「希少癌」骨軟部腫瘍のチロシンキナーゼ遺伝子転写変異体の探索

2016 Fiscal Year Research-status Report

• Project/Area Number: 16K15670
• Research Institution: Juntendo University
• Principal Investigator: 末原 義之 順天堂大学, 医学部, 准教授 (70509405)
• Co-Investigator(Kenkyū-buntansha): 高阪 真路 東京大学, 大学院医学系研究科(医学部), 特任助教 (00627119) ; 小林 英介 国立研究開発法人国立がん研究センター, 中央病院, 医員 (40365292)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Keywords: 骨軟部腫瘍 / 転写変異体

Outline of Annual Research Achievements

悪性骨軟部腫瘍の治療成績は、５年生存率が約50％以下と依然不良であり、これは「希少癌」であるが故のデータ量不足や消極的な創薬開発が影響している。対してメジャーな癌、特に肺癌ではEGFR変異などの発生・悪性度因子（driver-oncogene）を治療標的としたTKの阻害剤治療が盛んであり、治療成績は良好である。現在も全癌種・横断的に新規分子標的治療標的（特にTK治療標的）と成り得るdriver-oncogeneが探索され、癌治療のbreakthroughと位置づけられている。
本研究は、難治性かつ「希少癌」である骨軟部腫瘍の生命予後改善の要となる新規分子治療標的の開発を行う。 具体的には転写変異体について、骨軟部腫瘍の手術検体を用いて全Tyrosine Kinase (TK)遺伝子の探索を進める 。その方法として独自のNanostringの手法に基づいたTK転写変異体探索システムを用い、それら同定TK遺伝子転 写変異体については、骨軟部腫瘍における癌化機能の解明とそのターゲットとなるTK阻害剤(TKI)奏功性の検証 を進め、TKIの適応拡大を含めた新規分子治療標的の開発を目指す。
(A)NanoStringシステムによるTK遺伝子転写変異体の探索:全TK遺伝子 転写変異体探索のために、カスタムプローブデザインを行った。解析する骨軟部腫瘍の手術検体よりRNAを抽出しその品質確認を行った。テストサンプルを用いて、nCounter Prep StationによるHybridize, Purify, Immobilize、Digital Analyzerによるプローブに一致したmRNA発現プロファイリング獲得に成功した。それらプロフィリングを用いて統計学的手法を用いてTK遺伝子転写変異体の解析を行った。骨軟部腫瘍検体を用いた解析を行った。
(B)TK遺伝子転写変異体のシークエンスと発現の検証:
テストサンプルにて同定TK遺伝子転写変異体候補に対して、シークエンス解析で転写変異体を同定を行った。骨軟部腫瘍検体を用いた解析を行った。

Current Status of Research Progress

3: Progress in research has been slightly delayed.

Reason

骨軟部腫瘍検体の確保に対して、他施設倫理申請の遅れで、研究計画の遅れが生じた。

Strategy for Future Research Activity

(A)NanoStringシステムによるTK遺伝子転写変異体の探索:
順次解析する骨軟部腫瘍の手術検体よりRNAを抽出しその品質確認を行う。
nCounter Prep StationによるHybridize, Purify, Immobilize、Digital Analyzerによるプローブに一致したmRNA発現プロファイリング獲得を進め、それらプロフィリングを用いて統計学的手法を用いてTK遺伝子転写変異体の解析を行う。引き続き検体・組織型を追加し行う。
骨軟部腫瘍検体を用いた解析を行った。
(B)TK遺伝子転写変異体のシークエンスと発現の検証:
骨軟部腫瘍検体の同定TK遺伝子転写変異体候補に対して、シークエンス解析で転写変異体を同定行う。引き続き検体・組織型を追加し行う。

