沢
  • Year and Date: 2015-01-21
• [Presentation] Hypoxia-adapted myeloma cells possess stem cell characters. (2015)
  • Author(s): Ashihara E, Nakagawa Y, Yao H, Takada T, Yokota A, Miura Y, Takata K, Hirai H, Kitamura Y, Maekawa T
  • Organizer: 日本癌学会シンポジウム/共同利用・共同研究拠点シンポジウム「がん幹細胞・微小環境・分子標的～がん進展制御への挑戦」
  • Place of Presentation: 金沢
  • Year and Date: 2015-01-21
• [Presentation] 低酸素環境に潜む白血病・骨髄腫幹細胞の殲滅を目指した革新的治療法の開発 (2015)
  • Author(s): 前川 平
  • Organizer: がん微小環境ネットワークの統合的研究（文部科学省・科学研究費補助金・新学術領域研究）班会議
  • Place of Presentation: つくば国際会議場
  • Year and Date: 2015-01-14
• [Presentation] Essential Roles of C/EBPβ in Survival of Ly6C- monocytes. (2014)
  • Author(s): Tamura A, Hirai H, Yokota A, Sato A, Yao H, Iwasa M, Fujishiro A, Miura Y, Maekawa T
  • Organizer: American Society of Hematology, 56th Annual Meeting
  • Place of Presentation: San Francisco, CA, USA
  • Year and Date: 2014-12-08
• [Presentation] CCR1-Mediated Formation of Liver Microenvironment for Cancer Metastasis. (2014)
  • Author(s): Hirai H, Fujishita T, Kurimoto K, Saitou M, Maekawa T, Mark M Taketo
  • Organizer: American Society of Hematology, 56th Annual Meeting
  • Place of Presentation: San Francisco, CA, USA
  • Year and Date: 2014-12-08
• [Presentation] C-Terminal Domain of ABL Family Kinases, ABL and ARG, Defines Their Distinct Leukemogenic Activities in Vivo. (2014)
  • Author(s): Yokota A, Hirai H, Shoji T, Maekawa T, Okuda K
  • Organizer: American Society of Hematology, 56th Annual Meeting
  • Place of Presentation: San Francisco, CA, USA
  • Year and Date: 2014-12-07
• [Presentation] Effect of irradiation on the characteristics of human bone marrow mesenchymal stromal/stem cells. (2014)
  • Author(s): Iwasa M, Miura Y, Yao H, Yoshioka S, Fujishiro A, Tamura A, Sato A, Yokota A, Hirai H, Ando A, Ichinohe T, Maekawa T
  • Organizer: The 76th Annual Meeting of the Japanese Society of Hematology （第７６回日本血液学会学術総会)
  • Place of Presentation: 大阪
  • Year and Date: 2014-11-01
• [Presentation] Panovinostat and Ponatinib show synergistic cytotoxicity against Imatinib-resistant CML cell lines. (2014)
  • Author(s): Matsuda Y, Yamauchi T, Shigemi H, Negoro E, Morinaga K, Uzui K, Nishi R, Kimua S, Maekawa T, Ueda T
  • Organizer: The 76th Annual Meeting of the Japanese Society of Hematology （第７６回日本血液学会学術総会）
  • Place of Presentation: 大阪
  • Year and Date: 2014-11-01
• [Presentation] solation of mesenchymal stromal/stem cells from UCB ineligible for public UCB donation. (2014)
  • Author(s): Yoshioka S, Miura Y, Yao H, Iwasa M, Fujishiro A, Higashi Y, Yokota A, Tamura A, Sato A, Hirai H, Ichinohe T, Takaori-Kondo A, Maekawa T
  • Organizer: The 76th Annual Meeting of the Japanese Society of Hematology （第７６回日本血液学会学術総会)
  • Place of Presentation: 大阪
  • Year and Date: 2014-11-01
• [Presentation] C/EBPβpromotes initial expansion and exhaustion of hematopoietic stem cells in response to stresses. (2014)
  • Author(s): Sato A, Hirai H, Tamura A, Yokota A, Hayashi Y, Yao H, Yoshiioka S, Iwasa M, Miura Y, Maekawa T
  • Organizer: The 76th Annual Meeting of the Japanese Society of Hematology (第７６回日本血液学会学術総会)
  • Place of Presentation: 大阪
  • Year and Date: 2014-10-31
• [Presentation] Critical roles of C/EBP β for survival of Ly6C-Monocytes. (2014)
  • Author(s): Tamura A, Hirai H, Yokota A, Sato A, Yao H, Yoshioka S, Iwasa M, Miura Y, Ashi
  • Organizer: The 76th Annual Meeting of the Japanese Society of Hematology （第７６回日本血液学会学術総会)
  • Place of Presentation: 大阪
  • Year and Date: 2014-10-31
• [Presentation] : Involvement of C/EBP β in IFN α-induced differentiation of BCR-ABL-expressing leukemic cells. (2014)
  • Author(s): Yokota A, Hirai H, Hayashi Y, Tamura A, Sato A, Yao H,Yoshioka S, Iwasa M, Miura Y, Maekawa T
  • Organizer: The 76th Annual Meeting of the Japanese Society of Hematology （第７６回日本血液学会学術総会)
  • Place of Presentation: 大阪
  • Year and Date: 2014-10-31
• [Presentation] BCR-ABL発現白血病細胞に対するC/EBPβを介したIFNαの作用. (2014)
  • Author(s): 横田明日美、平位秀世、林 嘉宏、田村彰広、佐藤淳至、八尾尚幸、岩佐麿佐紀、藤城 綾、三浦康生、前川 平
  • Organizer: 第73回日本癌学会学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2014-09-26
• [Presentation] Parathyroid hormone enhances expansion through CDH11 in human bone marrow mesenchymal stromal/stem cells. (2013)
  • Author(s): Yao H, Miura Y, Yoshioka S, Hayashi Y, Yokota A, Tamura A, Ichinohe T, Hirai H and Maekawa T.
