A unique anti-leukemic therapy using efficiently tRNA precursor cleavage enzyme in leukemia cells

• Project/Area Number: 24591385
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Hematology
• Research Institution: Niigata University
• Principal Investigator: Takahashi Masuhiro 新潟大学, 医歯学系, 教授 (90179531)
• Co-Investigator(Kenkyū-buntansha): 鳥羽 健 新潟大学, 医歯学総合病院, 講師 (60313540)
• Project Period (FY): 2012-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2013: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2012: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
• Keywords: TRUE gene silencing / tRNase ZL / Bcl-2 / small guide RNA / 白血病 / アポトーシス誘導 / 細胞増殖抑制 / マウス移植実験 / sgRNA / 腫瘍抗原タンパク / アポソーシス誘導 / 核酸医薬 / tRNaseZL / 腫瘍抗原タンパ / がん抗原蛋白 / アポトーシス

Outline of Final Research Achievements

TRUE gene silencing is a technology to eliminate specific cellular RNAs by using tRNase ZL and small guide RNA (sgRNA). Here we investigated how WT1-mRNA-targeting sgRNAs affect leukemic cells. We showed that sgRNA can be easily taken up by cells without any transfection reagents, and that the naked sgRNAs targeting the WT1 mRNA can reduce its mRNA levels and WT1 protein amounts in the WT1-expressing leukemic cells. Concomitantly, these sgRNAs efficiently induced apoptosis in these cells but not in WT1-nonexpressing cells. We also demonstrated that the reduction in the WT1 mRNA level is caused by its cleavage by tRNase ZL.
Besides, we demonstrated that a heptamer, mh1(Bcl-2), which targets the human Bcl-2 mRNA, can be taken up by cells without any transfection reagents and that it can induce apoptosis of the leukemia cells. Mouse xenograft experiments showed that a median survival of the mh1(Bcl-2)-treated mice was longer than that of the control mice.