Causes of Carryover

骨軟部腫瘍検体の確保に対して、他施設倫理申請の遅れで、研究計画の遅れが生じた。

Expenditure Plan for Carryover Budget

(A)NanoStringシステムによるTK遺伝子転写変異体の探索と(B)TK遺伝子転写変異体のシークエンスと発現の検証を順次予定検体・組織型に対して行う。

Research Products

• [Int'l Joint Research] Memorial Sloan-Kettering Cancer Center(米国)
  • Country Name: U.S.A.
  • Counterpart Institution: Memorial Sloan-Kettering Cancer Center
• [Journal Article] TERT promoter mutations are rare in bone and soft tissue sarcomas of Japanese patients. (2016)
  • Author(s): Saito T, Akaike K, Kurisaki-Arakawa A, Toda-Ishii M, Mukaihara K, Suehara Y, Takagi T, Kaneko K, Yao T.
  • Journal Title: Molecular and Clinical Oncology. Volume: 4(1) Pages: 61-64
  • DOI: 10.3892/mco.2015.674
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Protein expression profiling of giant cell tumors of bone treated with denosumab. (2016)
  • Author(s): Mukaihara K, Suehara Y, Kohsaka S, Kubota D, Akaike K, Ishii M, Fujimura T, Okubo T, Takagi T, Yao T, Kaneko K, Ladanyi M, Saito T.
  • Journal Title: PLoS ONE. Volume: 11(2) Pages: e0148401.
  • DOI: http://dx.doi.org/10.1371/journal.pone.0148401
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Expression of F-actin-capping protein subunit beta is associated with cell growth and motility in epithelioid sarcoma. (2016)
  • Author(s): Mukaihara K, Suehara Y, Kohsaka S, Kubota D, Toda-Ishii M, Akaike K, Fujimura T, Kobayashi E, Yao T, Ladanyi, M, Kaneko K, Saito T.
  • Journal Title: BMC cancer. Volume: 16(1) Pages: 206.
  • DOI: 10.1186/s12885-016-2235-z
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] An Outpatient-based Survey on the Recognition of Locomotive Syndrome in Japan: The Results of three Years of Surveys. (2016)
  • Author(s): Tanabe Y, Suehara Y, Okubo T, Kim Y, Ishii M,Kawasaki T, Matsuoka K, Akaike K, Mukaihara K, Kubota D, Okubo N, Maruyama Y, Saito T, Kaneko K.
  • Journal Title: British Journal of Medicine & Medical Research. Volume: 10(9) Pages: 1-9
  • DOI: 10.9734/BJMMR/2016/26127
  • Peer Reviewed / Open Access
• [Journal Article] Paget's sarcoma with sarcoma-specific TP53 mutation arising from a Japanese patient. (2016)
  • Author(s): Akaike K, Toda-Ishii M, Suehara Y, Takagi T, Kaneko K,Yao T, Saito T.
  • Journal Title: International journal of clinical and experimental pathology. Volume: 9(3) Pages: 3978-3986
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Large skin ulcer due to a subcutaneous orthopedic implant after bevacizumab therapy: A case report. (2016)
  • Author(s): Suehara Y, Osawa H, Kubota D, Kato S, Akaike K, Tanabe Y, Saito T, Kaneko K.
  • Journal Title: Bone Joint Surg. Case Connector Volume: 6(3) Pages: e70
  • DOI: https://doi.org/10.2106/JBJS.CC.15.00281
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Clinicopathological effects of protein phosphatase 2, regulatory subunit A, alpha mutations in gastrointestinal stromal tumors (2016)
  • Author(s): Ishii M, Akaike K, Suehara Y, Mukaihara K, Kubota K , Kohsaka S, Okubo T, Mitani K, Mogushi K, Takagi T, Kaneko K, Yao T, Saito T.
  • Journal Title: Modern Pathology. Volume: 29 Pages: 1424-1432
  • DOI: 10.1038/modpathol.2016.138.
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Reduced argininosuccinate synthetase expression in refractory sarcomas. (2016)
  • Author(s): Kim Y, Kobayashi E, Kubota D, Suehara Y, Mukaihara K, Akaike K, Ito A, Kaneko K, Chuman H, Kawai A, Kitano S.
  • Journal Title: Oncotarget. Volume: 7(43) Pages: 70832-70844
  • DOI: 10.18632/oncotarget.12225
  • Peer Reviewed / Open Access
• [Journal Article] A case report of using nivolumab for a malignant melanoma patient with rheumatoid arthritis (2016)
  • Author(s): Kageyama S, Yamaguchi S, Ito S, Suehara Y, Saito T, Akaike K, Miura K, Kato S.
  • Journal Title: Int Canc Conf J. Volume: 5(4) Pages: 192-196
  • DOI: 10.1007/s13691-016-0256-8
  • Peer Reviewed / Open Access
• [Journal Article] A comparative study of 2 screening tools for locomotive syndrome (the “Loco-check” and the “GLFS-25”): An orthopedic outpatient-based survey. (2016)
  • Author(s): Kim Y, Suehara Y, Ishii M, Kawasaki T, Matsuoka K, Okubo T, Okubo N, Tanabe Y, Akaike K, Mukaihara K, Kubota D, Maruyama Y, Saito T, Kaneko K.
  • Journal Title: British Journal of Medicine & Medical Research. Volume: 17(5) Pages: 1-13
