Development of a diagnostic platform for precision oncology by applying reverse-phase phosphoprotein array to an analysis of formalin-fixed paraffin-embedded (FFPE) tissue

• Project/Area Number: 20K07690
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 50020:Tumor diagnostics and therapeutics-related
• Research Institution: National Cancer Center Japan
• Principal Investigator: MASUDA MARI 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 (70435717)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2022: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2021: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2020: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
• Keywords: RPPA / Wnt シグナル / シグナル伝達 / リン酸化タンパク質 / バイオマーカー / 治療標的 / トレンスレーショナル・リサーチ / プロテオミクス / がん高精度医療 / 薬剤抵抗性 / 分子標的薬 / シグナル経路 / タンパク質リン酸化 / FFPE / 薬剤耐性 / 逆相タンパクアレイ / ホルマリン固定・パラフィン包埋組織（FFPE） / 高精度医療 / コンパニオンマーカー / Precision Oncology

Outline of Research at the Start

がんゲノム高精度医療の改善に向けて、多くの分子標的治療薬の標的となるタンパク及び下流のタンパクの活性化を逆相リン酸化タンパクアレイ（RPPA）法により把握できれば、分子標的治療薬による適切な治療戦略の提示や治療後の獲得耐性の克服が可能となると期待される。本研究では、一般的に病理診断で用いられるホルマリン固定・パラフィン包埋組織（FFPE）を用いた逆相リン酸化タンパクアレイ（FFPE-RPPA）法の技術開発を第一目的とする。更に、本研究により確立される基盤を用いて、申請者らが先行研究で見出した中皮腫に対するFGFR阻害剤のコンパニオンマーカーの開発と検証を行うことを第二の目的とする。

Outline of Final Research Achievements

In this study, we aimed to optimize the detection of phosphoproteins in formalin-fixed and paraffin-embedded (FFPE) tissue, which is commonly used for pathological diagnosis, using a reverse-phase phosphoprotein array method. The FFPE-RPPA method is expected to reinforce highly accurate precision cancer medicine in the future. We first improved our original RPPA technology by optimizing the protein concentration and the signal detection method suitable for the FFPE-RPPA method. With this refined FFPE-RPPA platform, we next compared the expression of phosphoproteins in the protein samples extracted from either the cells or the FFPE block of EGFR inhibitor-treated (EGFRi-)sensitive and resistant non-small cell cancer cells. As a result, greater inhibitor-induced signal variation was detected with the optimized FFPE-RPPA method.

Academic Significance and Societal Importance of the Research Achievements

個々のがん患者に適切な薬剤を見出すがんゲノム医療が一般化しつつあるが、薬剤抵抗性や臨床試験登録の地域格差も含め克服すべき問題は多く存在する。殆どの分子標的治療薬の標的となるキナーゼや下流シグナルのタンパク質の活性化（リン酸化）を、一般的に病理診断材料であるホルマリン固定・パラフィン包埋組織（FFPE）を用いて逆相リン酸化タンパクアレイ（RPPA）法で正確に把握できれば、多剤併用を含む適切な治療戦略の提示や薬剤に対する初回耐性及び治療後の獲得耐性の克服に貢献できることが期待され社会的意義は高い。また得られる知見は今後の薬剤開発への重要な知見となり学術的重要性も高い。

