IMAGING G PROTEINS IN CELL ADHESION AND MIGRATION
Our research focuses in understanding the roles of the G protein-couple receptors (GPCRs)and their mechanisms of action in inducing the migration of cells and disruption of cell adhesion. Both processes are important to lead cells to metastasis and cancer.GPCRs are the largest family of molecules located in the surface of the cell and responsible for transmitting signal from outside to the inside of the cell. Recently some of these receptors have emerged as crucial players in tumor growth and metastasis. Malignant cells often hijack the normal functions of these receptors in order to survive, proliferate autonomously and invade their surrounding tissues. Our group investigates the mechanism of action of one GPCR receptor, the chemokine receptor CCR2. This receptor is involved in different diseases associated with inflammation as well as in processes like cancer. Our goal is to discern to its mechanism of action. For this during these two years we have been studying the molecules associated with its activation. We have accomplished the main goals of the second year of the project: i/ we have finished the analysis of the chemokine receptor and filamin; ii/ we have performed a through study by confocal microscopy of the localization of filamin and CCR2B; iii/ we have found that filamin A protein is required for efficient internalization of the receptor in the cell; iv/ a manuscript was sent for publication to PNAS on June 2009, though they have good comments (answer came back on September 2009) they suggested us to send it to a more specialized journal. v/ we have now performed and finished some extra control experiments. We have rewritten the manuscript and it will be sent for publication during February 2010. vi/ At the same time, we have started the study of chemokine signaling and filamin interaction in two monocytic human cell lines: Mono Mac cells and THP-1; vii/ we have evidence that filamin A is needed for differentiation of monocytes to macrophages. We expect that during this last year we will finish another article on the subject of CCR2B and FLNA and that Mr. Minsaas will be presenting her Thesis during 2010. We also plan to sent another paper for publication within the same year.
Our research focuses in understanding how G protein signaling can affect the transition between cell adhesion and migration that can lead to metastasisG protein-couple receptors (GPCRs), the largest family of cell surface molecules involved in signal transmission, have recently emerged as crucial players in tumor growth and metastasis. Malignant cells often hijack the normal physiological functions of GPCRs to survive, proliferate autonomously, evade immune system, increase their blood supply, invade their surrounding tissues and disseminate their organs. Our research focuses in understanding the roles of G proteins and their signaling in cell migration and disruption of cell adhesion, both processes are important to lead cells to metastasis. The three years project approved from Helse Vest last year has two working fronts: i/ the study of chemokines in cell migration ii/ the study of G proteins (G12) and disruption of cell adhesion We have accomplished the main goals of the first year of the project: chemokines, i/ we have finished the in vitro analysis of the chemokine receptor and filamin; ii/ we have started our studies with confocal microscopy of the localization of filamin and CCR2B in knock-out cells; iii/ we have finished the first run of experiments for the filamin-CCR2B project and a manuscript is being written; iv/ we have constructed a kimeric filamin protein with GFP in the hinge region 1; v/ we have started the experiments to discern the mecanism of internalization and the involvement of filamin. G12 proteins and cadherins, i/ we have analyzed the phosphorylation of p120ctn in presence of G12; ii/ we have performed FRAP experiments with p120ctn; iii/ we have advanced in our studies in the imaging of cell migration. Dr. Panaguma, expert in image techniques has been an important collaborator for the outcome of the results on filamin. Ms Vandana is the key contributor to our advances in the study of G12 proteins. An manuscript has been written and it will be sent for publication January 2009, with the results of the project of filamin: “Binding of the CCR2B receptor to Filamin A regulates receptor internalization and recycling”. Authors: Laura Minsaas, Jesus Planagumà, Arieh Katz and Anna M. Aragay.
Laura Minsaas, Jesus Planagumà, Arieh Katz and Anna M. Aragay
Binding of the CCR2B receptor to Filamin A regulates receptor internalization and recycling
It is sent on January 2009