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International audience; Glioblastomas are the most frequent adult primary brain tumors that still remain fatal despite major clinical efforts. As in other solid tumors, populations of glioblastoma stem-like cells (GSCs) endowed with tumor initiating and therapeutic resistance properties have been identified. Glioblastomas are highly vascularized tumors resulting in a rich dialog between GSCs and endothelial cells. In one direction, endothelial cells and their secreted proteins are able to sustain GSC properties while, in turn, GSCs can promote neoangiogenesis, modulate endothelial cell functions and may even transdifferentiate into endothelial cells. Accordingly, targeting tumor vasculature seems a promising issue despite incomplete and transient results obtained from anti-vascular endothelial growth factor therapeutic trials. Recent findings of novel GSC-secreted molecules with pro-angiogenic properties (Semaphorin 3A, hepatoma-derived growth factor) open the path to the design of a concerted attack of glioblastoma vasculature that could overcome the development of resistance to single-targeted therapies while keeping away the toxicity of the treatments.

Numerous cell surface receptors, including tyrosine kinase and G protein-coupled receptors, play critical roles in endothelial cell function and blood vessel development. These receptors share the ability of stimulating an intricate network of intracellular signaling pathways, including the activation of members of the Ras and Rho family of small GTPases. However, the contribution of these signaling molecules to the numerous biological activities performed by endothelial cells is still not fully understood. Here, we have used a conditional Cre/Flox approach, enabling the deletion of the Rac1 gene in endothelial cells, to examine the role of the Rho-related GTPase Rac1 in endothelial cell function and vascular development. Rac1 excision in primary endothelial cells in vitro revealed that Rac1 plays a central role in endothelial cell migration, tubulogenesis, adhesion, and permeability in response to vascular endothelial growth factor (VEGF) and sphingosine-1-phosphate (S1P), which is likely due to the inability of Rac1-deficient endothelial cells to form lamellipodial structures and focal adhesions, and to remodel their cell-cell contacts. Importantly, endothelial-specific excision of Rac1 results in embryonic lethality in midgestation (around E9.5), and defective development of major vessels and complete lack of small branched vessels was readily observed in these endothelial Rac1-deficient embryos and their yolk sacs. These findings provide direct evidence that the activity of Rac1 in endothelial cells is essential for vascular development and suggest that Rac1 and its downstream targets may represent promising therapeutic targets for the treatment of numerous human diseases that involve aberrant neovascularization.

Melanoma cell adhesion molecule (MCAM), an adhesion molecule belonging to the Ig superfamily, is an endothelial marker and is expressed in different epithelia. MCAM is expressed as two isoforms differing by their cytoplasmic domain: MCAM-l and MCAM-s (long and short). In order to identify the respective role of each MCAM isoform, we analyzed MCAM isoform targeting in polarized epithelial Madin–Darby canine kidney (MDCK) cells using MCAM-GFP chimeras. Confocal microscopy revealed that MCAM-s and MCAM-l were addressed to the apical and basolateral membranes, respectively. Transfection of MCAM-l mutants established that a single dileucine motif (41-42) of the cytoplasmic domain was required for MCAM-l basolateral targeting in MDCK cells. Although double labelling experiments showed that MCAM-l is not a component of adherens junctions and focal adhesions, its expression on basolateral membranes suggests that MCAM-l is involved in epithelium insuring.

beta-Catenin was initially characterized as a protein interacting with the cadherin cytoplasmic tail and regulating cell-cell contacts and actin cytoskeleton interactions. Moreover, the gene coding for the Drosophila orthologue of beta-catenin, armadillo, was independently identified downstream of wingless in the segment-polarity signalling pathway. In fact, beta-catenin/Armadillo turned out to be key mediators of the Wnt/Wingless pathways in vertebrates and invertebrates. beta-Catenin participates in both adhesion and signalling functions in a mutually exclusive manner; bound to cadherins at the plasma membrane or 'unbound' in cytosolic or nuclear complexes. This model had placed beta-catenin at the crossroads between cadherin and Wnt signalling, leading to the dogma of inhibition of beta-catenin signalling by cadherins.

The physiological role of the prion protein is largely unknown. Here, clustering of prion at the surface of GT1-7 cells was observed upon anti-prion antibody treatments. This clustering was associated with a rapid and transient phosphorylation of the mitogen activated protein kinases (MAPKs) extracellular receptor kinases 1 and 2 (ERK1/2), and also of the microtubule-destabilizing protein stathmin at serine 16. The specificity of this antibody-mediated activation was ascertained by its inhibition by prion small interfering RNA. The phosphorylation of ERK1/2 but not that of stathmin was abolished by the MAPK/ERK kinase 1 inhibitor U0126, whereas both signaling pathways were blocked by the specific inhibitor of the epidermal growth factor receptor AG1478, suggesting the likely recruitment of this receptor upon prion clustering.