Research Products

• [Journal Article] Stromal reprogramming through dual PDGFRα/β blockade boosts the efficacy of anti-PD-1 immunotherapy in fibrotic tumors. (2022)
  • Author(s): T. Akiyama, T. Yasuda, T. Uchihara, N. Yasuda-Yoshihara, BJY. Tan, A. Yonemura, T. Semba, J. Yamasaki, Y. Komohara, K. Ohnishi, F. Wei, L. Fu, J. Zhang, F. Kitamura, K. Yamashita, K. Eto, S. Iwagami, H. Tsukamoto, T. Umemoto, M. Masuda, O. Nagano, Y. Satou, H. Saya, P. Tan, H. Baba, T. Ishimoto
  • Journal Title: Cancer Res. Volume: 83 Issue: 5 Pages: 753-770
  • DOI: 10.1158/0008-5472.can-22-1890
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Potential utility of reverse-phase protein array for refining precision cancer medicine (2022)
  • Author(s): Mari Masuda
  • Journal Title: Electrophoresis Letters Volume: 66 Issue: 1 Pages: 31-34
  • DOI: 10.2198/electroph.66.31
  • ISSN: 2189-2628, 2189-2636
  • Peer Reviewed
• [Journal Article] Clinical and Tumor Characteristics of Patients with High Serum Levels of Growth Differentiation Factor 15 in Advanced Pancreatic Cancer. (2021)
  • Author(s): Suzuki H, Mitsunaga S, Ikeda M, Aoyama T, Yoshizawa K, Yoshimatsu H, Kawai N, Masuda M, Miura T, Ochiai A.
  • Journal Title: Cancers Volume: 13 Issue: 19 Pages: 4842-4842
  • DOI: 10.3390/cancers13194842
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Early dynamics of circulating tumor DNA predict chemotherapy responses for patients with esophageal cancer (2021)
  • Author(s): Ryosuke Fujisawa , Takeshi Iwaya, Fumitaka Endo, Masashi Idogawa, Noriyuki Sasaki, Hayato Hiraki, Shoichiro Tange, Tomomi Hirano, Yuka Koizumi , Masakazu Abe, Tomoko Takahashi, Mizunori Yaegashi , Yuji Akiyama, Mari Masuda, Akira Sasaki, Fumiaki Takahashi, Yasushi Sasaki, Takashi Tokino, Satoshi S Nishizuka
  • Journal Title: Carcinogenesis Volume: 42(10) Issue: 10 Pages: 1239-1249
  • DOI: 10.1093/carcin/bgab088
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Direct conversion of osteosarcoma to adipocytes by targeting TNIK (2021)
  • Author(s): Hirozane Toru、Masuda Mari、Sugano Teppei、Sekita Tetsuya、Goto Naoko、Aoyama Toru、Sakagami Takato、Uno Yuko、Moriyama Hideki、Sawa Masaaki、Asano Naofumi、Nakamura Masaya、Matsumoto Morio、Nakayama Robert、Kondo Tadashi、Kawai Akira、Kobayashi Eisuke、Yamada Tesshi
  • Journal Title: JCI Insight Volume: 6 Issue: 3
  • DOI: 10.1172/jci.insight.137245
  • Peer Reviewed / Open Access
• [Journal Article] Pharmacological blockage of transforming growth factor-β signalling by a Traf2- and Nck-interacting kinase inhibitor, NCB-0846 (2020)
  • Author(s): Sugano Teppei、Masuda Mari、Takeshita Fumitaka、Motoi Noriko、Hirozane Toru、Goto Naoko、Kashimoto Shigeki、Uno Yuko、Moriyama Hideki、Sawa Masaaki、Nagakawa Yuichi、Tsuchida Akihiko、Seike Masahiro、Gemma Akihiko、Yamada Tesshi
  • Journal Title: British Journal of Cancer Volume: 124 Issue: 1 Pages: 228-236
  • DOI: 10.1038/s41416-020-01162-3
  • Peer Reviewed / Open Access
• [Journal Article] Extracellular Vesicles from Cancer-Associated Fibroblasts Containing Annexin A6 Induces FAK-YAP Activation by Stabilizing β1 Integrin, Enhancing Drug Resistance (2020)
