Cell Reports (Oct 2017)
A RhoA-FRET Biosensor Mouse for Intravital Imaging in Normal Tissue Homeostasis and Disease Contexts
- Max Nobis,
- David Herrmann,
- Sean C. Warren,
- Shereen Kadir,
- Wilfred Leung,
- Monica Killen,
- Astrid Magenau,
- David Stevenson,
- Morghan C. Lucas,
- Nadine Reischmann,
- Claire Vennin,
- James R.W. Conway,
- Alice Boulghourjian,
- Anaiis Zaratzian,
- Andrew M. Law,
- David Gallego-Ortega,
- Christopher J. Ormandy,
- Stacey N. Walters,
- Shane T. Grey,
- Jacqueline Bailey,
- Tatyana Chtanova,
- Julian M.W. Quinn,
- Paul A. Baldock,
- Peter I. Croucher,
- Juliane P. Schwarz,
- Agata Mrowinska,
- Lei Zhang,
- Herbert Herzog,
- Andrius Masedunskas,
- Edna C. Hardeman,
- Peter W. Gunning,
- Gonzalo del Monte-Nieto,
- Richard P. Harvey,
- Michael S. Samuel,
- Marina Pajic,
- Ewan J. McGhee,
- Anna-Karin E. Johnsson,
- Owen J. Sansom,
- Heidi C.E. Welch,
- Jennifer P. Morton,
- Douglas Strathdee,
- Kurt I. Anderson,
- Paul Timpson
Affiliations
- Max Nobis
- The Garvan Institute of Medical Research, St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia
- David Herrmann
- The Garvan Institute of Medical Research, St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia
- Sean C. Warren
- The Garvan Institute of Medical Research, St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia
- Shereen Kadir
- Cancer Research UK Beatson Institute, Switchback Road, Bearsden, Glasgow G611BD, UK
- Wilfred Leung
- The Garvan Institute of Medical Research, St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia
- Monica Killen
- The Garvan Institute of Medical Research, St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia
- Astrid Magenau
- The Garvan Institute of Medical Research, St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia
- David Stevenson
- Cancer Research UK Beatson Institute, Switchback Road, Bearsden, Glasgow G611BD, UK
- Morghan C. Lucas
- The Garvan Institute of Medical Research, St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia
- Nadine Reischmann
- The Garvan Institute of Medical Research, St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia
- Claire Vennin
- The Garvan Institute of Medical Research, St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia
- James R.W. Conway
- The Garvan Institute of Medical Research, St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia
- Alice Boulghourjian
- The Garvan Institute of Medical Research, St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia
- Anaiis Zaratzian
- The Garvan Institute of Medical Research, St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia
- Andrew M. Law
- The Garvan Institute of Medical Research, St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia
- David Gallego-Ortega
- The Garvan Institute of Medical Research, St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia
- Christopher J. Ormandy
- The Garvan Institute of Medical Research, St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia
- Stacey N. Walters
- The Garvan Institute of Medical Research, St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia
- Shane T. Grey
- The Garvan Institute of Medical Research, St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia
- Jacqueline Bailey
- The Garvan Institute of Medical Research, St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia
- Tatyana Chtanova
- The Garvan Institute of Medical Research, St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia
- Julian M.W. Quinn
- The Garvan Institute of Medical Research, St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia
- Paul A. Baldock
- The Garvan Institute of Medical Research, St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia
- Peter I. Croucher
- The Garvan Institute of Medical Research, St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia
- Juliane P. Schwarz
- Cancer Research UK Beatson Institute, Switchback Road, Bearsden, Glasgow G611BD, UK
- Agata Mrowinska
- Cancer Research UK Beatson Institute, Switchback Road, Bearsden, Glasgow G611BD, UK
- Lei Zhang
- The Garvan Institute of Medical Research, St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia
- Herbert Herzog
- The Garvan Institute of Medical Research, St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia
- Andrius Masedunskas
- Neuromuscular and Regenerative Medicine Unit, University of New South Wales, Sydney, NSW 2010, Australia
- Edna C. Hardeman
- Neuromuscular and Regenerative Medicine Unit, University of New South Wales, Sydney, NSW 2010, Australia
- Peter W. Gunning
- Oncology Research Unit, School of Medical Sciences, University of New South Wales, Sydney, NSW 2010, Australia
- Gonzalo del Monte-Nieto
- Developmental and Stem Cell Biology Division, Victor Chang Cardiac Research Institute, Sydney, NSW 2010, Australia
- Richard P. Harvey
- Developmental and Stem Cell Biology Division, Victor Chang Cardiac Research Institute, Sydney, NSW 2010, Australia
- Michael S. Samuel
- Centre for Cancer Biology, SA Pathology and University of South Australia School of Medicine, University of Adelaide, Adelaide, SA 5000, Australia
- Marina Pajic
- The Garvan Institute of Medical Research, St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia
- Ewan J. McGhee
- Cancer Research UK Beatson Institute, Switchback Road, Bearsden, Glasgow G611BD, UK
- Anna-Karin E. Johnsson
- Signalling Programme, Babraham Institute, Cambridge CB223AT, UK
- Owen J. Sansom
- Cancer Research UK Beatson Institute, Switchback Road, Bearsden, Glasgow G611BD, UK
- Heidi C.E. Welch
- Signalling Programme, Babraham Institute, Cambridge CB223AT, UK
- Jennifer P. Morton
- Cancer Research UK Beatson Institute, Switchback Road, Bearsden, Glasgow G611BD, UK
- Douglas Strathdee
- Cancer Research UK Beatson Institute, Switchback Road, Bearsden, Glasgow G611BD, UK
- Kurt I. Anderson
- Francis Crick Institute, London NW11AT, UK
- Paul Timpson
- The Garvan Institute of Medical Research, St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia
- DOI
- https://doi.org/10.1016/j.celrep.2017.09.022
- Journal volume & issue
-
Vol. 21,
no. 1
pp. 274 – 288
Abstract
The small GTPase RhoA is involved in a variety of fundamental processes in normal tissue. Spatiotemporal control of RhoA is thought to govern mechanosensing, growth, and motility of cells, while its deregulation is associated with disease development. Here, we describe the generation of a RhoA-fluorescence resonance energy transfer (FRET) biosensor mouse and its utility for monitoring real-time activity of RhoA in a variety of native tissues in vivo. We assess changes in RhoA activity during mechanosensing of osteocytes within the bone and during neutrophil migration. We also demonstrate spatiotemporal order of RhoA activity within crypt cells of the small intestine and during different stages of mammary gestation. Subsequently, we reveal co-option of RhoA activity in both invasive breast and pancreatic cancers, and we assess drug targeting in these disease settings, illustrating the potential for utilizing this mouse to study RhoA activity in vivo in real time.
Keywords
- intravital imaging
- pancreatic cancer
- breast cancer
- actin
- small GTPase RhoA
- FLIM-FRET
- biosensors
- immunology
- development
- cell biology
