分子標的治療耐性化メカニズムの網羅的探索

2021 Fiscal Year Annual Research Report

• Project/Area Number: 21H02795
• 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, 分野長 (00627119)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Keywords: ゲノム医学 / 薬剤耐性 / がん遺伝子 / ハイスループット機能解析

Outline of Annual Research Achievements

1. がん遺伝子の網羅的機能解析
68種類の遺伝子、243バリアントについて、3T3マウス線維芽細胞株にがん遺伝子を導入し、3T3 focus formation assay及びMANO法でのcell competition assayによるがん化能評価を行った。 さらに、肺がん細胞株、乳がん細胞株、メラノーマ細胞株に遺伝子を導入し、表現型の変化をfocus formation assayおよびsoft agar colony formation assayで評価した。
2. MANO法を用いた分子標的薬耐性化メカニズムの解明
特定の遺伝子経路に依存し、依存したシグナル伝達経路を阻害する分子標的薬に対しての感受性を示す細胞株を準備した。PC9 EGFR変異陽性肺がん細胞株、H2228 EML4-ALK fusion陽性肺がん細胞株、MDA-MB-361 PIK3CA変異陽性乳がん細胞株、A375 BRAF変異陽性メラノーマ細胞株にがん遺伝子243バリアントを導入して、各分子標的薬に対する薬剤感受性実験をMANO法を用いて行った。EGFR、ALK、BRAF阻害薬に対する耐性マーカーが複数同定された。さらに、取得された分子標的薬耐性バリアントを個別に細胞株に導入し、薬剤耐性化を検証した。耐性バリアントを導入した細胞と親株とを薬剤投与下でmRNAの発現解析、タンパク質リン酸化の解析を行い、耐性化のメカニズムを評価した。

Current Status of Research Progress

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

Reason

予定していた、がん遺伝子の網羅的機能評価および分子標的薬耐性化スクリーニングを行い、いくつかの新規のバイオマーカーを同定することができた。

Strategy for Future Research Activity

スクリーニングにて候補として同定された薬剤耐性マーカーについて、個別の細胞実験による検証を行うと共に、複数の耐性バリアントのメカニズムを比較することで、耐性化の網羅的評価を行う。

