Functional intravital imaging of the breast tumor invasion process in tumor microenvironment

• Project/Area Number: 23701053
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Tumor biology
• Research Institution: Kyoto University
• Principal Investigator: KAMIOKA Yuji 京都大学, 医学(系)研究科(研究院), 助教 (50511424)
• Project Period (FY): 2011 – 2013
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2013: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2012: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000); Fiscal Year 2011: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
• Keywords: FRET / 蛍光共鳴エネルギー移動 / イメージング / 癌微小環境 / 生体イメージング / ERK / PKA / 低分子量Gタンパク質 / Rhoファミリー / 乳癌 / 国際情報交流

Research Abstract

A variety of fluorescent biosensors based on the principle of Fluorescence Resonance Energy Transfer (FRET) have been well used to visualize the activities of various signaling molecules in vitro. However, living tissues actually consist of many kinds of cells and cellular interactions in three-dimensional environments.
Recent studies show the importance of tumor microenvironment comprising of tumor cells and stroma cells. In this project, molecular activities of ERK, PKA and Rac1 were visualized by using genetically-encoded FRET biosensors in living mice under two-photon microscope for the understanding of tumor microenvironment in vivo. We have developed transgenic mice which express FRET biosensors for ERK and PKA in this project. In addition, imaging tools, stabilizer and windows were developed to get motion-less images and keep mice alive. These results achieved by intravital imaging can progress the studies about tumor microenvironment.

Research Products

• [Journal Article] Heterogeneity in ERK activity as visualized by in vivo FRET imaging or mammary tumor cells developed in MMTV-Neu mice. (2014)
  • Author(s): Yuka Kumagai, Honda Naoki, Eiji Nakasyo, Yuji Kamioka, Etsuko Kiyokawa, Michiyuki Matsuda
  • Journal Title: Oncogene Volume: (ネット上で先行掲載) Issue: 8 Pages: 1051-7
  • DOI: 10.1038/onc.2014.28
  • Peer Reviewed / Open Access
• [Journal Article] Cell cycle-dependent Rho GTPase activity dynamically regulates cancer cell motility and invasion in vivo. (2014)
  • Author(s): Kagawa Y, Matsumoto S, Kamioka Y, Mimori M, Naito Y, Ishii T, Okuzaki D, Nishida N, Maeda S, Naito A, Kikuta J, Nishikawa K, Nishimura J, Haraguchi N, Takemasa I, Mizushima T, Ikeda M, Yamamoto H, Sekimoto M, Ishii H, Doki Y, Matsuda M, Kikuchi A, Mori M, Ishii M
  • Journal Title: PLoS One Volume: 8 Issue: 12 Pages: e83629-e83629
  • DOI: 10.1371/journal.pone.0083629
  • Peer Reviewed / Open Access
• [Journal Article] Fluorescence resonance energy transfer imaging of cell signaling from in vitro to in vivo: Basis of biosensor construction, live imaging, and image processing (2013)
  • Author(s): Aoki, K.
  • Journal Title: Development Growth and Differentiation Volume: in press Issue: 4 Pages: 515-522
  • DOI: 10.1111/dgd.12039
  • NAID: 40019735187
  • Peer Reviewed
• [Journal Article] GDNF and endothelin 3 regulate migration of enteric neural crest-derived cells via PKA and Rac1 (2013)
  • Author(s): Goto A, Sumiyama K, Kamioka Y, Nakasho E, Ito K, Iwasaki M, Enomoto H, Matsuda M.
  • Journal Title: Journal of Neuroscience Volume: 33 Issue: 11 Pages: 4901-4912
  • DOI: 10.1523/jneurosci.4828-12.2013
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Live Imaging of Protein Kinase Activities in Transgenic Mice Expressing FRET Biosensors (2012)
  • Author(s): Kamioka, Y., Sumiyama, K., Mizuno, R., Sakia, Y., Hirata, E., Kiyokawa, E., Matsuda, M.
  • Journal Title: Cell Structure and Function Volume: 37 Issue: 1 Pages: 65-73
  • DOI: 10.1247/csf.11045
  • NAID: 130004137578
  • ISSN: 0386-7196, 1347-3700
  • Peer Reviewed
• [Journal Article] Live Imaging of Protein Kinase Activities in Transgenic Mice Expressing FRET Biosensors (2012)
  • Author(s): Yuji Kamioka, Kenta Sumiyama, Rei Mizuno, Yoshiharu Sakai, Eishu Hirata, Etsuko Kiyokawa and Michiyuki Matsuda
  • Journal Title: Cell Structure and Function Volume: 37 Pages: 66-73
  • NAID: 40019581951
  • Peer Reviewed
• [Presentation] Visualization of ERK and PKA activities in the tumor microenvironment consisting of lung-metastasized 4T1 murine breast tumor cells (2014)
  • Author(s): Yuji Kamioka and Michiyuki Matsuda
  • Organizer: An AACR Special Conference Cellular Heterogeneity in the Tumor Microenvironment （ポスター発表）
  • Place of Presentation: San Diego, US
• [Presentation] Live Imaging of Transgenic Mice Expressing FRET Biosensors (2013)
  • Author(s): Yuji Kamioka, Kenta Sumiyama, Rei Mizuno, Michiyuki Matsuda
  • Organizer: 35th Annual International Conference of IEEE Engineering in Medicine and Biology Society （口頭発表）
  • Place of Presentation: 大阪
  • Invited
• [Presentation] Live Imaging of Protein Kinase Activitiesin Transgenic Mice(口頭発表、ポスター発表) (2012)
  • Author(s): 上岡裕治
  • Organizer: 第45回日本発生生物学会・第64回日本細胞生物学会合同大会
  • Place of Presentation: 神戸
  • Year and Date: 2012-05-31
• [Presentation] Live Imaging of Protein Kinase Activities in Transgenic Mice (2012)
  • Author(s): Yuji Kamioka, Kenta Sumiyama , Rei Mizuno and Michiyuki Matsuda
  • Organizer: Joint Meeting of The 45th Annual Meeting of the Japanese Society of Developmental Biologists & The 64th Annual Meeting of the Japan Society for Cell Biology
  • Place of Presentation: 神戸
• [Presentation] Live Imaging of Protein Kinase Activities in Transgenic Mice (2012)
  • Author(s): Yuji Kamioka, Kenta Sumiyama , Rei Mizuno and Michiyuki Matsuda
  • Organizer: The 4th EMBO meeting NICE
  • Place of Presentation: Nice, France
• [Remarks]
  • URL: http://www.lif.kyoto-u.ac.jp/labs/fret/
• [Remarks] 京都大学大学院 医学研究科 病態生物医学講座
  • URL: http://www.lif.kyoto-u.ac.jp/labs/fret/
• [Remarks] 京都大学大学院 生命科学研究科・生体制御学 医学研究科・病態生物医学 松田研究室
  • URL: http://www.lif.kyoto-u.ac.jp/labs/fret/