Systems analysis of intrinsic resistance to MEK inhibitor in KRas- or BRaf-mutataed cancer cells

• Project/Area Number: 26290053
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Tumor therapeutics
• Research Institution: Okazaki Research Facilities, National Institutes of Natural Sciences (2016); Kyoto University (2014-2015)
• Principal Investigator: AOKI Kazuhiro 大学共同利用機関法人自然科学研究機構(岡崎共通研究施設), 岡崎統合バイオサイエンスセンター, 教授 (80511427)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥17,290,000 (Direct Cost: ¥13,300,000、Indirect Cost: ¥3,990,000); Fiscal Year 2016: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000); Fiscal Year 2015: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000); Fiscal Year 2014: ¥10,920,000 (Direct Cost: ¥8,400,000、Indirect Cost: ¥2,520,000)
• Keywords: 内因的抵抗性 / FRETイメージング / 数理モデル / シミュレーション / KRas / BRaf / ERK / PI3K / FRET / Akt / イメージング / レポーター

Outline of Final Research Achievements

In this study, we analyzed molecular mechanisms of intrinsic resistance to MEK inhibitor in BRaf- or KRas-mutated cancer cells. In BRaf-mutated cancer cells, MEK inhibitor linearly reduced ERK activity and cell proliferation rate, whereas KRas-mutated cancer cells showed non-linear relation between ERK activity and cell proliferation induced by MEK inhibitor, thereby showing resistance to MEK inhibitor. This was mediated by mTOR pathway, and we confirmed that co-treatment with MEK inhibitor and PI3K/mTOR inhibitor synergistically decreased cell proliferation rate. The molecular mechanisms underlying mTOR-mediated intrinsic resistance to MEK inhibitor in KRas-mutant cancer cells were further analyzed by experiments and mathematical modeling. We found that not only feedback regulations between ERK and mTOR pathways but also transcriptional regulation by ERK pathway substantially contributed to the intrinsic resistance to MEK inhibitor.

