蛍光イメージングを用いた悪性骨軟部腫瘍の転移機序の解明と新規治療法の開発

2014 Fiscal Year Annual Research Report

• Project/Area Number: 26293333
• Research Institution: Kanazawa University
• Principal Investigator: 土屋 弘行 金沢大学, 医学系, 教授 (40227434)
• Co-Investigator(Kenkyū-buntansha): 山本 憲男 金沢大学, 医学保健学総合研究科, 特任教授 (90332668) ; 林 克洋 金沢大学, 大学病院, 助教 (80507054) ; 木村 浩明 名古屋市立大学, 医学(系)研究科(研究院), 助教 (50608693) [Withdrawn]
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: 骨肉腫 / 骨転移 / 肉腫

Outline of Annual Research Achievements

【腫瘍特異的バクテリア治療】
カリフォルニア大学サンディエゴ校で樹立したA1-Rを用いた。腫瘍モデルとして、乳癌骨転移モデル、軟部肉腫モデルを作成した。乳癌骨転移モデルにはMDA-MB-435を用いた。GFPで標識したMDSA435細胞をヌードマウスの左心室に注射し、骨転移巣からGFP発現細胞を回収、培養して再び左心室に注射するというサイクルを繰り返すことにより高転移株を得た。MDA435高転移株をヌードマウスに注射し、A1-Rで治療をしたところ、転移巣の形成が有意に抑制された。また、MDA435を脛骨に直接注射することにより得られた腫瘍に対し、A1-Rで治療したところ、腫瘍の成長が治療群で有意に低下した（Miwa, etl al. Oncotarget 5: 7119-25, 2014）。ヒト線維肉腫HT1080をヌードマウスの大腿四頭筋に注射することにより軟部肉腫モデルを作成、HT1080をヌードマウスの尾静脈に注射することにより肺転移モデルを作成した。これらのモデルに対してA1-Rで治療したところ、原発モデル、転移モデルのいずれにも抑制効果が見られた（Miwa, et al. Oncotarget 5: 12849-61, 2014）。以上のことから、A1-Rは腫瘍に対する新しい治療として期待できるものと考えた。
【蛍光ガイド下手術】
RFPで標識したヒト骨肉腫細胞株143B、GFPで標識したヒト前立腺癌細胞株PC-3をヌードマウスの脛骨に注射することにより骨肉腫モデル、骨転移モデルを作成した。腫瘍を肉眼で辺縁切除術を行ったグループと、蛍光イメージング装置で蛍光を確認しながら切除術を行ったグループにおける遺残腫瘍、腫瘍の再発率、再発腫瘍の蛍光を比較した。いずれの腫瘍モデルにおいても、FGSは有意に遺残腫瘍、腫瘍の再発を抑制した（Miwa, et al. J Orthop Res 32: 1596-601, 2014; 19.Miwa, et al. J Surg Res 192: 124-33, 2014）。このことから、FGSは腫瘍の確実な腫瘍切除に有用であると考えた。

Current Status of Research Progress

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

Reason

骨転移モデルは脛骨に直接注射する方法と左心室に注射する方法により安定したモデルが得られており、それらのモデルにおいて腫瘍特異的サルモネラが抗腫瘍効果を示すこと、蛍光ガイド下手術により術後再発率が低下するというデータを得ている。

Strategy for Future Research Activity

今回行った新規治療の実験について、その安全性、臨床応用のための実験を行う。骨転移モデルにおける新規プラチナ製剤の効果について実験を進める。また、蛍光ガイド下手術に必要な腫瘍の蛍光標識方法について実験を進める。

Causes of Carryover

現時点で新規プラチナ製剤の治療実験の成果が出ておらず、また、蛍光ガイド下手術についての追加実験も必要と考えている。これらの実験を行うために次年度も研究費が必要である。

