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 BaileyTatyana 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

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49 Citations (Scopus)

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. Nobis et al. generated a RhoA-FRET biosensor mouse to characterize and quantify the spatiotemporal distribution of RhoA activity in native mammalian tissues in vivo during development and disease progression. They show that RhoA activity is tightly regulated during various normal biological processes and is co-opted in disease settings, such as invasive breast and pancreatic cancers.

Original languageEnglish
Pages (from-to)274-288
Number of pages15
JournalCell Reports
Volume21
Issue number1
DOIs
Publication statusPublished - 3 Oct 2017
Externally publishedYes

Keywords

  • actin
  • biosensors
  • breast cancer
  • cell biology
  • development
  • FLIM-FRET
  • immunology
  • intravital imaging
  • pancreatic cancer
  • small GTPase RhoA

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