Discovery of novel cancer treatment with CRIPR/Cas9 screening

• Project/Area Number: 18K08184
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 53030:Respiratory medicine-related
• Research Institution: Keio University (2019-2020); Kitasato University (2018)
• Principal Investigator: TERAI HIDEKI 慶應義塾大学, 医学部(信濃町), 特任助教 (50445293)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2020: ¥390,000 (Direct Cost: ¥300,000、Indirect Cost: ¥90,000); Fiscal Year 2019: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000); Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: EGFR-TKI / CRISPR/Cas9 / SHOC2 / MAPK / EGFR / lung cancer / celastrol / drug persister / 非小細胞肺癌 / CRISPR screening / drug resistance

Outline of Final Research Achievements

We utilized CRISPR/Cas9 screening in EGFR mutation-positive non-small cell lung cancer cell lines to identify genes involved in EGFR-TKI resistance.
In addition to previously reported genes like AXL and FGFR1, we identified SHOC2, a scaffold protein coding gene involved in the activity of the MAP kinase pathway, as a gene involved in drug resistance. In EGFR mutation-positive lung cancer cell lines, we found a correlation between the expression level of SHOC2 and EGFR-TKI sensitivity.
In addition, we found that SHOC2 expression was altered before and after EGFR-TKI treatment in clinical lung cancer specimens.

Academic Significance and Societal Importance of the Research Achievements

肺癌は日本人の癌死亡原因の中で臓器別第一位であり、予後不良の疾患である。分子標的薬の登場により、患者予後の改善を認めているものの、分子標的薬のみでの根治は困難である。そのため、薬剤耐性機構の解明とその克服は重要な課題であるが、本研究においては、日本人非小細胞肺癌で最も割合の高い遺伝子変異であるEGFR遺伝子変異陽性肺癌に対する特効薬であるEGFR-TKIに対する薬剤耐性に関わる新規遺伝子として、SHOC2を同定し、その薬剤感受性に影響を及ぼすメカニズムの一端を明らかにした。今後、SHOC2を標的とした創薬やバイオマーカーとして活用することで、肺癌薬剤耐性の克服につながることを期待する。

Research Products

• [Journal Article] Functional dissection of the KRAS G12C mutation by comparison among multiple oncogenic driver mutations in a lung cancer cell line model (2021)
  • Author(s): Kobayashi Keigo、Terai Hideki、Yasuda Hiroyuki、Hamamoto Junko、Hayashi Yuichiro、Takeuchi Osamu、Mitsuishi Yoichiro、Masuzawa Keita、Manabe Tadashi、Ikemura Shinnosuke、Kawada Ichiro、Suzuki Yukio、Soejima Kenzo、Fukunaga Koichi
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 534 Pages: 1-7
  • DOI: 10.1016/j.bbrc.2020.11.110
  • Peer Reviewed
• [Journal Article] Intracellular levels of reactive oxygen species correlate with ABT‐263 sensitivity in non‐small‐cell lung cancer cells (2020)
  • Author(s): Ohgino Keiko、Terai Hideki、Yasuda Hiroyuki、Nukaga Shigenari、Hamamoto Junko、Tani Tetsuo、Kuroda Aoi、Arai Daisuke、Ishioka Kota、Masuzawa Keita、Ikemura Shinnosuke、Kawada Ichiro、Naoki Katsuhiko、Fukunaga Koichi、Soejima Kenzo
  • Journal Title: Cancer Science Volume: 111 Issue: 10 Pages: 3793-3801
  • DOI: 10.1111/cas.14569
  • Peer Reviewed
• [Journal Article] SHOC2 Is a Critical Modulator of Sensitivity to EGFR-TKIs in Non-Small Cell Lung Cancer Cells (2020)
  • Author(s): Terai Hideki、Hamamoto Junko、Emoto Katsura、Masuda Takeshi、Manabe Tadashi、Kuronuma Satoshi、Kobayashi Keigo、Masuzawa Keita、Ikemura Shinnosuke、Nakayama Sohei、Kawada Ichiro、Suzuki Yusuke、Takeuchi Osamu、Suzuki Yukio、Ohtsuki Sumio、Yasuda Hiroyuki、Soejima Kenzo、Fukunaga Koichi
  • Journal Title: Molecular Cancer Research Volume: 19 Issue: 2 Pages: 317-328
  • DOI: 10.1158/1541-7786.mcr-20-0664
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] IGF2 Autocrine-Mediated IGF1R Activation Is a Clinically Relevant Mechanism of Osimertinib Resistance in Lung Cancer (2020)
  • Author(s): Manabe Tadashi、Yasuda Hiroyuki、Terai Hideki、Kagiwada Harumi、Hamamoto Junko、Ebisudani Toshiki、Kobayashi Keigo、Masuzawa Keita、Ikemura Shinnosuke、Kawada Ichiro、Hayashi Yuichiro、Fukui Kazuhiko、Horimoto Katsuhisa、Fukunaga Koichi、Soejima Kenzo
  • Journal Title: Molecular Cancer Research Volume: 印刷中 Issue: 4 Pages: 549-559
  • DOI: 10.1158/1541-7786.mcr-19-0956
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Monomer Preference of EGFR Tyrosine Kinase Inhibitors Influences the Synergistic Efficacy of Combination Therapy with Cetuximab. (2019)
  • Author(s): Oashi A, Yasuda H, Kobayashi K, Tani T, Hamamoto J, Masuzawa K, Manabe T, Terai H, Ikemura S, Kawada I, Naoki K, Soejima K
  • Journal Title: Mol Cancer Ther Volume: 18 Issue: 9 Pages: 1593-1601
  • DOI: 10.1158/1535-7163.mct-18-1036
  • Peer Reviewed
• [Journal Article] Efficacy of afatinib or osimertinib plus cetuximab combination therapy for non-small-cell lung cancer with EGFR exon 20 insertion mutations (2019)
  • Author(s): Hasegawa H, Yasuda H, Hamamoto J, Masuzawa K, Tani T, Nukaga S, Hirano T, Kobayashi K, Manabe T, Terai H, Ikemura S, Kawada I, Naoki K, Soejima K.
  • Journal Title: Lung Cancer Volume: 127 Pages: 146-52
  • DOI: 10.1016/j.lungcan.2018.11.039
  • Peer Reviewed
• [Presentation] EGFRチロシンキナーゼ阻害剤の初期耐性におけるSHOC2の役割 (2019)
  • Author(s): 浜本 純子、寺井 秀樹、眞鍋 維志、小林 慧悟、増澤 啓太、安田 浩之、川田 一郎、副島 研造
  • Organizer: 第78回日本癌学会学術総会