Expenditure Plan for Carryover Budget

上記実験に必要なヌードマウス、細胞培養液、蛍光抗体などの試薬を購入する予定である。

Research Products

• [Journal Article] Cell-cycle fate-monitoring distinguishes individual chemosensitive and chemoresistant cancer cells in drug-treated heterogeneous populations demonstrated by real-time FUCCI imaging. (2015)
  • Author(s): Miwa S, Yano S, Kimura H, Yamamoto M, Toneri M, Matsumoto Y, Uehara F, Hiroshima Y, Murakami T, Hayashi K, Yamamoto N, Bouvet M, Fujiwara T, Tsuchiya H, Hoffman RM.
  • Journal Title: Cell Cycle Volume: 14 Pages: 621-629
  • DOI: 10.4161/15384101.2014.991604
  • Peer Reviewed
• [Journal Article] Salvage method for unplanned excision of soft tissue sarcoma: long-term results of second-look surgery following radio-hyperthermo-chemotherapy. (2015)
  • Author(s): Hayashi K, Yamada S, Inatani H, Okamoto H, Takeuchi A, Nishida H, Yamamoto N, Tsuchiya H, Otsuka T.
  • Journal Title: Anticancer Research Volume: 35 Pages: 493-498
  • Peer Reviewed
• [Journal Article] Effectiveness of two novel anionic and cationic platinum complexes in the treatment of osteosarcoma. (2015)
  • Author(s): Igarashi K, Yamamoto N, Hayashi K, Takeuchi A, Miwa S, Odani A, Tsuchiya H.
  • Journal Title: Anticancer Agents Med Chem Volume: 15 Pages: 390-399
  • Peer Reviewed
• [Journal Article] Efficacy of triplet regimen antiemetic therapy for chemotherapy-induced nausea and vomiting (CINV) in bone and soft tissue sarcoma patients receiving highly emetogenic chemotherapy, and an efficacy comparison of single-shot palonosetron and consecutive-day granisetron for CINV in a randomized, single-blinded crossover study. (2015)
  • Author(s): Kimura H, Yamamoto N, Shirai T, Nishida H, Hayashi K, Tanzawa Y, Takeuchi A, Igarashi K, Inatani H, Shimozaki S, Kato T, Aoki Y, Higuchi T, Tsuchiya H.
  • Journal Title: Cancer Med Volume: 4 Pages: 333-341
  • DOI: 10.1002/cam4.373
  • Peer Reviewed / Open Access
• [Journal Article] Surgical site infection after total en bloc spondylectomy: risk factors and the preventive new technology. (2015)
  • Author(s): Hayashi H, Murakami H, Demura S, Kato S, Yoshioka K, Shinmura K, Yokogawa N, Ishii T, Fang X, Shirai T, Tsuchiya H.
  • Journal Title: Spine J Volume: 15 Pages: 132-7
  • Peer Reviewed
• [Journal Article] Inhibition of spontaneous and experimental lung metastasis of soft-tissue sarcoma by tumor-targeting Salmonella typhimurium A1-R (2014)
  • Author(s): Miwa S, Zhang Y, Baek KE, Uehara F, Yano S, Yamamoto M, Hiroshima Y, Matsumoto Y, Kimura H, Hayashi K, Yamamoto N, Tsuchiya H, Bouvet M, Hoffman RM, Zhao M
  • Journal Title: Oncotarget Volume: 5 Pages: 12849-12861
  • Peer Reviewed / Open Access
• [Journal Article] Tumor-targeting Salmonella typhimurium A1-R prevents experimental human breast cancer bone metastasis in nude mice (2014)
  • Author(s): Miwa S, Yano S, Zhang Y, Matsumoto Y, Uehara F, Yamamoto M, Hiroshima Y, Kimura H, Hayashi K, Yamamoto N, Bouvet M, Tsuchiya H, Hoffman RM, Zhao M
  • Journal Title: Oncotarget Volume: 5 Pages: 7119-71125
  • Peer Reviewed / Open Access
• [Journal Article] Fluorescence-guided surgery improves outcome in an orthotopic osteosarcoma nude-mouse model (2014)