  • Author(s): Uchihara T, Miyake K, Yonemura A, Komohara Y, Itoyama R, Koiwa M, Yasuda T, Arima K, Harada K, Eto K, Hayashi H, Iwatsuki M, Iwagami S, Baba Y, Yoshida N, Yashiro M, Masuda M, Ajani Jaffer A, Tan Patrick, Baba H, Ishimoto T
  • Journal Title: Cancer Research Volume: 80 Issue: 16 Pages: 3222-3235
  • DOI: 10.1158/0008-5472.can-19-3803
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Feasibility of Targeting Traf2-and-Nck-Interacting Kinase in Synovial Sarcoma (2020)
  • Author(s): Sekita Tetsuya、Yamada Tesshi、Kobayashi Eisuke、Yoshida Akihiko、Hirozane Toru、Kawai Akira、Uno Yuko、Moriyama Hideki、Sawa Masaaki、Nagakawa Yuichi、Tsuchida Akihiko、Matsumoto Morio、Nakamura Masaya、Nakayama Robert、Masuda Mari
  • Journal Title: Cancers Volume: 12 Issue: 5 Pages: 1258-1258
  • DOI: 10.3390/cancers12051258
  • Peer Reviewed / Open Access
• [Journal Article] Whole-exome and RNA sequencing of pulmonary carcinoid reveals chromosomal rearrangements associated with recurrence (2020)
  • Author(s): Miyanaga Akihiko、Masuda Mari、Motoi Noriko、Tsuta Koji、Nakamura Yuka、Nishijima Nobuhiko、Watanabe Shun-ichi、Asamura Hisao、Tsuchida Akihiko、Seike Masahiro、Gemma Akihiko、Yamada Tesshi
  • Journal Title: Lung Cancer Volume: 145 Pages: 85-94
  • DOI: 10.1016/j.lungcan.2020.03.027
  • Peer Reviewed
• [Journal Article] Prognostic impact of ACTN4 gene copy number alteration in hormone receptor-positive, HER2-negative, node-negative invasive breast carcinoma (2020)
  • Author(s): Sugano T, Yoshida M, Masuda M, Ono M, Tamura K, Kinoshita T, Tsuda H, Honda K, Gemma A, Yamada T
  • Journal Title: Br J Cancer. Volume: 122 Issue: 12 Pages: 1811-1817
  • DOI: 10.1038/s41416-020-0821-y
  • Peer Reviewed
• [Presentation] がん高精度医療の更なる改善に向けた逆相タンパクアレイ技術基盤の有用性 (2022)
  • Author(s): 増田万里、中川莉杏、近藤格
  • Organizer: 第72回日本プロテオーム学会2022年大会・ＪＨＵＰＯ第20回大会・シンポジウム
  • Invited
• [Presentation] がん医療へ応用可能な逆相タンパクアレイ（Reverse-phase protein array; RPPA）技術基盤の最適化 (2021)
  • Author(s): 中川莉杏、増田万里
  • Organizer: 第44回日本分子生物学会年会
• [Presentation] 逆相タンパクアレイ(Reverse-Phase Protein Array;RPPA)技術基盤の最適化 (2021)
  • Author(s): 中川莉杏、増田万里
  • Organizer: 第72回日本電気泳動学会総会
• [Presentation] 逆相タンパクアレイ基盤のがん高精度医療への応用 (2021)
  • Author(s): 増田万里
  • Organizer: 第72回日本電気泳動学会総会
  • Invited
• [Presentation] Utility of Reverse-Phase Protein Array for Refining Precision Oncology (2020)
  • Author(s): Mari Masuda
  • Organizer: Korean Society for Biochemistry and Molecular Biology （KSBMB） International Conference 2020
  • Int'l Joint Research / Invited
• [Presentation] Identification of therapeutic target and potential companion marker for molecular-targeted therapy of malignant pleural mesothelioma (2020)
  • Author(s): Toru Aoyama, Noriko Motoi, Naoko Goto, Riko Nakagaw, Masahiro Seike, Mari Masuda
  • Organizer: 第７９回日本癌学会学術総会
  • Int'l Joint Research
• [Remarks] 患者個人に合わせた超高感度血液検査で食道がん再発を早期に検出する方法を開発 (プレスリリース；PR）
  • URL: https://www.ncc.go.jp/jp/information/pr_release/2020/1012/index.html
• [Remarks] 骨肉腫を脂肪細胞へ変化させることに成功、TNIK阻害薬活用による新規治療法開発への期待 (PR)）
  • URL: https://www.ncc.go.jp/jp/information/pr_release/2021/0205/index.html