肉腫融合遺伝子による発がんメカニズムの統合的探索

2020 Fiscal Year Annual Research Report

• Project/Area Number: 18H02677
• Research Institution: National Cancer Center Japan
• Principal Investigator: 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, ユニット長 (00627119)
• Co-Investigator(Kenkyū-buntansha): 椨 康一 東京医科歯科大学, 難治疾患研究所, 助教 (10466469)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Keywords: 融合遺伝子 / 肉腫 / 機能解析

Outline of Annual Research Achievements

約100種類の肉腫関連融合遺伝子のクローニングを行い, マウス線維芽細胞である3T3細胞に導入し, focus formation assay及びmixed all nominated mutants in one method (MANO法)でのcell growth competition assayを行い, がん化能を評価した.
また, C2C12マウス筋芽細胞及びマウス間葉系幹細胞に遺伝子導入し, 分化誘導培地やバイオマテリアルを選択し分化誘導を行い, MANO法での細胞数変化を定量し, 融合遺伝子の分化抑制機能を評価すると共に, 評価に適した培養系を確立した.
さらに, トランスフォームしたがん細胞のみが生存可能なバイオマテリアル探索を行った. がん化を確認できた融合遺伝子を3T3細胞, C2C12細胞に導入したisogenicモデルで, 異なる蛍光蛋白質でラベルした親株とがん化株をポリマースライド上に播種し, 比較セルアレイを実施した. がん化株のみが有意に増殖・接着する合成ポリマーを蛍光イメージングにより抽出した.
3T3細胞, C2C12細胞, マウス間葉系幹細胞に肉腫関連融合遺伝子を導入したisogenic細胞株のRNA-seqを行い, 各融合遺伝子が発現制御する特異的な遺伝子発現パターンを比較し, 融合遺伝子のクラスタリング解析を行った. さらに, Gene Set Enrichment Analysis (GSEA)により各融合遺伝子の特徴となる発現遺伝子群やシグナルパスウェイを同定しクラスターごとに, 候補分子標的薬の選定を行った. MANO法を用いて候補薬剤評価実験を行い, 融合遺伝子特異的に作用する分子標的薬を同定した.