  • Author(s): Miwa S, Hiroshima Y, Yano S, Zhang Y, Matsumoto Y, Uehara F, Yamamoto M, Kimura H, Hayashi K, Bouvet M, Tsuchiya H, Hoffman RM
  • Journal Title: Journal of Orthopaedic Ressearch Volume: 32 Pages: 1596-1601
  • DOI: 10.1002/jor.22706
  • Peer Reviewed
• [Journal Article] Fluorescence guided-surgery of prostate cancer bone metastasis (2014)
  • Author(s): Miwa S, Matsumoto Y, Hiroshima Y, Yano S, Uehara F, Yamamoto M, Zhang Y, Kimura H, Hayashi K, Yamamoto N, Bouvet M, Sugimoto N, Tsuchiya H, Hoffman RM
  • Journal Title: Journal of Surgical Research Volume: 192 Pages: 124-133
  • DOI: 10.1016/j.jss.2014.05.049
  • Peer Reviewed
• [Journal Article] Late recurrence of osteosarcoma: a report of two cases. (2014)
  • Author(s): Igarashi K, Yamamoto N, Shirai T, Nishida H, Hayashi K, Tanzawa Y, Kimura H, Takeuchi A, Miwa S, Inatani H, Shimozaki S, Kato T, Tsuchiya H.
  • Journal Title: J Orthop Surg (Hong Kong) Volume: 22 Pages: 415-419
  • Peer Reviewed / Open Access
• [Journal Article] Motor function of the upper-extremity after transection of the second thoracic nerve root during total en bloc spondylectomy. (2014)
  • Author(s): Yokogawa N, Murakami H, Demura S, Kato S, Yoshioka K, Hayashi H, Ishii T, Fujii M, Igarashi T, Tsuchiya H.
  • Journal Title: PLoS One Volume: 9 Pages: e109838
  • DOI: 10.1371/journal.pone.0109838
  • Peer Reviewed / Open Access
• [Journal Article] Outcomes and complications of reconstruction using tumor-bearing frozen autografts in patients with metastatic bone tumors. (2014)
  • Author(s): Miwa S, Takeuchi A, Shirai T, Yamamoto N, Hayashi K, Nishida H, Kimura H, Igarashi K, Tsuchiya H.
  • Journal Title: Anticancer Res Volume: 34 Pages: 5569-5577
  • Peer Reviewed
• [Journal Article] Factors that influence functional outcome after total or subtotal scapulectomy: Japanese Musculoskeletal Oncology Group (JMOG) study. (2014)
  • Author(s): Hayashi K, Iwata S, Ogose A, Kawai A, Ueda T, Otsuka T, Tsuchiya H.
  • Journal Title: PLoS One Volume: 9 Pages: e100119
  • DOI: 10.1371/journal.pone.0100119
  • Peer Reviewed / Open Access
• [Journal Article] Antimicrobial megaprostheses supported with iodine. (2014)
  • Author(s): Shirai T, Tsuchiya H, Nishida H, Yamamoto N, Watanabe K, Nakase J, Terauchi R, Arai Y, Fujiwara H, Kubo T.
  • Journal Title: J Biomater Appl Volume: 29 Pages: 617-23
  • DOI: 10.1177/0885328214539365
  • Peer Reviewed
• [Journal Article] Accurate leg length measurement in total hip arthroplasty: a comparison of computer navigation and a simple manual measurement device. (2014)
  • Author(s): Ogawa K, Kabata T, Maeda T, Kajino Y, Tsuchiya H.
  • Journal Title: Clin Orthop Surg Volume: 6 Pages: 153-8
  • DOI: 10.4055/cios.2014.6.2.153
  • Peer Reviewed / Open Access
• [Journal Article] Perioperative complications of total en bloc spondylectomy: adverse effects of preoperative irradiation. (2014)
  • Author(s): Yokogawa N, Murakami H, Demura S, Kato S, Yoshioka K, Hayashi H, Ishii T, Igarashi T, Fang X, Tsuchiya H.
  • Journal Title: PLoS One Volume: 9 Pages: e98797
  • DOI: 10.1371/journal.pone.0098797