  • Organizer: AsiaCORD 2013, Scientific Symposium I-1 "New Insights into Stem Cell Biology- Stem Cell Development & Differentiation"
  • Place of Presentation: Kobe, Japan
• [Presentation] C/EBPβ in mesenchymal stromal/stem cells regulates early B cell lymphopoiesis. (2013)
  • Author(s): Yoshioka S, Miura Y, Yao H, Hayashi Y, Tamura A, Hishita T, Ichinohe T, Hirai H, Takaori-Kondo A and Maekawa T.
  • Organizer: AsiaCORD 2013, Scientific Symposium I-1 "New Insights into Stem Cell Biology- Stem Cell Development & Differentiation"
  • Place of Presentation: Kobe, Japan
  • Invited
• [Presentation] Myeloid-derived suppressor cells have regulatory roles in mouse collagen-induced arthritis. (2013)
  • Author(s): Fujii W, Ashihara E, Hirai H, Nagahara H, Kajitani N, Fujioka K, Murakami K, Seno T, Yamamoto A, Ishino H, Kohno M, Maekawa T, Kawahito Y.
  • Organizer: Annual European Congress of Rheumatology 2013.
  • Place of Presentation: Madrid, Spain
  • Invited
• [Presentation] Impact of donor-specific anti-HLA and anti-angiotensin-II type 1 receptor antibodies on graft fibrosis after immunosuppression withdrawal in pediatric liver transplantation. (2013)
  • Author(s): Ohe H, Uchida Y, Yoshizawa A, Hirao H, Taniguchi M, Maruya E, Maekawa T, Uemoto S, Terasaki PI.
  • Organizer: The 13th Congress of the Asian Society of Transplantation
  • Place of Presentation: Kyoto, Japan
  • Invited
• [Presentation] C/EBPβ expressed by bone marrow mesenchymal stromal cells is indispensable for precursor B-cell lymphopoiesis. (2013)
  • Author(s): Yoshioka S, Miura Y, Yao H, Iwasa M, Sato A, Tamura A, Yokota A, Hishita T, Ichinohe T, Hirai H, Maekawa T.
  • Organizer: 55th American Society of Hematology
  • Place of Presentation: New Orleans, LA, USA
  • Invited
• [Presentation] Cytokine-STATs signalings upregulate endogenous C/EBPβ in BCR-ABL+ leukemic cells independently from BCR-ABL signaling. (2013)
  • Author(s): Yokota A, Hirai H, Hayashi Y, Tamura A, Sato A, Yao H, Yoshioka S, Iwasa M, Ashihara E, Miura Y, Maekawa T.
  • Organizer: 55th American Society of Hematology
  • Place of Presentation: New Orleans, LA, USA
  • Invited
• [Presentation] Cell-intrinsic and -extrinsic involvement of C/EBPβ in the regulation of hematopoietic stem cells. (2013)
  • Author(s): Tamura A, Hirai H, Hayashi Y, Yokota A, Sato A, Yao H, Yoshioka S, Iwasa M, Ashihara E, Miura Y, Maekawa T.
  • Organizer: 55th American Society of Hematology
  • Place of Presentation: New Orleans, LA, USA
  • Invited
• [Presentation] Hypoxia-Adapted Myeloma Cells Possess Stem Cell Character. (2013)
  • Author(s): Ashihara E, Nakagawa Y, Yao H, Yokota A, Miura Y, Takata K, Kitamura Y, Hirai H , Maekawa T.
  • Organizer: 55th American Society of Hematology
  • Place of Presentation: New Orleans, LA, USA
  • Invited
• [Book] Emerging Trends in Cell and Gene Therapy. (2013)
  • Author(s): Ashihara E, Maekawa T
  • Total Pages: 705
  • Publisher: Springe, New York Heidelberg Dordrecht London
• [Remarks] 京都大学医学部附属病院輸血細胞治療部 研究年報・業績 2013年
  • URL: http://dtm.kuhp.kyoto-u.ac.jp/report