Nucleus-invadopodia duo during cancer invasion 1

International audienceMatrix proteolysis mediated by MT1-MMP facilitates the invasive migration of tumor cells in dense tissues, which otherwise get trapped in the matrix because of limited nuclear deformability. A digest-on-demand response has been identified, which requires nucleus-microtubule linkage through the LINC complex and triggers MT1-1

La innovación tecnológica como catalizador de la recuperación post COVID-19 de las empresas agroalimentarias

La recuperación económica del sector agroalimentario ante la pandemia por COVID-19 presenta retos, entre ellos la reactivación del sector hotelero y de servicios de alimentación, la reconversión de nichos de mercado alimentarios premium y la consolidación de ventas en el mercado nacional; que deben ser asumidos con medidas que aseguren la inocuidad alimentaria, la integridad de los trabajadores y con la incorporación de innovaciones tecnológicas. Por lo tanto, los procesos de investigación e innovación son indispensables para responder a las necesidades del mercado, aumentar la competitividad y optimizar el uso de los recurso

Un substrat de micropiliers pour étudier la migration cellulaire

Les propriétés mécaniques des cellules jouent un rôle prépondérant dans de nombreux événements de la vie cellulaire comme le développement embryonnaire, la formation des tissus ou encore le développement des métastases. La migration cellulaire est en partie caractérisée par des interactions mécaniques. Ainsi, les forces de traction qu’exercent les cellules sur leur environnement impliquent, en parallèle, une réorganisation dynamique des processus d’adhérence et du cytosquelette interne de la cellule. Pour évaluer ces forces, un substrat a été développé, constitué d’un réseau forte densité de micro-piliers déformables sur lequel se déplacent les cellules. Cette surface est fabriquée par des méthodes de lithographie empruntées à la micro-électronique. Les piliers mesurent environ un micromètre et sont en caoutchouc, donc suffisamment déformables pour fléchir sous l’effet des forces exercées par les cellules. L’analyse au microscope des déflexions individuelles de chaque pilier a permis de quantifier en temps réel les forces locales que des cellules exercent sur leur substrat lors de leurs processus d’adhérence et de dissociation.Mechanical forces play an important role in various cellular functions, such as tumor metastasis, embryonic development or tissue formation. Cell migration involves dynamics of adhesive processes and cytoskeleton remodelling, leading to traction forces between the cells and their surrounding extracellular medium. To study these mechanical forces, a number of methods have been developed to calculate tractions at the interface between the cell and the substrate by tracking the displacements of beads or microfabricated markers embedded in continuous deformable gels. These studies have provided the first reliable estimation of the traction forces under individual migrating cells. We have developed a new force sensor made of a dense array of soft micron-size pillars microfabricated using microelectronics techniques. This approach uses elastomeric substrates that are micropatterned by using a combination of hard and soft lithography. Traction forces are determined in real time by analyzing the deflections of each micropillar with an optical microscope. Indeed, the deflection is directly proportional to the force in the linear regime of small deformations. Epithelial cells are cultured on our substrates coated with extracellular matrix protein. First, we have characterized temporal and spatial distributions of traction forces of a cellular assembly. Forces are found to depend on their relative position in the monolayer : the strongest deformations are always localized at the edge of the islands of cells in the active areas of cell protrusions. Consequently, these forces are quantified and correlated with the adhesion/scattering processes of the cells

ADP ribosylation factor 6 is activated and controls membrane delivery during phagocytosis in macrophages

Engulfment of particles by phagocytes is induced by their interaction with specific receptors on the cell surface, which leads to actin polymerization and the extension of membrane protrusions to form a closed phagosome. Membrane delivery from internal pools is considered to play an important role in pseudopod extension during phagocytosis. Here, we report that endogenous ADP ribosylation factor 6 (ARF6), a small GTP-binding protein, undergoes a sharp and transient activation in macrophages when phagocytosis was initiated via receptors for the Fc portion of immunoglobulins (FcRs). A dominant-negative mutant of ARF6 (T27N mutation) dramatically affected FcR-mediated phagocytosis. Expression of ARF6-T27N lead to a reduction in the focal delivery of vesicle-associated membrane protein 3+ endosomal recycling membranes at phagocytosis sites, whereas actin polymerization was unimpaired. This resulted in an early blockade in pseudopod extension and accumulation of intracellular vesicles, as observed by electron microscopy. We conclude that ARF6 is a major regulator of membrane recycling during phagocytosis

Heart ablation using a planar rectangular high intensity ultrasound transducer and MRI guidance