  • Peer Reviewed / Open Access
• [Journal Article] Grafted tendon healing in femoral and tibial tunnels after anterior cruciate ligament reconstruction. (2014)
  • Author(s): Nakase J, Kitaoka K, Toratani T, Kosaka M, Ohashi Y, Tsuchiya H.
  • Journal Title: J Orthop Surg (Hong Kong) Volume: 22 Pages: 65-9
  • Peer Reviewed / Open Access
• [Journal Article] Uncultured autogenous adipose-derived regenerative cells promote bone formation during distraction osteogenesis in rats. (2014)
  • Author(s): Nomura I, Watanabe K, Matsubara H, Hayashi K, Sugimoto N, Tsuchiya H.
  • Journal Title: Clin Orthop Relat Res Volume: 472 Pages: 3798-806
  • DOI: 10.1007/s11999-014-3608-8
  • Peer Reviewed
• [Journal Article] Prevention of pin tract infection with iodine-supported titanium pins. (2014)
  • Author(s): Shirai T, Watanabe K, Matsubara H, Nomura I, Fujiwara H, Arai Y, Ikoma K, Terauchi R, Kubo T, Tsuchiya H.
  • Journal Title: J Orthop Sci Volume: 19 Pages: 598-602
  • DOI: 10.1007/s00776-014-0561-z
  • Peer Reviewed
• [Presentation] Reconstruction of malignant bone tumors using pedicle frozen autograft treated with liquid nitrogen (2014)
  • Author(s): Tsuchiya H
  • Organizer: XXVI SICOT Triennial World Congress
  • Place of Presentation: SulAmerica Convention Center （Rio de Janeiro, Brazil）
  • Year and Date: 2014-11-19 – 2014-11-22
• [Presentation] Advantage of antimicrobial iodine-coated implants for prevention and treatment of implant-associated orthopaedic infections (2014)
  • Author(s): Tuchiya H
  • Organizer: XXVI SICOT Triennial World Congress
  • Place of Presentation: SulAmerica Convention Center （Rio de Janeiro, Brazil）
  • Year and Date: 2014-11-19 – 2014-11-22
• [Presentation] Epiphyseal preservation and biological reconstruction for metaphyseal osteosarcoma (2014)
  • Author(s): Tsuchiya H, Matsubara H, Takeuchi A, Yamamoto N, Nishida H, Hayashi K, Aikawa T, Yoshida Y
  • Organizer: The 24th Annual Meeting of LLRS (ASAMI-North America)
  • Place of Presentation: Fairmont The Queen Elizabeth （Montreal, Canada）
  • Year and Date: 2014-07-25 – 2014-07-26
• [Presentation] 患肢温存手術の進歩—夢の途中 (2014)
  • Author(s): 土屋弘行
  • Organizer: 第87回日本整形外科学会学術総会
  • Place of Presentation: 神戸ポートピアホテル、神戸国際展示場、神戸国際会議場（兵庫県神戸市）
  • Year and Date: 2014-05-21 – 2014-05-25
• [Presentation] Fluorescence-guided surgery decreases residual tumor in primary osteosarcoma model (2014)
  • Author(s): Miwa S, Hiroshima Y, Yano S, Matsumoto Y, Uehara F, Yamamoto M, Kimura H, Hayashi K, Tsuchiya H, Hoffman RM
  • Organizer: 2014 American Society of Cancer Research (AACR) annual meeting
  • Place of Presentation: San Diego Convention Center （San Diego, USA）
  • Year and Date: 2014-04-05 – 2014-04-09
• [Presentation] The efficacy of tumor-targeting Salmonella typhimurium A1-R for bone metastasis (2014)
  • Author(s): Miwa S, Yano S, Zhang Y, Uehara F, Matsumoto Y, Hiroshima Y, Yamamoto M, Kimura H, Hayashi K, Tsuchiya H, Zhao M, Hoffman RM
  • Organizer: 2014 American Society of Cancer Research (AACR) annual meeting
  • Place of Presentation: San Diego Convention Center （San Diego, USA）
  • Year and Date: 2014-04-05 – 2014-04-09