Research Progress Status

令和2年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

令和2年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Rapid reprogramming of tumour cells into cancer stem cells on double-network hydrogels (2021)
  • Author(s): Suzuka J, Tsuda M, Wang L, Kohsaka S, Kishida K, Semba S, Sugino H, Aburatani S, Frauenlob M, Kurokawa T, Kojima S, Ueno T, Ohmiya Y, Mano H, Yasuda K, Gong JP, Tanaka S
  • Journal Title: Nat Biomed Eng Volume: － Pages: －
  • DOI: 10.1038/s41551-021-00692-2.
  • Peer Reviewed
• [Journal Article] The initial assessment of expert panel performance in core hospitals for cancer genomic medicine in Japan (2021)
  • Author(s): Sunami K, Naito Y, Aimono E, Amano T, Ennishi D, Kage H, Kanai M, Komine K, Koyama T, Maeda T, Morita S, Sakai D, Kohsaka S, Tsuchihara K, Yoshino T
  • Journal Title: Int J Clin Oncol Volume: 26 Pages: 443-449
  • DOI: 10.1007/s10147-020-01844-1
  • Peer Reviewed / Open Access
• [Journal Article] Development of an optimal protocol for molecular profiling of tumor cells in pleural effusions at single-cell level (2021)
  • Author(s): Takeda Nakamura I, Ikegami M, Hasegawa N, Hayashi T, Ueno T, Kawazu M, Yagishita S, Goto Y, Shinno Y, Kojima Y, Takamochi K, Takahashi F, Takahashi K, Mano H, Kohsaka S
  • Journal Title: Cancer Sci Volume: － Pages: －
  • DOI: 10.1111/cas.14821.
  • Peer Reviewed / Open Access
• [Journal Article] Comprehensive molecular and clinicopathological profiling of desmoid tumors (2021)
  • Author(s): Kohsaka S*, Hirata M, Ikegami M, Ueno T, Kojima S, Sakai T, Ito K, Naka N, Ogura K, Kawai A, Iwata S, Okuma T, Yonemoto T, Kobayashi H, Suehara Y, Hiraga H, Kawamoto T, Motoi T, Oda Y, Matsubara D, Matsuda K, Nishida Y, and Mano H
  • Journal Title: European Journal of Cancer Volume: 145 Pages: 109-120
  • DOI: 10.1016/j.ejca.2020.12.001
  • Peer Reviewed / Open Access
• [Journal Article] Extensive functional evaluation of exon 20 insertion mutations of EGFR (2021)
  • Author(s): Hirose T, Ikegami M, Endo M, Matsumoto Y, Nakashima Y, Mano H, and Kohsaka S
  • Journal Title: Lung Cancer Volume: 152 Pages: 135-142
  • DOI: 10.1016/j.lungcan.2020.12.023
  • Peer Reviewed / Open Access
• [Journal Article] RET inhibition in novel patient-derived models of RET-fusion positive lung adenocarcinoma reveals a role for MYC upregulation (2020)
  • Author(s): Hayashi T, Odintsov I, Smith RS, Ishizawa K, Liu AJW, Delasos L, Kurzatkowski C, Tai H, Gladstone E, Vojnic M, Kohsaka S, Suzawa K, Liu Z, Kunte S, Mattar MS, Khodos I, Davare MA, Drilon A, Cheng E, Stanchina E, Ladanyi M, Somwar R
  • Journal Title: Dis Model Mech Volume: － Pages: －
  • DOI: 10.1242/dmm.047779.
  • Peer Reviewed / Open Access
• [Journal Article] Histological Characteristics of Lung Adenocarcinoma with Uncommon Actionable Alterations: Special Emphasis on MET exon 14 Skipping Alterations (2020)
  • Author(s): Hayashi T, Kohsaka S, Takamochi K, Kishikawa S, Ikarashi D, Sano K, Hara K, Onagi H, Suehara Y, Takahashi F, Saito T, Nakatsura T, Kitano S, Suzuki K, Yao T
  • Journal Title: Histopathology Volume: － Pages: －
  • DOI: 10.1111/his.14311.
  • Peer Reviewed
• [Journal Article] Genetic Testing Distinguishes Multiple Chondroid Chordomas with Neuraxial Bone Metastases from Multicentric Tumors (2020)
  • Author(s): Kobayashi H, Shin M, Makise N, Shinozaki-Ushiku A, Ikegami M, Taniguchi Y, Shinoda Y, Kohsaka S, Ushiku T, Oda K, Miyagawa K, Aburatani H, Mano H, Tanaka S
  • Journal Title: Case Rep Genet Volume: － Pages: －
  • DOI: 10.1155/2020/8877722.
  • Peer Reviewed / Open Access
• [Journal Article] Clinical practice guidance for next-generation sequencing in cancer diagnosis and treatment (edition 2) (2020)
  • Author(s): Naito Y, Aburatani H, Amano T, Baba E, Furukawa T, Hayashida T, Hiyama E, Ikeda S, Kanai M, Kato M, Kinoshita I, Kiyota N, Kohno T, Kohsaka S, et al.
  • Journal Title: Int J Clin Oncol Volume: － Pages: －
  • DOI: 10.1007/s10147-020-01831-6.
  • Peer Reviewed / Open Access
• [Journal Article] Identification of a Novel MAN1A1-ROS1 Fusion Gene Through mRNA-based Screening for Tyrosine Kinase Gene Aberrations in a Patient with Leiomyosarcoma (2020)
  • Author(s): Suehara Y, Kohsaka S, Hayashi T, Akaike K, Kurisaki-Arakawa A, Sato S, Kobayashi E, Mizuno S, Ueno T, Morii T, Okuma T, Kurihara T, Hasegawa N, Sano K, Sasa K, Okubo T, Kim Y, Mano H, Saito T
  • Journal Title: Clin Orthop Relat Res Volume: － Pages: －
  • DOI: 10.1097/CORR.0000000000001548.
  • Peer Reviewed
