Immune response against sarcoma stem-like cell antigen.

• Project/Area Number: 25462344
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Orthopaedic surgery
• Research Institution: Sapporo Medical University
• Principal Investigator: TSUKAHARA Tomohide 札幌医科大学, 医学部, 講師 (20404634)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: sarcoma stem cell / CTL clone / immunotherapy / 肉腫幹細胞 / CTL / 腫瘍抗原 / 免疫療法 / 自家CTLクローン / 抗原 / SP / 人工CTL

Outline of Final Research Achievements

We isolated sarcoma stem-like cells (SSC) from the malignant fibrous histiocytoma cell line, MFH2003. After autologous PBMC was stimulated with SSC, we established CTL clone Tc4c-6 recognizing SSC in the context of HLA class I. Next, we isolated cDNA of T cell receptor (TCR) alpha (A) & beta (B) chains. Two TCRAs were isolated (A5 and A7). Each cDNA of TCRA5B and TCRA7B was transfected into 58a-b- cells lacking TCR (58A5B and 58A7B). 58A7B cells could specifically recognize SSCs, however, the response was weak. Next, we transfected TCRA7B into the Jurkat cell line expressing NFAT-luciferase (Jrt/MA). The resultant cell line (J-A7B) recognized autologous SSCs but not non-SSCs. Moreover, J-A7B also recognized allogeneic osteosarcoma cell line KIKU in the context of HLA class I. Now we are trying to determine the HLA class I allele that presents the CTL epitope peptide derived from SSC antigen. These findings are useful for cDNA expression cloning of autologous CTL-defined SSC antigen.

