Analysis of human sarcoma stem cells and memory T stem cells

2016 Fiscal Year Annual Research Report

• Project/Area Number: 16H05451
• Research Institution: Sapporo Medical University
• Principal Investigator: 塚原 智英 札幌医科大学, 医学部, 准教授 (20404634)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Keywords: 肉腫幹細胞 / 免疫記憶幹細胞 / 骨肉腫

Outline of Annual Research Achievements

肉腫幹細胞特異抗原の同定を目指して，はじめに新規骨肉腫細胞株OS13を樹立した．OS13は増殖能が高く，またin vitroおよびin vivoにおいて極めて高い造腫瘍能を示した．そこでOS13を限界希釈法によりクローン化し，増殖速度の早いクローン(High clone)と増殖速度の遅いクローン(Low clone)を樹立した．High cloneはLow cloneに比較して，in vitroおよびin vivoにおいて高い造腫瘍能を示した．そこでHigh cloneとLow cloneを用いて次世代シーケンサーを用いた網羅的トランスクリプトーム解析を行った．そしてHigh cloneにおいてmRNAレベルで高発現し，low cloneで発現が低く，かつ精巣胎盤を除く正常組織で極めて発現レベルの低い肉腫幹細胞特異抗原を複数同定した．

ヒト末梢血より薬剤耐性，分化能，高い増殖能を持つ新しい免疫記憶T幹細胞としてyoung memory T（TYM）細胞を同定した．TYM細胞は健常者において，坑ウイルス免疫および坑腫瘍免疫を記憶していることがわかった．そして肉腫患者におけるTYMの頻度は健常者より低かった．肉腫患者においてTYMはエフェクターメモリー細胞やセントラルメモリー細胞と同様にウイルス抗原および腫瘍抗原に対する免疫記憶を保持していた．特に化学療法を受けた肉腫患者においてTYMはセントラルメモリー細胞と比べて，腫瘍抗原に対する免疫記憶を高く保持していた．一方，エフェクター細胞における免疫記憶は低かった．

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

肉腫幹細胞抗原候補が複数同定された．また化学療法後の患者において免疫記憶T幹細胞の免疫記憶を解析できた．

Strategy for Future Research Activity

今後は肉腫幹細胞抗原の機能解析を行って，抗原による肉腫幹細胞の造腫瘍能の制御機構を解明する．また免疫記憶T幹細胞の網羅的遺伝子解析を進める．

Research Products

• [Journal Article] 次世代がんペプチドワクチン療法の開発 (2017)
  • Author(s): 塚原智英，廣橋良彦，鳥越俊彦.
  • Journal Title: 日本臨牀 Volume: 75 Pages: 275-279
• [Journal Article] がん幹細胞の単離と免疫治療に向けた標的抗原の同定． (2017)
  • Author(s): 27.宮本昇，金関貴幸，廣橋良彦，塚原智英，菊池泰弘，佐藤昇志，鳥越俊彦．
  • Journal Title: 日本臨床免疫学会雑誌 Volume: 17 Pages: 40-47
• [Journal Article] Cancer-associated oxidoreductase ERO1-α promotes immune escape through up-regulation of PD-L1 in human breast cancer. (2017)
  • Author(s): Tanaka T, Kutomi G, Kajiwara T, Kukita K, Kochin V, Kanaseki T, Tsukahara T, Hirohashi Y, Torigoe T, Okamoto Y, Hirata K, Sato N, Tamura Y.
  • Journal Title: Oncotarget Volume: 2016 Pages: 未定
  • DOI: 10.18632/oncotarget.14960.
  • Peer Reviewed / Open Access
• [Journal Article] The future of immunotherapy for sarcoma. (2016)
  • Author(s): Tsukahara T , Emori M, Murata K, Mizushima E, Shibayama Y, Kubo T, Kanaseki T, Hirohashi Y, Yamashita T, Sato N, Torigoe T.
  • Journal Title: Expert Opin Biol Ther Volume: 16 Pages: 1049-1057
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Peptide vaccination therapy: Towards the next generation (2016)
  • Author(s): Tsukahara T , Hirohashi Y, Kanaseki T, Nakatsugawa M, Kubo T, Sato N, Torigoe T. Peptide vaccination therapy: Towards the next generation.
  • Journal Title: Pathol Int Volume: 66 Pages: 547-553
  • Peer Reviewed / Acknowledgement Compliant
• [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, 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 Pages: e1165376
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Cancer-associated oxidoreductase ERO1-α drives the production of VEGF via oxidative protein folding and regulating the mRNA level (2016)
  • Author(s): Tanaka T, Kutomi G, Kajiwara T, Kukita K, Kochin V, Kanaseki T, Tsukahara T, Hirohashi Y, Torigoe T, Okamoto Y, Hirata K, Sato N, Tamura Y.
  • Journal Title: Br J Cancer Volume: 114 Pages: 1227-1234
  • Peer Reviewed
• [Journal Article] Establishment and Analysis of Cancer Stem-Like and Non-Cancer Stem-Like Clone Cells from the Human Colon Cancer Cell Line SW480. (2016)
  • Author(s): Takaya A, Hirohashi Y, Murai A, Morita R, Saijo H, Yamamoto E, Kubo T, Nakatsugawa M, Kanaseki T, Tsukahara T, Tamura Y, Takemasa I, Kondo T, Sato N, Torigoe T
  • Journal Title: PLoS One Volume: 14 Pages: :e0158903
  • DOI: 10.1371/journal.pone.0158903. eCollection 2016.
  • Peer Reviewed / Open Access
• [Journal Article] Plasticity of lung cancer stem-like cells is regulated by the transcription factor HOXA5 that is induced by oxidative stress. (2016)
  • Author(s): Saijo H, Hirohashi Y, Torigoe T, Horibe R, Takaya A, Murai A, Kubo T, Kajiwara T, Tanaka T, Shionoya Y, Yamamoto E, Maruyama R, Nakatsugawa M, Kanaseki T, Tsukahara T, Tamura Y, Sasaki Y, Tokino T, Suzuki H, Kondo T, Takahashi H, Sato N
  • Journal Title: Oncotarget Volume: 7 Pages: 50043-50056
  • DOI: 10.18632/oncotarget.10571
  • Peer Reviewed / Open Access
• [Journal Article] Hypoxia augments MHC class I antigen presentation via facilitation of ERO1-α-mediated oxidative folding in murine tumor cells. (2016)
  • Author(s): Kajiwara T, Tanaka T, Kukita K, Kutomi G, Saito K, Okuya K, Takaya A, Kochin V, Kanaseki T, Tsukahara T, Hirohashi Y, Torigoe T, Hirata K, Sato N, Tamura Y.
  • Journal Title: Eur J Immunol. Volume: 46 Pages: 2842-2851
  • Peer Reviewed
• [Presentation] Identification of a human novel memory T cell population with the characteristics of chemo-resistance. (2016)
  • Author(s): 塚原 智英
  • Organizer: 第20回日本がん免疫学会総会
  • Place of Presentation: 大阪国際交流センター（大阪市）
  • Year and Date: 2016-07-28 – 2016-07-28
• [Book] Osteosarcoma - A Multidisciplinary Approach to Treatment (2016)
  • Author(s): 5.Tsukahara T, Wada T. Immunotherapy for osteosarcoma
  • Total Pages: 270
  • Publisher: Springer Japan KK