• [Journal Article] Nanostring-based screening for tyrosine kinase fusions in inflammatory myofibroblastic tumors (2020)
  • Author(s): Kurihara T, Suehara Y, Akaike K, Hayashi T, Kohsaka S, Ueno T, Hasegawa N, Takagi T, Sasa K, Okubo T, Kim Y, Mano H, Yao T, Kaneko K, Saito T
  • Journal Title: Sci Rep Volume: 10 Pages: 18724
  • DOI: 10.1038/s41598-020-75596-3.
  • Peer Reviewed / Open Access
• [Journal Article] Diffuse expression of MUC6 defines a distinct clinicopathological subset of pulmonary invasive mucinous adenocarcinoma (2020)
  • Author(s): Kishikawa S, Hayashi T, Saito T, Takamochi K, Kohsaka S, Sano K, Sasahara N, Sasa K, Kurihara T, Hara K, Suehara Y, Takahashi F, Suzuki K, Yao T
  • Journal Title: Mod Pathol Volume: － Pages: －
  • DOI: 10.1038/s41379-020-00690-w.
  • Peer Reviewed / Open Access
• [Journal Article] The Brain Microenvironment Induces DNMT1 Suppression and Indolence of Metastatic Cancer Cells (2020)
  • Author(s): Hirata E, Ishibashi K, Kohsaka S, Shinjo K, Kojima S, Kondo Y, Mano H, Yano S, Kiyokawa E, Sahai E
  • Journal Title: iScience Volume: 23 Pages: 101480
  • DOI: 10.1016/j.isci.2020.101480.
  • Peer Reviewed / Open Access
• [Journal Article] Clinicopathological characteristics of lung adenocarcinoma with compound EGFR mutations (2020)
  • Author(s): Hayashi T, Kohsaka S, Takamochi K, Hara K, Kishikawa S, Sano K, Takahashi F, Suehara Y, Saito T, Takahashi K, Suzuki K, Yao T
  • Journal Title: Hum Pathol Volume: 103 Pages: 42-51
  • DOI: 10.1016/j.humpath.2020.07.007
  • Peer Reviewed
• [Journal Article] Comprehensive clinicopathological and molecular analysis of primary malignant melanoma of the oesophagus (2020)
  • Author(s): Tsuyama S, Kohsaka S, Hayashi T, Suehara Y, Hashimoto T, Kajiyama Y, Tsurumaru M, Ueno T, Mano H, Yao T, Saito T
  • Journal Title: Histopathology Volume: － Pages: －
  • DOI: 10.1111/his.14210.
  • Peer Reviewed
• [Journal Article] Genomic Profiling of Multiple Primary Cancers Including Synchronous Lung Adenocarcinoma and Bilateral Malignant Mesotheliomas: Identification of a Novel BAP1 Germline Variant (2020)
  • Author(s): Shinozaki-Ushiku A, Kohsaka S, Kage H, Oda K, Miyagawa K, Nakajima J, Aburatani H, Mano H, Ushiku T
  • Journal Title: Pathology International Volume: － Pages: －
  • DOI: 10.1111/pin.12977.
  • Peer Reviewed
• [Journal Article] Assessment of Predictive Biomarkers of the Response to Pazopanib Based on an Integrative Analysis of High-grade Soft-tissue Sarcomas: Analysis of a Tumor Sample From a Responder and Patients With Other Soft-tissue Sarcomas (2020)
  • Author(s): Suehara Y, Kohsaka S, Yamaguchi S, Hayashi T, Kurihara T, Sano K, Sasa K, Akaike K, Ueno T, Kojima S, Ikegami M, Mizuno S, Okubo T, Kim Y, Kaneko K, Saito T, Kato S, Mano H
  • Journal Title: Clin Orthop Relat Res Volume: － Pages: －
  • DOI: 10.1097/CORR.0000000000001322.
  • Peer Reviewed
• [Journal Article] High-Throughput Functional Evaluation of BRCA2 Variants of Unknown Significance (2020)
  • Author(s): Ikegami M, Kohsaka S*, Ueno T, Momozawa Y, Tamura K, Shimomura A, Hosoya N, Kobayashi H, Tanaka S, Mano H
  • Journal Title: Nature Communications Volume: 11 Pages: 2573
  • DOI: 10.1038/s41467-020-16141-8
  • Peer Reviewed / Open Access
• [Journal Article] がんゲノム医療におけるVUS（variant of unknown significance）の評価 (2020)
  • Author(s): 高阪 真路
  • Journal Title: がん分子標的治療 Volume: 18 Pages: 72-77
• [Journal Article] がんゲノム医療の推進に向けた知識データベース構築と維持 (2020)
  • Author(s): 高阪 真路, 小山 隆文, 角南 久仁子, 下井 辰徳, 柴田 大朗
  • Journal Title: 保健医療科学 Volume: 69 Pages: 260-273
  • DOI: 10.20683/jniph.69.3_260
  • Open Access
• [Presentation] 発がん関連遺伝子における生殖細胞系列バリアントのハイスループット機能解析法 (2021)
  • Author(s): 高阪真路
  • Organizer: 遺伝性腫瘍セミナー
  • Invited
• [Presentation] Liquid型パネルとC-CATリポジトリの活用 (2021)
  • Author(s): 高阪真路
  • Organizer: 第18回日本臨床腫瘍学会学術集会
  • Invited
• [Presentation] がん個別化医療の実現に向けた戦略的研究開発 (2021)
  • Author(s): 高阪真路
  • Organizer: AI技術が切り拓く新たなリウマチ学に向けた教育シンポジウム
  • Invited
• [Presentation] がんゲノム医療の現況と展望 (2020)
  • Author(s): 高阪真路
  • Organizer: 第33回横浜市立大学がんプロ合同セミナー
  • Invited
• [Presentation] 個別化がんゲノム医療の実現に向けた知識データベースの構築 (2020)
  • Author(s): 高阪真路
  • Organizer: 第58回日本癌治療学会学術集会
  • Invited
• [Presentation] がん個別化医療の実現に向けた戦略的研究開発 (2020)
  • Author(s): 高阪真路
  • Organizer: AI技術が切り拓く新たなリウマチ学に向けた教育シンポジウム
  • Invited
• [Presentation] がんゲノム医療におけるリキッドバイオプシーの実践 (2020)
  • Author(s): 高阪真路
  • Organizer: 第109回日本病理学会総会
  • Invited
• [Book] C-CAT調査結果の読み方. がんゲノム医療遺伝子パネル検査実践ガイド (2020)
  • Author(s): 高阪 真路
  • Total Pages: 252
  • Publisher: 医学書院
  • ISBN: 978-4-260-04246-8