Research Products

• [Journal Article] Five years molecular remission with an increase of WT1-specific CTLs after cessation of WT1 peptide vaccination in a patient with CML. (2016)
  • Author(s): Narita M, Masuko M, Takahashi M
  • Journal Title: Int J Med Sci Volume: 13 Issue: 4 Pages: 0-0
  • DOI: 10.7150/ijms.13601
  • Peer Reviewed / Open Access
• [Journal Article] Immune responses in patients with esophageal cancer treated with SART1 peptide-pulsed dendritic cell vaccine. (2015)
  • Author(s): Narita M, Kanda T, Abe T, Uchiyama T, Iwafuchi M, Zheng Z, Liu A, Kaifu T, Kosugi S, Minagawa M, Itoh K, Takahashi M.
  • Journal Title: Int J Oncol. Volume: 46 Issue: 4 Pages: 1699-1709
  • DOI: 10.3892/ijo.2015.2846
  • Peer Reviewed / Open Access
• [Journal Article] Screening of a heptamer-type sgRNA library for potential therapeutic agents against hematological malignancies. (2014)
  • Author(s): Takahashi M, Elbarbary RA, Watanabe N, Goto A, Kamiya D, Watabe Y, Uchiyama T, Narita M, Takahashi M, Takahashi Y, Ishihara N, Miyazawa T, Yoshida T, Kawano M, Tamura M, Nashimoto M.
  • Journal Title: Leuk Res Volume: 38 Issue: 2 Pages: 808-815
  • DOI: 10.3892/mmr.2015.3685
  • Peer Reviewed / Open Access
• [Journal Article] Ustekinumab improves psoriasis without suppressing tumor antigen-specific cytotoxic T lymphocytes. (2014)
  • Author(s): Narita M, Nishizawa Y, Iwaya S, Oiwa E, Iwabuchi M, Uchiyama T, Matsuyama A, Masuko M, Takahashi M
  • Journal Title: Int Arch Allergy Immunol. Volume: 165 Issue: 1 Pages: 52-60
  • DOI: 10.1159/000366503
  • Peer Reviewed / Open Access
• [Journal Article] Activation of the leukemia plasmacytoid dendritic cell line PMDC05 by Toho-1, a novel IDO inhibitor. (2014)
  • Author(s): Yamahira A, Narita M, Iwabuchi M, Uchiyama T, Iwaya S, Ohiwa R, Nishizawa Y, Suzuki T, Yokoyama Y, Hashimoto S, Takizawa J, Sone H, Takahashi M.
  • Journal Title: Anticancer Res. Volume: 34 Pages: 4021-4028
  • Peer Reviewed / Open Access
• [Journal Article] Induction of apoptosis of leukemic cells by TRUE gene silencing using small guide RNAs targeting the WT1 mRNA. (2013)
  • Author(s): Watanabe N, Narita M, Yamahira A, Taniguchi T, Furukawa T, Yoshida T, Miyazawa T, Nashimoto M, Takahashi M.
  • Journal Title: Leuk Res Volume: 37 Issue: 5 Pages: 580-585
  • DOI: 10.1016/j.leukres.2013.01.015
  • Peer Reviewed
• [Journal Article] A naked RNA heptamer targeting the human Bc1-2 mRNA induces apoptosis of HL60 leukemia cells. (2013)
  • Author(s): Masavuki Takahashi, Reyad A. Elbarbry, Aiko Nakashima, Mayumi Abe, Norihiro Watanabe, Miwako Narita, Masuhiro Takahashi, Masato Tamura, Tetsuo Yoshida, Masayuki Nashimoto.
  • Journal Title: Cancer Letters Volume: 328 Issue: 2 Pages: 362-368
  • DOI: 10.1016/j.canlet.2012.10.016
  • Peer Reviewed
• [Journal Article] Direct effect of dasatinib on proliferation and cytotoxicity of natural killer cells in in vitro study (2013)
  • Author(s): Uchiyama T, Sato N, Narita M, Yamahira A, Iwabuchi M, Furukawa T, Sone H, Takahashi M
  • Journal Title: Hematol Oncol Volume: 31 Issue: 3 Pages: 156-63
  • DOI: 10.1002/hon.2034
  • Peer Reviewed
• [Journal Article] Quantification of BCR-ABL mRNA in Plasma/Serum of Patients with Chronic Myelogenous Leukemia (2013)
  • Author(s): Narita M, Saito A, Iwabuchi M, Satoh N, Uchiyama T, Yamahira A, Furukawa T, Sone H, Takahashi M
  • Journal Title: Int J Med Sci Volume: 9 Issue: 10 Pages: 901-8
  • DOI: 10.7150/ijms.4655
  • Peer Reviewed
• [Journal Article] Enhancement of antigen presenting ability in the leukemic plasmacytoid dendritic cell line (PMDC05) by lentiviral vector-mediated transduction of CD80 gene. (2012)
  • Author(s): Yamahira A, Narita M, Ishii K, Jayathilake RM, Iwabuchi M, Satoh N, Uchiyama T, Taniguchi T, Hashimoto S, Kasahara N, Faure E, Bogan B, Takizawa J, Sone H, Takahashi M.
  • Journal Title: Leuk Res Volume: 36 Issue: 12 Pages: 1541-1546
  • DOI: 10.1016/j.leukres.2012.09.002
  • Peer Reviewed
• [Presentation] Antigen presenting exosomes secreted by leukemic plasmacytoid dendritic cell line. (2015)
  • Author(s): Takahashi Masuhiro, Uchiyama Takayoshi, Iwaya Shunpei, Nishizawa Yoshinori, Oiwa Eri, Narita Miwako
  • Organizer: The 18th ECCO & 40th ESMO European Cancer Congress(ECC2015)
  • Place of Presentation: Vienna, Austria
  • Year and Date: 2015-09-25
  • Int'l Joint Research
• [Presentation] 白血病性形質細胞様樹状細胞株PMDC11由来エクソゾームを用いた抗原特異的細胞傷害性T細胞の誘導 (2015)
  • Author(s): 成田美和子、内山孝由、橋本誠雄、瀧澤 淳、高橋益廣
  • Organizer: 第25回樹状細胞研究会
  • Place of Presentation: 岡山コンベンションセンター（岡山県、岡山市）
  • Year and Date: 2015-07-10
• [Presentation] Leukemic plasmacytoid dendritic cell line transduced with caTLR4 gene as a potent antigen presenting cells for immunotherapy. (2015)
  • Author(s): Takahashi Masuhiro, Uchiyama Takayoshi, Iwaya Shunpei, Oiwa Eri, Nishizawa Yoshinori, Hashimoto Shigeo, Bonehill Aude, Kasahara Noriyuki, Narita Miwako
  • Organizer: International Sciety of Cellular Therapy 21th Annual Meeting
