Screening for alternative transcription initiation of tyrosine kinase genes in bone and soft tissue tumors

2018 Fiscal Year Annual Research 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 国立研究開発法人国立がん研究センター, 研究所, 主任研究員 (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, 医長 (40365292)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Keywords: 骨軟部腫瘍 / 遺伝子変異体 / 融合遺伝子 / tyrosine Kinase

Outline of Annual Research Achievements

本研究は、難治性かつ希少がん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った。 具体的にはTyrosine Kinase (TK)遺伝子の融合遺伝子及び転写変異体について、骨軟部腫瘍の手術検体を用いて全TK遺伝子に対してスクリーニング探索を進めた。その方法としてNanostringの手法に基づいたmRNA発現を5’-, 3’-にて観察を行い（検体よりRNAを抽出し、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い、nCounter Prep Stationを用いて、Hybridize, Purify, Immobilizeを進め、Digital Analyzerによりプローブに一致したmRNA発現プロファイリングを獲得し、統 計学的手法を用いて同定を行った）、TK遺伝子の融合遺伝子及び転写変異体候補の選出を行った。それら同定TK遺伝子の融合遺伝子及び転写変異体候補については、その発現・変異検証と癌化機能及び治療標的としての阻害剤奏功性の検証 を進めた。それらを包括的に検証確認を進め新規分子治療標的の開発を行った。対象は骨軟部腫瘍において（紡錘型細胞肉腫、小児腫瘍、inflammatory myofibroblastic tumor(IMT) /Inflammatory pseudotumor(IPT)）に対して探索を進め、ALK（融合遺伝子または転写変異体）、ROS1（融合遺伝子または転写変異体）、NTRK（融合遺伝子または転写変異体）候補の同定に成功し、いずれも検証の結果融合遺伝子あった。具体的には平滑筋肉腫にROS1融合遺伝子を同定、小児腫瘍にNTRK融合遺伝子を同定、またIMT/IPTにALK、ROS1、NTRK融合遺伝子を同定した。それぞれにおいて癌化能及び阻害剤奏効性も確認に成功した。