Research Products

• [Journal Article] Genetic diagnosis of pseudomyxoma peritonei originating from mucinous borderline tumor inside an ovarian teratoma (2022)
  • Author(s): Taguchi A, Rokutan H, Oda K, Tanikawa M, Tanimoto S, Sone K, Mori M, Tsuruga T, Kohsaka S, Tatsuno K, Shinozaki-Ushiku A, Miyagawa K, Mano H, Aburatani H, Ushiku T, Osuga Y.
  • Journal Title: BMC Med Genomics Volume: 15 Pages: 51
  • DOI: 10.1186/s12920-022-01188-x
  • Peer Reviewed / Open Access
• [Journal Article] Composite Clonal Analysis Reveals Transition of NSCLC Subtypes Through Accumulation of Gene Mutations: A Case Report (2022)
  • Author(s): Ando T, Kage H, Shinozaki-Ushiku A, Tatsuno K, Tsutsumi S, Nagayama K, Nakajima J, Kohsaka S, Miyagawa K, Aburatani H, Mano H, Nagase T.
  • Journal Title: JTO Clin Res Rep Volume: 3 Pages: 100277
  • DOI: 10.1016/j.jtocrr.2022.100277
  • Peer Reviewed / Open Access
• [Journal Article] The Role of the EZH2 and H3K27me3 Expression as a Predictor of Clinical Outcomes in Salivary Duct Carcinoma Patients: A Large-Series Study With Emphasis on the Relevance to the Combined Androgen Blockade and HER2-Targeted Therapy (2022)
  • Author(s): Saigusa N, Hirai H, Kohsaka S, et al.
  • Journal Title: Front Oncol Volume: 11 Pages: 779882
  • DOI: 10.3389/fonc.2021.779882
  • Peer Reviewed
• [Journal Article] CIC-Mediated Modulation of MAPK Signaling Opposes Receptor Tyrosine Kinase Inhibitor Response in Kinase-Addicted Sarcoma (2022)
  • Author(s): Odintsov I, Ortiz MV, Khodos I, Mattar MS, Lui AJW, Kohsaka S, de Stanchina E, Bender JLG, Ladanyi M, Somwar R.
  • Journal Title: Cancer Res Volume: 82 Pages: 1110-1127
  • DOI: 10.1158/0008-5472.CAN-21-1397
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Comparative Study on the Efficacy and Exposure of Molecular Target Agents in Non-small Cell Lung Cancer PDX Models with Driver Genetic Alterations (2022)
  • Author(s): Jo H, Yagishita S, Hayashi Y, Ryu S, Suzuki M, Kohsaka S, Ueno T, Matsumoto Y, Horinouchi H, Ohe Y, Watanabe SI, Motoi N, Yatabe Y, Mano H, Takahashi K, Hamada A.
  • Journal Title: Mol Cancer Ther Volume: 21 Pages: 359-370
  • DOI: 10.1158/1535-7163.MCT-21-0371
  • Peer Reviewed
• [Journal Article] Distinct properties of pure- and mixed-type high-grade fetal lung adenocarcinomas by genetic profiling and transcription factor expression (2022)
  • Author(s): Kishikawa S, Hayashi T, Saito T, Takamochi K, Sasa K, Suehara Y, Takahashi F, Sasahara N, Kohsaka S, Suzuki K, Yao T.
  • Journal Title: Virchows Arch Volume: 480 Pages: 609-619
  • DOI: 10.1007/s00428-021-03247-7
  • Peer Reviewed
• [Journal Article] Preclinical assessment of combination therapy of EGFR tyrosine kinase inhibitors in a highly heterogeneous tumor model (2022)
  • Author(s): Ikeuchi H, Hirose T, Ikegami M, Takamochi K, Suzuki K, Mano H, Kohsaka S.
  • Journal Title: Oncogene Volume: 41 Pages: 2470-2479
  • DOI: 10.1038/s41388-022-02263-4
  • Peer Reviewed / Open Access
• [Journal Article] MicroSEC filters sequence errors for formalin-fixed and paraffin-embedded samples (2021)
  • Author(s): Ikegami M, Kohsaka S, Hirose T, Ueno T, Inoue S, Kanomata N, Yamauchi H, Mori T, Sekine S, Inamoto Y, Yatabe Y, Kobayashi H, Tanaka S, Mano H.
  • Journal Title: Commun Biol Volume: 4 Pages: 1396
  • DOI: 10.1038/s42003-021-02930-4
  • Peer Reviewed / Open Access
• [Journal Article] Scapular Angiomatoid Fibrous Histiocytoma with EWSR1-CREB1 Fusion in an Adult Patient (2021)
  • Author(s): Kobayashi H, Makise N, Shinozaki-Ushiku A, Ishibashi Y, Ikegami M, Kohsaka S, Ushiku T, Oda K, Miyagawa K, Aburatani H, Mano H, Tanaka S.
  • Journal Title: Case Rep Orthop Volume: 2021 Pages: 9434222
  • DOI: 10.1155/2021/9434222
  • Peer Reviewed / Open Access