The aim of this study was to evaluate a flat rectangular (3 × 10 mm2) MRI compatible transducer operating at 5 MHz. The main task was to explore the feasibility of creating deep lesions in heart at a depth of at least 15 mm. The size of thermal necrosis in heart tissue was estimated as a function of power and time using a simulation model. The system was then tested in an excised lamb heart. In this study, we were able to create lesions of 15 mm deep with acoustic power of 6 W for an exposure of approximately 1 min. The contrast to noise ratio (CNR) between lesion and heart tissue was evaluated using fast spin echo (FSE). The CNR value was approximately 22 using T1 W FSE. Maximum CNR was achieved with repetition time (TR) between 300 and 800 ms. Using T2W FSE, the corresponding CNR was approximately 13 for the 14 in vivo experiments. The average lesion depth was 11.93 mm with a standard deviation of 0.62 mm. In vivo irradiation conditions were 6 W for 60 s. The size of the lesion in the other two dimensions was close to 3 × 10 mm2 (size of the transducer element)

A Role for Mammalian Diaphanous-Related Formins in Complement Receptor (CR3)-Mediated Phagocytosis in Macrophages

SummaryMacrophages, dendritic cells, and neutrophils use phagocytosis to capture and clear off invading pathogens. The process is triggered by the interaction of ligands on the pathogens’ surface with specific phagocytic receptors, including immunoglobulin (FcR) and complement C3bi (CR3) receptors (integrin αMβ2, Mac1) [1]. Localized actin-filament assembly that acts as the driving force for particle engulfment is controlled by Rho-family small GTPases [2, 3]. RhoA regulates CR3-mediated phagocytosis through a mechanism that is still unclear [4–6]. Mammalian Diaphanous-related (mDia) formins participate in the generation of a diverse set of actin-remodeling events downstream of RhoA [7], and mDia1 is recruited around fibronectin-coated beads in a RhoA-dependent manner in fibroblasts [8]. Here, we set out to examine whether mDia proteins are involved in CR3-mediated phagocytosis in macrophages. We show that the RhoA effector mDia1 is recruited early during CR3-mediated phagocytosis and colocalizes with polymerized actin in the phagocytic cup. Interfering with mDia activity inhibits CR3-mediated phagocytosis while having no effect on FcR-mediated phagocytosis. These results indicate a new function for mDia proteins in the regulation of actin polymerization during CR3-mediated phagocytosis

MT1-MMP directs force-producing proteolytic contacts that drive tumor cell invasion

International audienceUnraveling the mechanisms that govern the formation and function of invadopodia is essential towards the prevention of cancer spread. Here, we characterize the ultrastructural organization, dynamics and mechanical properties of collagenotytic invadopodia forming at the interface between breast cancer cells and a physiologic fibrillary type I collagen matrix. Our study highlights an uncovered role for MT1-MMP in directing invadopodia assembly independent of its proteolytic activity. Electron microscopy analysis reveals a polymerized Arp2/3 actin network at the concave side of the curved invadopodia in association with the collagen fibers. Actin polymerization is shown to produce pushing forces that repel the confining matrix fibers, and requires MT1-MMP matrix-degradative activity to widen the matrix pores and generate the invasive pathway. A theoretical model is proposed whereby pushing forces result from actin assembly and frictional forces in the actin meshwork due to the curved geometry of the matrix fibers that counterbalance resisting forces by the collagen fibers

Rab17, a novel small GTPase, is specific for epithelial cells and is induced during cell polarization

The rab subfamily of small GTPases has been demonstrated to play an important role in the regulation of membrane traffic in eukaryotic cells. Compared with nonpolarized cells, epithelial cells have distinct apical and basolateral transport pathways which need to be separately regulated. This raises the question whether epithelial cells require specific rab proteins. However, all rab proteins identified so far were found to be equally expressed in polarized and nonpolarized cells. Here we report the identification of rab17, the first epithelial cell-specific small GTPase. Northern blot analysis on various mouse organs revealed that the rab17 mRNA is present in kidney, liver, and intestine but not in organs lacking epithelial cells nor in fibroblasts. To determine whether rab17 is specific for epithelial cells we studied its expression in the developing kidney. We found that rab17 is absent from the mesenchymal precursors but is induced upon their differentiation into epithelial cells. In situ hybridization studies on the embryonic kidney and intestine revealed that rab17 is restricted to epithelial cells. By immunofluorescence and immunoelectron microscopy on kidney sections, rab17 was localized to the basolateral plasma membrane and to apical tubules. Rab proteins associated with two distinct compartments have been found to regulate transport between them. Therefore, our data suggest that rab17 might be involved in transcellular transport