Research Products

• [Journal Article] Identification of a novel human memory T cell population with the characteristics of stem-like chemo-resistance. (2016)
  • Author(s): Murata K, Tsukahara T (corresponding author), Emori M, Shibayama Y, Mizushima E, Matsumiya H, Yamashita K, Kaya M, Hirohashi Y, Kanaseki T, Kubo T, Himi T, Ichimiya S, Yamashita T, Sato N, Torigoe T.
  • Journal Title: OncoImmunology Volume: 5 Issue: 6 Pages: e1165376-e1165376
  • DOI: 10.1080/2162402x.2016.1165376
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Prognostic impact of CD109 expression in myxofibrosarcoma. (2015)
  • Author(s): Emori M, Tsukahara T, Murata K, Sugita S, Sonoda T, Kaya M, Soma T, Sasaki M, Nagoya S, Hasegawa T, Wada T, Sato N, Yamashita T
  • Journal Title: J Surg Oncol. Volume: 111 Issue: 8 Pages: 975-979
  • DOI: 10.1002/jso.23934
  • Peer Reviewed
• [Journal Article] Heat shock protein DNAJB8 is a novel target for immunotherapy of colon cancer-initiating cells. (2014)
  • Author(s): Morita R, Nishizawa S, Torigoe T, Takahashi A, Tamura Y, Tsukahara T, Kanaseki T, Sokolovskaya A, Kochin V, Kondo T, Hashino S, Asaka M, Hara I, Hirohashi Y, Sato N.
  • Journal Title: Cancer Sci Volume: 105 Issue: 4 Pages: 389-395
  • DOI: 10.1111/cas.12362
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] High Expression of CD109 Antigen Regulates the Phenotype of Caneer Stem-Like Cells/Cancer-lnitiating Cells in the Novel Epithelioid Sarcoma Cell Line ESX and Is Related to Poor Prognosis of Soft Tissue Sarcoma. (2013)
  • Author(s): Emori M, Tsukahara T, Murase M, Kano M, Murata K, Takahashi A, Kubo T, Asanuma H, Yasuda K, Kochin V, Kaya M, Nagoya S, Nishio J, Iwasaki H, Sonoda T, Hasegawa T, Torigoe T, Wada T, Yamashita T, Sato N.
  • Journal Title: PLoS One. Volume: 8 Issue: 12 Pages: e84187-e84187
  • DOI: 10.1371/journal.pone.0084187
  • Peer Reviewed
• [Journal Article] Ectopically expressed variant form of sperm mitochondria-associated cysteine-rich protein augments tumorigenicity of the stem cell population of lung adenocarcinoma cells. (2013)
  • Author(s): Takahashi A, Hirohashi Y, Torigoe T, Tamura Y, Tsukahara T, Kanaseki T, Kochin V, Saijo H, Kubo T, Nakatsugawa M, Asanuma H, Hasegawa T, Kondo T, Sato N.
  • Journal Title: PLoS One Volume: 8 Issue: 11 Pages: e69095-e69095
  • DOI: 10.1371/journal.pone.0069095
  • Peer Reviewed
• [Journal Article] Small proline-rich protein-1B is overexpressed in human oral squamous cell cancer stem-like cells and is related to their growth through activation of MAP kinase signal. (2013)
  • Author(s): Michifuri Y, Hirohashi Y, Torigoe T, Miyazaki A, Fujino J, Tamura Y, Tsukahara T, Kanaseki T, Kobayashi J, Sasaki T, Takahashi A, Nakamori K, Yamaguchi A, Hiratsuka H, Sato N.
  • Journal Title: Biochem Biophys Res Commun Volume: 439 Issue: 1 Pages: 96-102
  • DOI: 10.1016/j.bbrc.2013.08.021
  • Peer Reviewed
• [Presentation] ヒト骨肉腫の分子病理と免疫応答 (2015)
  • Author(s): 塚原 智英
  • Organizer: 第61回日本病理学会秋期特別総会
  • Place of Presentation: 東京大学安田講堂（東京都文京区）
  • Year and Date: 2015-11-05
• [Presentation] 骨軟部肉腫に対するペプチドワクチン療法 (2015)
  • Author(s): 塚原 智英
  • Organizer: 第43回日本臨床免疫学会
  • Place of Presentation: 神戸国際会議場（神戸市）
  • Year and Date: 2015-10-22
• [Presentation] Development of the functional probe against an autologous CTL-defined sarcoma stem cell-associated antigen (2013)
  • Author(s): 塚原智英
  • Organizer: 日本癌学会学術総会
  • Place of Presentation: 横浜
• [Presentation] Development of peptide vaccination therapy targeting apoptosis regulator protein PBF for patients with osteosarcoma. (2013)
  • Author(s): 塚原智英
  • Organizer: International Congress of Immunology
  • Place of Presentation: Milan, Italy
• [Presentation] Co-existence of HLA class I expression and CD8+ T cell infiltration is associated with favorable clinical outcomes in Ewing’s sarcoma family of tumors (ESFT). (2013)
  • Author(s): Yabe H, Tsukahara T, Kawaguchi S, Torigoe T, Sato N, Morioka H, Yabe H.
  • Organizer: International Congress of Immunology
  • Place of Presentation: Milan, Italy
• [Book] Osteosarcoma - A Multidisciplinary Approach to Treatment (2016)
  • Author(s): Tsukahara T, Wada T
  • Total Pages: 270
  • Publisher: Springer Japan KK
• [Book] The Frontiers in Life Sciences『がんと免疫』 (2015)
  • Author(s): 3.塚原智英，鳥越俊彦，廣橋良彦，金関貴幸，Kochin Vitaly，佐藤昇志．
  • Total Pages: 192
  • Publisher: 南山堂
• [Book] Inflammation and immunity in cancer (2015)
  • Author(s): Sato N, Hirohashi Y, Tsukahara T, Kanaseki T, Kochin V, Tamura Y, Torioge T
  • Total Pages: 268
  • Publisher: Springer Japan KK
• [Patent(Industrial Property Rights)] Ｔ細胞レセプターとその利用 (2016)
  • Inventor(s): 鳥越俊彦，田路真悟、中野一絵、齊藤尚吾，塚原智英，廣橋良彦
  • Industrial Property Rights Holder: 鳥越俊彦，田路真悟、中野一絵、齊藤尚吾，塚原智英，廣橋良彦
  • Industrial Property Rights Type: 特許
  • Filing Date: 2016-03-18
• [Patent(Industrial Property Rights)] 骨肉腫特異抗原PBF由来A11拘束性CTLエピトープ (2015)
  • Inventor(s): 李棟梁，中野一絵，田路真悟，塚原智英，鳥越俊彦，佐藤昇志
  • Industrial Property Rights Holder: 李棟梁，中野一絵，田路真悟，塚原智英，鳥越俊彦，佐藤昇志
  • Industrial Property Rights Type: 特許
  • Filing Date: 2015-11-20