FLIM-FRET imaging in vivo reveals 3D-environment spatially regulates RhoGTPase activity during cancer cell invasion

McGhee, E. J., Morton, J. P. , Von Kriegsheim, A., Schwarz, J. P., Karim, S. A., Carragher, N. O., Sansom, O. , Anderson, K. I. and Timpson, P. (2011) FLIM-FRET imaging in vivo reveals 3D-environment spatially regulates RhoGTPase activity during cancer cell invasion. Small GTPases, 2(4), pp. 239-244. (doi: 10.4161/sgtp.2.4.17275) (PMID:22145098) (PMCID:PMC3225915)

Abstract

Many conceptual advances in biology have been achieved by experimental studies using planar two-dimensional cell culture systems. Recent adaptations of molecular techniques to three-dimensional model systems are bridging the gap in our understanding of biological events in vitro and in vivo in the study of disease progression. Recently, in vitro studies using Förster resonance energy transfer (FRET) have shown that the prototypical RhoGTPases Cdc42, Rac and RhoA are temporally and spatially synchronized during cell migration, with initial RhoA activity inducing protrusion prior to activation of Rac. This simultaneous FRET approach illustrates the tight control and dynamic regulation of RhoGTPase activity necessary for coordinated cell migration in vitro. Here, we discuss our recent work using FLIM-FRET analysis in a three-dimensional setting to reveal another layer of regulation in which RhoA activity is governed by the extracellular microenvironment. We demonstrate that RhoA is spatially regulated into discrete fractions of activity at the leading edge and rear of cells during invasion in vivo or within three-dimensional matrices. Significantly, this spatial regulation of RhoA was absent in two-dimensional in vitro settings. This distinct sub-cellular regulation of RhoA at the poles of invading cells in three-dimensions sets a precedent that other RhoGTPases or signaling proteins may also be differentially regulated in a context-dependent manner during key biological processes such as invasion.

• Item Type: Articles
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Morton, Professor Jen and Anderson, Professor Kurt and McGhee, Dr Ewan and Timpson, Dr Paul and Von Kriegsheim, Mr Alexander and Karim, Ms Saadia and Sansom, Professor Owen
• Authors: McGhee, E. J., Morton, J. P., Von Kriegsheim, A., Schwarz, J. P., Karim, S. A., Carragher, N. O., Sansom, O., Anderson, K. I., and Timpson, P.
• College/School: College of Medical Veterinary and Life Sciences > School of Cancer Sciences
• Journal Name: Small GTPases
• Publisher: Landes Bioscience
• ISSN: 2154-1248
• ISSN (Online): 2154-1256
• Published Online: 01 August 2011