  • DOI: 10.9734/BJMMR/2016/28194
  • Peer Reviewed / Open Access
• [Journal Article] A case of secondary chondrosarcoma with TP53 mutation arising from fibrous dysplasia. (2016)
  • Author(s): Tomomasa R, Akaike K, Asahina M, Toda-Ishii M, Yashiro D, Suehara Y, Takagi T, Kaneko K, Yao T, Saito T.
  • Journal Title: International journal of clinical and experimental pathology. Volume: 9(7) Pages: 7230-7236
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] A case of epithelioid hemangioma of the right third rib in a 55-year-old man. (2016)
  • Author(s): Saito T, Akaike K, Tomomasa R, Suehara Y, Takagi T, Kaneko K, Yao T.
  • Journal Title: International journal of clinical and experimental pathology. Volume: 9(9) Pages: 9681-9686
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] Proteomics analysis of characteristic histological changes with Denosumab treatment in giant cell tumors of bone. (2017)
  • Author(s): SueharaY, Mukaihara K, TanabeY, Kohsaka S, Kazuno S, Ishii M, Akaike K, Kubota D, Kaneko K, Saito T.
  • Organizer: the 63th Annual Meeting of the Orthopaedic Research Society
  • Place of Presentation: San Diego, USA
  • Year and Date: 2017-03-19 – 2017-03-22
  • Int'l Joint Research
• [Presentation] Proteomic approaches reveal that inhibition of the IRE1/XBP1 pathway reduces tumor growth in Ewing's sarcoma. (2017)
  • Author(s): Tanabe Y, Suehara Y, Kohsaka S, Mukaihara K, Kubota D, Akaike K, Ishii M, Kazuno S, Mineki R, Kaneko K, Ladanyi M, Saito T.
  • Organizer: the 63th Annual Meeting of the Orthopaedic Research Society
  • Place of Presentation: San Diego, USA
  • Year and Date: 2017-03-19 – 2017-03-22
  • Int'l Joint Research
• [Presentation] Microsatellite instability analysis using the Promega panel in lung adenocarcinoma. (2016)
  • Author(s): Takamochi K, Takahashi F, Suehara Y, Kitano S, Sato E, Kohsaka S, Hayashi T, Mano H, Suzuki K.
  • Organizer: Chest Annual Meeting 2016
  • Place of Presentation: Los Angels, USA
  • Year and Date: 2016-10-22 – 2016-10-26
  • Int'l Joint Research
• [Presentation] The Regulation of Malignant Transformation by PPP2R1A in Alveolar Rhabdomyosarcoma Based On A Proteomic Approach Corresponding To The PAX3/FOXO1 Fusion Gene. (2016)
  • Author(s): Akaike K, Suehara Y, Kohsaka S, Mukaihara K, Ishii M, Kubota D, Tanabe Y, Kazuno S, Mineki R, Fujimura T, Kaneko K, Ladanyi M, Saito T.
  • Organizer: Connective Tissue Oncology Society, 21th Annual Meeting,
  • Place of Presentation: Lisbon, Portugal
  • Year and Date: 2016-09-09 – 2016-09-12
  • Int'l Joint Research
• [Presentation] Protein Expression Profiles Corresponding To Histological Changes With Denosumab Treatment in Giant Cell Tumors of Bone. (2016)
  • Author(s): Suehara Y, Mukaihara K, Tanabe Y, Kohsaka S, Kazuno S, Ishii M, Akaike K, Kubota D, Kaneko K, Saito T.
  • Organizer: Connective Tissue Oncology Society, 21th Annual Meeting,
  • Place of Presentation: Lisbon, Portugal
  • Year and Date: 2016-09-09 – 2016-09-12
  • Int'l Joint Research
• [Presentation] Proteomic approaches revealed inhibition of IRE1/XBP1 pathway reduces tumor growths in Ewing's sarcomas. (2016)
  • Author(s): Tanabe Y, Suehara Y, Kohsaka S, Mukaihara K, Kubota D, Akaike K, Ishii M, Kazuno S, Mineki R, Kaneko K, Ladanyi M, Saito T.
  • Organizer: Connective Tissue Oncology Society, 21th Annual Meeting,
  • Place of Presentation: Lisbon, Portugal
  • Year and Date: 2016-09-09 – 2016-09-12
  • Int'l Joint Research
• [Presentation] Protein expression profiling corresponding to giant cell tumors of bone treated with denosumab. (2016)
  • Author(s): Mukaihara K, Suehara Y, Akaike K, Tanabe Y, Kubota D, Ishii M, Kaneko K, Saito T.
  • Organizer: The 11th Meeting of The Asia Pacific Musculoskeletal Tumour Society (APMSTS)
  • Place of Presentation: Singapore, USA
  • Year and Date: 2016-04-21 – 2016-04-23
  • Int'l Joint Research
• [Presentation] Our efforts to bone and soft tissue tumors and bone metastases at Juntendo University. (2016)
  • Author(s): Kaneko K, Suehara Y, Yonji K, Takagi T.
  • Organizer: The Korean Bone and Joint Tumor Society (KBJTS)
  • Place of Presentation: Incheon, Korea
  • Year and Date: 2016-04-15 – 2016-04-15
  • Int'l Joint Research / Invited