  • Place of Presentation: Las Vegas, USA
  • Year and Date: 2015-05-27
  • Int'l Joint Research
• [Presentation] Identification of exosomes secreted by plasmacytoid dendritic cell line (PMDC11) and generation of antigen specific cytotoxic T lymphocytes (CTLs) by the stimulation with the exosomes. (2014)
  • Author(s): Uchiyama T, Narita M, Iwaya S, Nishizawa Y, Oiwa E, Takahashi M
  • Organizer: 日本免疫学会
  • Place of Presentation: 京都
  • Year and Date: 2014-12-10 – 2014-12-12
• [Presentation] caTLR4 gene-transduced leukemic plasmacytoid dendritic cell line (PMDC11) as a potent antigen presenting cells for immunotherapy. (2014)
  • Author(s): Takahashi M, Iwabuchi M, Uchiyama T, Iwaya S, Oiwa E, Nishizawa Y, Hashimoto S, Aude B, Kasahara N, Narita M
  • Organizer: 樹状細胞シンポジウム
  • Place of Presentation: ツール、フランス
  • Year and Date: 2014-09-14 – 2014-09-18
• [Presentation] Potentiation of antigen-specific CTL generation by caTLR4 gene transduction into leukemic plasmacytoid dendritic cell line (PMDC11). (2014)
  • Author(s): Takahashi M, Iwabuchi M, Satoh N, Uchiyama T, Iwaya S, Oiwa E, Nishizawa Y, Hashimoto S, Bogan B, Faure-Kumar E, Kasahara N, Narita M
  • Organizer: アメリカ免疫学会
  • Place of Presentation: ピッツバーグ
  • Year and Date: 2014-05-02 – 2014-05-06
• [Presentation] Analysis of the usage of human T cell receptor b chain variable regions of antigen specific cytotoxic T lymphocytes (2013)
  • Author(s): Uchiyama T, Narita M, Oiwa E, Iwabuchi M, Satoh N, Iwaya S, Nishizawa Y, Takahashi M
  • Organizer: 第42回日本免疫学会
  • Place of Presentation: 幕張
• [Presentation] Transduction of caTLR4 gene enhances antigen-specific CTL inducing ability of leukemic plasmacytoid dendritic cell line (PMDC11) (2013)
  • Author(s): Iwabuchi M, Narita M, Oiwa E, Satoh N, Uchiyama T, Hashimoto S, Bonehill A, Faure-Kumar E, Kasahara N, Takahashi M
  • Organizer: 第42回日本免疫学会
  • Place of Presentation: 幕張
• [Presentation] Ustekinumab improves psoriasis without suppressing tumor antigen-specific cytotoxic T lymphocytes (2013)
  • Author(s): Nishizawa Y, Narita M, Iwaya S, Oiwa E, Iwabuchi M, Satoh N, Uchiyama T, Matsuyama A, Fujiwara H, Takahashi M
  • Organizer: 第42回日本免疫学会
  • Place of Presentation: 幕張
• [Presentation] Identification of IFN-γ production and cytotoxicity in WT1 tetramer+ T cells generated by mixed lymphocyte peptide culture in WT1 peptide immunized persons (2013)
  • Author(s): Oiwa E, Narita M, Iwaya S, Nishizawa Y, Uchiyama T, Iwabuchi M, Satoh N, Takahashi M
  • Organizer: 第42回日本免疫学会
  • Place of Presentation: 幕張
• [Presentation] Successful treatment of molecularly exacerbated CML with WT1 peptides --- association with long-term generation of WT1 tetramer+ T cells (2013)
  • Author(s): Iwaya S, Narita M, Masuko M, Nishizawa Y, Ida T, Oiwa E, Shibazaki Y, Iwabuchi M, Satoh N, Uchiyama T, Sone H, Takahashi M
  • Organizer: 第42回日本免疫学会
  • Place of Presentation: 幕張
• [Presentation] Enhancement of antigen presenting ability in the leukemic plasmacytoid dendritic cell line (PMDC05) by lentiviral vector-mediated transduction of CD80 gene (2012)
  • Author(s): Yamahira A, Narita M, Ishii K, Iwabuchi M, Satoh N, Uchiyama T, Taniguchi T, Hashimoto S, Kasahara N, Faure-Kumar E, Takahashi M
  • Organizer: オーストラリア・ニュージーランド血液学会
  • Place of Presentation: シドニー（オーストラリア）
• [Presentation] Effectiveness of WT1 peptide vaccination in combination with imatinib therapy for an imatinib-resistant CML patient. (2012)
  • Author(s): Narita M, Masuko M, Yamahira A,Uchiyama T, Iwabuchi M, Satoh N, Ida T, Kitajima A, Kitajima T, Furukawa T, Takahashi M
  • Organizer: 第74回日本血液学会総会
  • Place of Presentation: 京都
• [Presentation] Establishment of potent antigen presenting cell line by lentiviral vector-mediated transduction of caTLR4 gene into leukemic plasmacytoid dendritic cell line (PMDC11) (2012)
  • Author(s): Takahashi M, Iwabuchi M, Yamahira A, Satoh N, Uchiyama T, Hashimoto S, Bonehill A, Faure-Kumar E, Bogan B, Kasahara N, Takizawa J, Sone H, Narita M
  • Organizer: 第12回国際DCシンポジウム
  • Place of Presentation: 大邱（韓国）
• [Presentation] Long-lasting Suppression of bcr-abl with Amplification of CTLs in a WT1 Peptide-treated CML Patient (2012)
  • Author(s): Takahashi M, Narita M, Yamahira A, Masuko M, Furukawa T
  • Organizer: 世界生物医学検査会議（WCBLS)
  • Place of Presentation: ベルリン（ドイツ）
• [Presentation] Effectiveness of WT1 peptide vaccination in combination with imatinib therapy for an imatinib-resistant CML patient. (2012)
  • Author(s): Narita M, Masuko M, Yamahira A, Ida T, Kitajima A, Kitajima T, Takai K, Koike T, Takahashi M
  • Organizer: 第17回欧州血液学会
  • Place of Presentation: アムステルダム（オランダ）
• [Remarks] 高橋益廣ホームページ
  • URL: http://www.clg.niigata-u.ac.jp/~matak/index.htm
• [Patent(Industrial Property Rights)] 国際特許公開 (2012)
  • Inventor(s): 梨本正之、高橋益廣、成田美和子
  • Industrial Property Rights Holder: 梨本正之、高橋益廣、成田美和子
  • Industrial Property Rights Type: 特許
  • Filing Date: 2012-08-29
  • Overseas