Supplementary Materials

Supplementary Material for:

Transient tissue priming via ROCK inhibition uncouples pancreatic cancer progression, sensitivity to chemotherapy, and metastasis

Claire Vennin, Venessa T. Chin, Sean C. Warren, Morghan C. Lucas, David Herrmann, Astrid Magenau, Pauline Melenec, Stacey N. Walters, Gonzalo del Monte-Nieto, James R. W. Conway, Max Nobis, Amr H. Allam, Rachael A. McCloy, Nicola Currey, Mark Pinese, Alice Boulghourjian, Anaiis Zaratzian, Arne A. S. Adam, Celine Heu, Adnan M. Nagrial, Angela Chou, Angela Steinmann, Alison Drury, Danielle Froio, Marc Giry-Laterriere, Nathanial L. E. Harris, Tri Phan, Rohit Jain, Wolfgang Weninger, Ewan J. McGhee, Renee Whan, Amber L. Johns, Jaswinder S. Samra, Lorraine Chantrill, Anthony J. Gill, Maija Kohonen-Corish, Richard P. Harvey, Andrew V. Biankin; Australian Pancreatic Cancer Genome Initiative (APGI), T. R. Jeffry Evans, Kurt I. Anderson, Shane T. Grey, Christopher J. Ormandy, David Gallego-Ortega, Yingxiao Wang, Michael S. Samuel, Owen J. Sansom, Andrew Burgess, Thomas R. Cox, Jennifer P. Morton, Marina Pajic*, Paul Timpson*

*Corresponding author. Email: m.pajic{at}garvan.org.au (M. Pajic); p.timpson{at}garvan.org.au (P.T.)

Published 5 April 2017, Sci. Transl. Med. 9, eaai8504 (2017)
DOI: 10.1126/scitranslmed.aai8504

This PDF file includes:

  • Materials and Methods
  • Fig. S1. ROCK inhibition with Fasudil and Y-27632 impairs ECM integrity.
  • Fig. S2. The CDK1-FRET biosensor distinguishes changes in CDK1 activity and is a surrogate for cell cycle arrest.
  • Fig. S3. Priming with Fasudil disrupts ECM remodeling, inhibits ROCK signaling, and improves Gem/Abraxane efficacy in vivo.
  • Fig. S4. Priming with Fasudil influences liver vasculature, cell attachment to CDMs, response to chemotherapy, and remodeling of the ECM.
  • Fig. S5. Priming with Fasudil results in decreased SRC activity and defects in mitosis.
  • Fig. S6. High ECM TKCC5 patient model responds to priming strategies in a 3D patient-personalized organotypic matrix.
  • Fig. S7. Low ECM TKCC2 patient model does not respond to priming strategies in a 3D patient-personalized organotypic matrix.
  • Fig. S8. TKCC5 orthotopic tumors respond to priming, and SHG does not predict survival.
  • Fig. S9. Priming with Fasudil uncouples PC progression.
  • Table S1. List of P values.
  • Table S2. Details of antibodies used for the study.
  • Legends for movies S1 to S9
  • Reference (107)

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Other Supplementary Material for this manuscript includes the following:

  • Movie S1 (.mp4 format). 4D monitoring of fibroblast-ECM interactions upon treatment with vehicle and Fasudil.
  • Movie S2 (.mp4 format). Live FLIM-FRET imaging of the CDK1 biosensor in KPC cells in response to Abraxane and Abraxane + RO3306.
  • Movie S3 (.mp4 format). Live FLIM-FRET imaging of the CDK1 biosensor in KPC cells in interphase and mitosis.
  • Movie S4 (.mp4 format). Live FLIM-FRET monitoring of CDK1 activity in KPC cells actively interacting with an organotypic matrix.
  • Movie S5 (.avi format). Intravital FLIM-FRET imaging of subcutaneous xenografts with KPC-CDK1 cells and imaging of fibrillar collagen.
  • Movie S6 (.mp4 format). Intravital monitoring of CDK1 accumulation in subcutaneous KPC tumors.
  • Movie S7 (.mp4 format). Intravital imaging of quantum dots circulating through tumor-associated vasculature and diffusing into tumor tissue upon priming with Fasudil.
  • Movie S8 (.mp4 format). Imaging of liver tissue with metastatic KPC cells expressing the CDK1 biosensor forming macrometastases and micrometastases.
  • Movie S9 (.mp4 format). Time-lapse tracking of collective cell streaming on CDMs unprimed or primed with Fasudil.