Research Products

• [Journal Article] Multiplexed Fluorescence Imaging of ERK and Akt Activities and Cell-cycle Progression (2016)
  • Author(s): Maryu G, Matsuda M, Aoki K
  • Journal Title: Cell Structure and Function Volume: 41 Issue: 2 Pages: 81-92
  • DOI: 10.1247/csf.16007
  • NAID: 130005165901
  • ISSN: 0386-7196, 1347-3700
  • Peer Reviewed / Open Access
• [Journal Article] Synergistic antitumor effects of combination of PI3K/mTOR and MEK inhibition (SAR245409 and pimasertib) in mucinous ovarian carcinoma cells by fluorescence resonance energy transfer imaging. (2016)
  • Author(s): Inaba K, Oda K, Aoki K, Sone K, Ikeda Y, Miyasaka A, Kashiyama T, Fukuda T, Makii C, Arimoto T, Wada-Hiraike O, Kawana K, Yano T, Osuga Y, Fujii T
  • Journal Title: Oncotarget Volume: 7 Issue: 20 Pages: 29577-29591
  • DOI: 10.18632/oncotarget.8807
  • NAID: 120005820854
  • Peer Reviewed / Open Access
• [Journal Article] Visualization of Neuregulin 1 ectodomain shedding reveals its local processing in vitro and in vivo. (2016)
  • Author(s): Kamezaki A, Sato F, Aoki K, Asakawa K, Kawakami K, Matsuzaki F, Sehara-Fujiwara A.
  • Journal Title: Scientific Reports Volume: 6 Issue: 1 Pages: 28873-28873
  • DOI: 10.1038/srep28873
  • NAID: 120005819488
  • Peer Reviewed / Open Access
• [Journal Article] Two New FRET Imaging Measures: Linearly Proportional to and Highly Contrasting the Fraction of Active Molecules. (2016)
  • Author(s): Yamao M, Aoki K, Yukinawa N, Ishii S, Matsuda M, Naoki H.
  • Journal Title: PLoS One Volume: 11 Issue: 10 Pages: e0164254-e0164254
  • DOI: 10.1371/journal.pone.0164254
  • NAID: 120005853281
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Variegated RHOA mutations in adult T-cell leukemia/lymphoma (2016)
  • Author(s): Nagata Y, Kontani K, Enami T, Kataoka K, Ishii R, Totoki Y, Kataoka TR, Hirata M, Aoki K, Nakano K, Kitanaka A, Sakata-Yanagimoto M, Egami S, Shiraishi Y, Chiba K, Tanaka H, Shiozawa Y, Yoshizato T, (9名), Miyano S, Nureki O, Shibata T, Haga H, Shimoda K, Katada T, Chiba S, Watanabe T, Ogawa S
  • Journal Title: Blood Volume: 127 Issue: 5 Pages: 596-604
  • DOI: 10.1182/blood-2015-06-644948
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Synergistic antitumor effects of combination of PI3K/mTOR and MEK inhibition (SAR245409 and pimasertib) in mucinous ovarian carcinoma cells by fluorescence resonance energy transfer imaging. (2016)
  • Author(s): naba K, Oda K, Aoki K, Sone K, Ikeda Y, Miyasaka A, Kashiyama T, Fukuda T, Makii C, Arimoto T, Wada-Hiraike O, Kawana K, Yano T, Osuga Y, Fujii T.
  • Journal Title: Oncotarget Volume: 未定
  • NAID: 120005820854
  • Peer Reviewed
• [Journal Article] mTORC1 upregulation via ERK-dependent gene expression change confers intrinsic resistance to MEK inhibitors in oncogenic KRas-mutant cancer cells. (2015)
  • Author(s): Komatsu N, Fujita Y, Matsuda M, Aoki K
  • Journal Title: Oncogene Volume: in press Issue: 45 Pages: 5607-5616
  • DOI: 10.1038/onc.2015.16
  • NAID: 120005712924
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Intercellular propagation of extracellular signal-regulated kinase activation revealed by in vivo imaging of mouse skin. (2015)
  • Author(s): Hiratsuka T, Fujita Y, Naoki H, Aoki K, Kamioka Y, Matsuda M
  • Journal Title: eLife Volume: - Pages: 1-18
  • DOI: 10.7554/elife.05178
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Quantitative in vivo fluorescence cross-correlation analyses highlight the importance of competitive effects in the regulation of protein-protein interactions (2014)
  • Author(s): Sadaie W, Harada Y, Matsuda M, Aoki K
  • Journal Title: Molecular and Cellular Biology Volume: 34 Issue: 17 Pages: 3272-3290
  • DOI: 10.1128/mcb.00087-14
  • NAID: 120005625632
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Fluorescence resonance energy transfer based quantitative analysis of feedforward and feedback loops in epidermal growth factor receptor signaling and the sensitivity to molecular targeting drugs. (2014)
  • Author(s): Fujita, Y., Komatsu, N., Matsuda, M., and Aoki, K
  • Journal Title: The FEBS journal Volume: 281 Issue: 14 Pages: 3177-3192
  • DOI: 10.1111/febs.12852
  • NAID: 120005647503
  • Peer Reviewed / Open Access
• [Presentation] 実験と数理・統計を組み合わせた癌研究 (2016)
  • Author(s): 青木 一洋
  • Organizer: 第1回肺癌バイオカンファレンス
  • Place of Presentation: 日本ベーリンガーインゲルハイム株式会社17FBoadRoom（東京都品川区）
  • Invited
• [Presentation] Synthetic lethal screening with the MEK inhibitor Trametinib in KRAS-mutant cancer cells (2015)
  • Author(s): Aoki K
  • Organizer: 第74回日本がん学会学術総会
  • Place of Presentation: 名古屋国際会議場
  • Year and Date: 2015-10-10
• [Presentation] FRETイメージングによるEGFシグナル伝達系のフィードフォワード・フィードバック経路の定量解析と分子標的薬感受性の予測 (2014)
  • Author(s): 青木 一洋
  • Organizer: 第87回日本生化学会年会
  • Place of Presentation: 国立京都国際会館
  • Year and Date: 2014-10-15
• [Presentation] イメージングによる細胞内シグナル伝達系と分子標的薬抵抗性の定量解析 (2014)
  • Author(s): 青木 一洋
  • Organizer: 第11回日本病理学会カンファレンス
  • Place of Presentation: 六甲山ホテル
  • Year and Date: 2014-08-01 – 2014-08-02
  • Invited
• [Presentation] FCCSによるin vivo Kdの定量法の開発 (2014)
  • Author(s): 青木 一洋
  • Organizer: 第66回日本細胞生物学会
  • Place of Presentation: 奈良県新公会堂
  • Year and Date: 2014-06-13
  • Invited
• [Presentation] Imaging and Mathematical Modeling of Molecular Activities in Living Cells (2014)
  • Author(s): Kazuhiro Aoki
  • Organizer: 2014 Japan-America Frontiers of Engineering Symposium
  • Place of Presentation: National Museum of Emerging Science and Innovation (Miraikan)
  • Year and Date: 2014-06-09 – 2014-06-11
  • Invited
• [Presentation] 画像解析による多細胞動態の解析 (2014)
  • Author(s): 青木 一洋
  • Organizer: 第47回日本発生生物学会年会
  • Place of Presentation: ウィンク愛知
  • Year and Date: 2014-05-27
  • Invited
• [Presentation] イメージングによる細胞内シグナル伝達系の定量解析 (2014)
  • Author(s): 青木 一洋
  • Organizer: Research PlaNet 2014
  • Place of Presentation: 東京国際フォーラム
  • Year and Date: 2014-05-10 – 2014-05-11
  • Invited
• [Book] バイオ画像解析 手とり足とり (2014)
  • Author(s): 小林徹也、青木一洋
  • Total Pages: 220
  • Publisher: 羊土社
• [Remarks] 蛍光相互相関分光法による細胞内解離定数の測定法の開発
  • URL: https://sites.google.com/site/qsimulationproject/project-definition/invivokd
• [Remarks] フィードフォワードとフィードバック制御 および分子標的薬の感受性についての定量解析
  • URL: https://sites.google.com/site/qsimulationproject/project-definition/fb