cytoskeleton; FRET biosensors; Signaling; Microfluidics; cell migration
Martin Katrin, Reimann Andreas, Fritz Rafael D, Ryu Hyunryul, Jeon Noo Li, Pertz Olivier (2016), Spatio-temporal co-ordination of RhoA, Rac1 and Cdc42 activation during prototypical edge protrusion and retraction dynamics., in Scientific reports
, 6, 21901-21901.
Fritz Rafael Dominik, Pertz Olivier (2016), The dynamics of spatio-temporal Rho GTPase signaling: formation of signaling patterns., in F1000Research
, 5, 1-3.
Moretti Francesca, Rolando Chiara, Winker Moritz, Ivanek Robert, Rodriguez Javier, Von Kriegsheim Alex, Taylor Verdon, Bustin Michael, Pertz Olivier (2015), Growth Cone Localization of the mRNA Encoding the Chromatin Regulator HMGN5 Modulates Neurite Outgrowth., in Molecular and cellular biology
, 35(11), 2035-50.
Fritz Rafael Dominik, Menshykau Denis, Martin Katrin, Reimann Andreas, Pontelli Valeria, Pertz Olivier (2015), SrGAP2-Dependent Integration of Membrane Geometry and Slit-Robo-Repulsive Cues Regulates Fibroblast Contact Inhibition of Locomotion., in Developmental cell
, 35(1), 78-92.
Martin Katrin, Vilela Marco, Jeon Noo Li, Danuser Gaudenz, Pertz Olivier (2014), A growth factor-induced, spatially organizing cytoskeletal module enables rapid and persistent fibroblast migration., in Developmental cell
, 30(6), 701-16.
Fritz Rafael, Letzelter Michel, Reimann Andreas, Martin Katrin, Fusco Ludovico, Ritsma Laila, Ponsionen B, Fluri Erika, Schulte-Merker Stefan, Van Rheenen Jacco, Pertz Olivier (2014), A versatile toolkit to produce sensitive FRET biosensors to visualize signaling in time and space., in Science Signaling
, 2(285), 1-7.
Feltrin D., Pertz O. (2012), Assessment of Rho GTPase signaling during neurite outgrowth, in Francesco Rivero (ed.), Humana Press, England, 181-94.
Feltrin D., Fusco L., Witte H., Moretti F., Martin K., Letzelter M., Fluri E., Scheiffele P., Pertz O. (2012), Growth Cone MKK7 mRNA Targeting Regulates MAP1b-Dependent Microtubule Bundling to Control Neurite Elongation, in PLoS Biol
, 10, 1001439-1001439.
The purpose of this grant proposal is to study spatio-temporal Rho GTPase signaling mechanisms to the cytoskeleton during cell migration. An essential emphasis will be to study these signaling events at the spatial and temporal scales at which they are occurring (e.g. single microns and tens of seconds respectively). This will take advantage of the novel tools that were developed in the framework of the previous SNF grant proposal. Using 1. a novel generation of FRET-based sensors that monitor Rho GTPase activation, and 2. microfluidic platforms that allow to induce standardized, robust cell migration behaviors, we propose to study spatio-temporal Rho GTPase signaling programs during two robust prototypical cell migration systems:- haptokinesis (cell migration in response to integrin signaling)- PDGF chemokinesis (cell migration in response to stimulation with the soluble chemokine platelet derived growth factor).Our preliminary data shows that both cell migration systems use different actin and adhesion dynamics, that parallel different cell morphodynamics and lead to different migration speeds. The hypothesis relevant to this proposal is that this correlates with different Rho GTPase signaling programs. An important feature of the rapid PDGF chemokinesis cell migration mode is that it allows a high co-ordination between the leading edge and the retracting tail of the cell allowing for robust assessment how both processes are linked.Specifically, we will:- Study the dynamic behavior of a panel of cytoskeletal and adhesion markers in both cell migration modes.- Study the spatio-temporal patterns of RhoA, Rac1 and Cdc42 activity in both cell migration modes.This will then allow to merge these datasets to produce an integrated view of how spatio-temporal Rho GTPase signaling integrates with the different cytoskeletal and adhesion dynamics characteristic of each cell migration mode. This will also allow to put specific spatio-temporal Rho GTPase signaling events in a correct contextual background.We will then perturbate a set of Rac1 activators (GEFs) that are known to signal downstream of integrins or receptor tyrosine kinases (PDGF chemokinesis) and explore how this affects both cell migration modes. The hypothesis relevant to this aim is that these different GEFs regulate spatio-temporal pools of Rac1 with specific functions in time and space. This should again put each GEF in a specific context and extend previous models in which these GEFs has been studied independently in a large variety of cellular contexts.We foresee that this study will allow to significantly extend previous models of cell migration by providing an integrated view of how Rho GTPases regulate and are regulated by cytoskeletal and adhesion dynamics in two prototypical cell migration systems. This should provide a novel paradigm for spatio-temporal Rho GTPase signaling. A clear understanding of these signaling pathways might by example allow to specifically target a given cell migration mode and could be the starting point to design drugs for clinically relevant cell migration events. An example could be the cell migration events that occur during inflammation that results from PDGF-induced fibroblast recruitment to a wound during wound healing.