• [Journal Article] Programmed death-ligand 1 expression and its associations with clinicopathological features, prognosis, and driver oncogene alterations in surgically resected lung adenocarcinoma (2021)
  • Author(s): Takamochi K, Hara K, Hayashi T, Kohsaka S, Takahashi F, Suehara Y, Suzuki K.
  • Journal Title: Lung Cancer Volume: 161 Pages: 163-170
  • DOI: 10.1016/j.lungcan.2021.09.011
  • Peer Reviewed
• [Journal Article] Highly sensitive fusion detection using plasma cell-free RNA in non-small-cell lung cancers (2021)
  • Author(s): Hasegawa N, Kohsaka S, Kurokawa K, Shinno Y, Takeda Nakamura I, Ueno T, Kojima S, Kawazu M, Suehara Y, Ishijima M, Goto Y, Kojima Y, Yonemori K, Hayashi T, Saito T, Shukuya T, Takahashi F, Takahashi K, Mano H.
  • Journal Title: Cancer Sci Volume: 112 Pages: 4393-4403
  • DOI: 10.1111/cas.15084
  • Peer Reviewed / Open Access
• [Journal Article] Comprehensive functional evaluation of variants of fibroblast growth factor receptor genes in cancer (2021)
  • Author(s): Nakamura IT, Kohsaka S, Ikegami M, Ikeuchi H, Ueno T, Li K, Beyett TS, Koyama T, Shimizu T, Yamamoto N, Takahashi F, Takahashi K, Eck MJ, Mano H.
  • Journal Title: NPJ Precis Oncol Volume: 5 Pages: 66
  • DOI: 10.1038/s41698-021-00204-0
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Comprehensive molecular profiling of pulmonary pleomorphic carcinoma (2021)
  • Author(s): Nagano M, Kohsaka S, Hayashi T, Ueno T, Kojima S, Shinozaki-Ushiku A, Morita S, Tsuda M, Tanaka S, Shinohara T, Omori Y, Sugaya F, Kato H, Narita Y, Nakajima J, Suzuki K, Takamochi K, Mano H.
  • Journal Title: NPJ Precis Oncol Volume: 5 Pages: 57
  • DOI: 10.1038/s41698-021-00201-3
  • Peer Reviewed / Open Access
• [Journal Article] Appropriate use of cancer comprehensive genome profiling assay using circulating tumor DNA (2021)
  • Author(s): Sunami K, Bando H, Yatabe Y, Naito Y, Takahashi H, Tsuchihara K, Toyooka S, Mimori K, Kohsaka S, Uetake H, Kinoshita I, Komine K, Takeda M, Hayashida T, Tamura K, Nishio K, Yamamoto N; Working Group of a Joint Task Force of Three Academic Societies for the Promotion of Cancer Genomic Medicine.
  • Journal Title: Cancer Sci Volume: 112 Pages: 3911-3917
  • DOI: 10.1111/cas.15022
  • Peer Reviewed / Open Access
• [Journal Article] Transcription start site-level expression of thyroid transcription factor 1 isoforms in lung adenocarcinoma and its clinicopathological significance (2021)
  • Author(s): Sano K, Hayashi T, Suehara Y, Hosoya M, Takamochi K, Kohsaka S, Kishikawa S, Kishi M, Saito S, Takahashi F, Kaneko K, Suzuki K, Yao T, Ishijima M, Saito T.
  • Journal Title: J Pathol Clin Res Volume: 7 Pages: 361-374
  • DOI: 10.1002/cjp2.213
  • Peer Reviewed / Open Access
• [Journal Article] Correction to: 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: 1007
  • DOI: 10.1007/s10147-021-01897-w
  • Peer Reviewed
• [Journal Article] Development of an optimal protocol for molecular profiling of tumor cells in pleural effusions at single-cell level (2021)
  • Author(s): Nakamura IT, 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: 112 Pages: 2006-2019
  • DOI: 10.1111/cas.14821
  • Peer Reviewed / Open Access
• [Presentation] がんゲノム医療におけるリキッドバイオプシーの有用性と現状 (2021)
  • Author(s): 高阪真路
  • Organizer: 第80日本癌学会学術総会
• [Presentation] High throughput functional analysis for the clinical annotation in cancer genome medicine (2021)
  • Author(s): Shinji Kohsaka
  • Organizer: 1stJCA-AACR Precision Cancer Medicine International Conference
  • Int'l Joint Research / Invited
• [Presentation] がんゲノム医療におけるリキッドバイオプシーの有用性と現状 (2021)
  • Author(s): 高阪真路
  • Organizer: 第62回日本臨床細胞学会総会春期大会
  • Int'l Joint Research / Invited
• [Book] がんゲノム病理学 (2021)
  • Author(s): 高阪真路
  • Total Pages: 234
  • Publisher: 文光堂
  • ISBN: 978-4-8306-0485-0