Research Products

• [Int'l Joint Research] Memorial Sloan Kettering Cancer Center/Department of Pathology/Marc Ladanyi(米国)
  • Country Name: U.S.A.
  • Counterpart Institution: Memorial Sloan Kettering Cancer Center/Department of Pathology/Marc Ladanyi
  • # of Other Institutions: 1
• [Journal Article] Comprehensive assay for the molecular profiling of cancer by using DNA and RNA hybridization capture-based target enrichment from formalin- fixed paraffin-embedded specimens. (2019)
  • Author(s): Kohsaka S, Suehara Y, Mano H et al.
  • Journal Title: Cancer Science Volume: 110 Pages: 1464-1479
  • DOI: 10.1111/cas.13968.
  • Peer Reviewed
• [Journal Article] Multicenter experience with large panel next generation sequencing in patients with advanced solid cancers in Japan (2019)
  • Author(s): Kato S, Hayashi T, Suehara Y, Hananoue H, Yamanaka S, Ichikawa Y, Higurashi T, Ohashi K, Yamaguchi S, Nozaki Y, Terao Y, Saito T, Yao T, Nakajima A, Syed A, Zehir A, Ladanyi M, Kato S.
  • Journal Title: Japanese Journal of Clinical Oncology Volume: 49 Pages: 174-182
  • DOI: 10.1093/jjco/hyy173.
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Squamous cell carcinoma arising from chronic osteomyelitis massively expanding into the medullary cavity: A case report. (2019)
  • Author(s): Kurihara T, Suehara Y, Akaike K, Hayashi T, Okubo T, Kim Y, Takagi T, Kaneko K, Yao T, Saito T.
  • Journal Title: Human Pathology Case reports Volume: 16 Pages: 100289
  • DOI: https://doi.org/10.1016/j.ehpc.2019.01.008
  • Peer Reviewed / Open Access
• [Journal Article] Fibrocartilaginous mesenchymoma of the tibia with predominant microcystic features: A case report and literature review (2019)
  • Author(s): Saito T, Motoi T, Suehara Y, Takagi T, Okubo T, Kurihara T, Hayashi T, Kudo T, Maruyama Y, Kaneko K, YaoT.
  • Journal Title: Human Pathology Case reports Volume: 16 Pages: 100288
  • DOI: https://doi.org/10.1016/j.ehpc.2019.01.007
  • Peer Reviewed / Open Access
• [Journal Article] Proteomic signatures corresponding to the SS18/SSX fusion gene. (2018)
  • Author(s): Ishii M, Suehara Y, Kohsaka S, Tanabe Y, Hayashi T, Kazuno S, Tanabe Y, Akaike K, Mukaihara K, Kim Y, Okubo T, Takamochi K, Takahashi F, Kaneko K, Saito T.
  • Journal Title: Oncotarget Volume: 9 Pages: 37509-37519
  • DOI: 10.18632/oncotarget.26493
  • Peer Reviewed / Open Access
• [Journal Article] PPP2R1A regulated by PAX3/FOXO3 fusion contributes to the acquisition of aggressive behavior in PAX3/FOXO1-positive alveolar rhabdomyosarcoma. (2018)
  • Author(s): Akaike K, Suehara Y, Kohsaka S, Hayashi T, Tanabe Y, Kazuno S, Mukaihara K, Ishii M, Kurihara T, Youngji K, Okubo T, Hayashi Y, Takamochi K, Takahashi F, Kaneko K, Ladanyi M, Saito T.
  • Journal Title: Oncotarget Volume: 9 Pages: 25206-25215
  • DOI: 10.18632/oncotarget.25392
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] A comprehensive clinicopathological analysis of gastric adenocarcinoma with enteroblastic differentiation using next generation sequencing with emphasis on the distinction from socalled hepatoid adenocarcinoma. (2018)
  • Author(s): Akazawa Y, Saito T, Hayashi T, Yanai Y, Tsuyama S, Akaike K,Suehara Y, Takahashi F, Takamochi K, Murakami T, Watanabe S, Yao T.
  • Journal Title: Human Pathology Volume: 78 Pages: 79-88
  • DOI: 10.1016/j.humpath.2018.04.022.
  • Peer Reviewed / Open Access
• [Journal Article] KCTD12 may be negatively regulated by KIT in gastrointestinal stromal tumors. (2018)
  • Author(s): Suehara Y, Akaike K, Mukaihara K, Kurisaki-Arakawa A, Kubota D, Okubo T, Mitomi H, Mitani K, Takahashi M, Toda-Ishii M, Taguchi T, Takagi T, Kaneko K, Yao T, Saito T.
  • Journal Title: Oncotarget Volume: 9 Pages: 27016-27026
  • DOI: 10.18632/oncotarget.25469
  • Peer Reviewed / Open Access
• [Journal Article] IRE1α-XBP1 inhibitors exerted anti-tumor activities in Ewing's sarcomas. (2018)
  • Author(s): Tanabe Y, Suehara Y, Kohsaka S, Hayashi T, Akaike K, Mukaihara K, Kurihara T, Kim Y, Okubo T, Ishii M, Kazuno S, Kaneko K, Saito T.
  • Journal Title: Oncotarget Volume: 9 Pages: 14428-14443
  • DOI: 10.18632/oncotarget.24467.
  • Peer Reviewed / Open Access
• [Presentation] Clinicopathological characteristics of lung adenocarcinoma with EGFR compound mutations. (2019)
  • Author(s): Hayashi T, Kohsaka S, Takamochi K, Takahashi F, Suehara Y, SaitoT, Suzuki K, Mano H, Yao T.
  • Organizer: USCAP, the 2019 Annual Meeting
  • Int'l Joint Research
• [Presentation] Metabolic fluctuations in adriamycin-resistant osteosarcomas using metabolomic approaches (2019)
  • Author(s): Sasa K, Suehara Y, Akaike K, Hayashi T, Taka H, Youngji K, Okubo T, Kaneko K, Saito T.
  • Organizer: the 65th Annual Meeting of the Orthopaedic Research Society
  • Int'l Joint Research
• [Presentation] IRE1α-XBP1 Inhibitor exerts antiyumor activity in osteosarcoma. (2019)
  • Author(s): Kurihara T, Suehara Y, Hayashi T, Akaike K, Okubo T, Kim Y, Kaneko K, Saito T.
  • Organizer: the 65th Annual Meeting of the Orthopaedic Research Society
  • Int'l Joint Research
• [Presentation] Comprehensive mRNA-based screen for tyrosine kinase fusions and a de novo alternative transcription initiation site in soft tissue sarcomas. (2018)
  • Author(s): Suehara Y, Kohsaka S, Kurisaki A, Akaike K, Hayashi T, Mogushi K, Okubo T, Kim Y, Sato S, Kobayashi E, Kaneko K, Mano H, Saito T.
  • Organizer: European Society For Medical Oncology ASIA,
  • Int'l Joint Research
• [Presentation] IRE1α-XBP1 Inhibitor exerts antiyumor activity in osteosarcoma. (2018)
  • Author(s): Kurihara T, Suehara Y, Hayashi T, Akaike K, Okubo T, Kim Y, Kaneko K, Saito T.
  • Organizer: Connective Tissue Oncology Society, 23th Annual Meeting
• [Presentation] Expanded access of larotrectenib to manage pediatric TRK fusion sarcomas. (2018)
  • Author(s): Suehara Y, Trahair T, Kirby M, Saito S, Capra M, Ghosh T, Henry D, Ku NC, Cox MC, Kato S, Ziegler DS.
  • Organizer: 50th Congress of International Society of Paediatric Oncology,