Probing mechanical properties to study cancer cell migration

To best comprehend cellular behaviour and how it determines cell migration in metastatic cancer, the research described here has focused on cell mechanics. The signalling pathway involving Rho-associated kinase (ROCK) has emerged as being the main regulator for the cellular cytoskeleton and actomyosin contractility that play key roles in metastatic cancer formation. In this thesis, an examination is made of how the cellular properties intertwine as ROCK is overexpressed. In research towards being able to measure and describe the viscoelastic properties of a cell that are associated with cell mechanics, over a wide range of timescales, a novel AFM force indentation data analysis method was applied. In particular, as part of this study, pancreatic ductal adenocarcinoma (PDAC) cells were overexpressed with ROCK, and the influence of ROCK activity on cell’s elastic and viscoelastic properties were quantified. It was found that when ROCK activity was overexpressed in cells, their elasticity decreased while their viscosity remained unchanged. These properties had a direct correlation with the activity of ADF/cofilin - the proteins downstream of ROCK. This meant that with overexpression, more stable actin bundles were present along with their inward stresses generated by the actomyosin contraction. This is consistent with an increased level of compressive forces within cells. Collective compressive forces between cell-cell are associated with the packing of cells that decreases cellular response. To further understand the role of ROCK activity in cancer invasion, a microfluidic device was created to mimic cell migration through tissue. The device consists of precisely defined microchannels with dimensions chosen to hinder and confine the cells in a manner similar to that found in a physiological environment. It was found that overexpressed ROCK1 cells in the confinement had notable decrease in cell size and motility. Along with this decrease in mechanical properties, observations also gave rise to questions about the connection between these properties that remain to be answered

Formation and evolution of globular clusters in a cosmological context

Stellar clusters are observed in a variety of galactic environments, from their current formation sites like the disks of the Antennae galaxies to the old globular cluster populations which in the Milky Way mostly reside in the halo. This suggests that the evolution of these puzzling objects may be linked to that of their host galaxy. This thesis explores the formation and evolution of stellar clusters in a cosmological context. For that, analytical models are developed to describe the role of the galactic environment in shaping their demographics. A suite of cosmological, hydrodynamical simulations of Milky Way-mass galaxies from the E-MOSAICS project are used to study when stellar clusters form and how they evolve over cosmic history. These simulations are also employed to estimate the contribution of stellar clusters to the build-up of stellar haloes. Finally, the EMP-Pathfinder simulations are presented, which represent the next generation of simulations of the co-formation and evolution of stellar clusters alongside their host galaxies in a cold, dense cosmic environment. The conclusion drawn from these studies is that stellar clusters are tightly linked to their host cosmic environments. This leads to exciting new future directions that are briefly discussed

Modélisation et simulation 3D de la morphogenèse

The embryo of the Drosophila Melanogaster undergoes a series of cell movements during its early development. Gastrulation is the process describing the segregation of the future internal tissues into the interior of the developing embryo. Gastrulation starts with the formation of the ventral furrow, a process commonly known as the ventral furrow invagination. During this process, the most ventrally located blastoderm cells flatten and progressively constrict their apical sides until they are wedge shaped. As a result of these cell-shape changes, the blastoderm epithelium first forms an indentation, the ventral furrow, which is then completely internalized. We focus on the study of the mechanisms that drive the invagination. The main questions that gave birth to this thesis are: “What is the role of the apical constriction of the ventral cells in the invagination?” and “Once the ventral cells are internalized, what is the mechanism that drives the ventral closure?” We attempt to answer to these two questions from a biomechanical point of view. For this purpose, a 3D mesh of the embryo of the Drosophila Melanogaster has been created. Based on this mesh, two “a minima” biomechanical models of the Drosophila embryo have been created, a physically based discrete model and a model based on the Finite Element Method. The results of the simulations in both models show that the geometry of the embryo plays a crucial role in the internalization of the ventral cells. The two models efficiently simulate the internalization of the ventral cells but are incapable of reproducing the ventral closure. We hypothesize that the ventral closure can be explained by the interplay of forces developed in the embryo once the internalized ventral cells undergo cell division. We propose an approach to divide elements in a Finite Element Mesh and we integrate it to the Finite Element Model of the Drosophila Melanogaster.L'embryon de la Drosophila Melanogaster subit une série des mouvements cellulaires pendant son développement. La gastrulation est le processus qui décrit la différentiation des futurs tissus à l'intérieur de l'embryon. La gastrulation commence par la formation du sillon ventral, un processus connu sous le nom de “Ventral Furrow Invagination”. Pendant ce processus, les cellules de la blastoderme positionnées dans la région ventrale de l'embryon, aplatissent et contractent leur surface apicale jusqu'à ce qu'elles deviennent prismatiques. Ce changement de forme cellulaire aboutit à un enfoncement au niveau de la région ventrale, le sillon ventral, qui est ensuite totalement intériorisé. Nous focalisons notre étude sur les mécanismes qui conduisent à l'invagination. Les questions principales auxquelles ce travail de thèse essaie de répondre sont: “Quel est le rôle de la contraction apicale des cellules ventrales dans l'invagination?” et “Quel est le mécanisme qui conduit à la clôture ventrale, une fois les cellules ventrales intériorisées?”. Nous essayons de répondre à ces questions d'un point de vue biomécanique. Dans ce but, un maillage 3D de l'embryon de la Drosophila Melanogaster a été créé. Basés sur ce maillage, deux modèles biomécaniques “a minima” de l'embryon de la Drosophila ont été créés: un modèle physique discret et un modèle basé sur la Méthode des Eléments Finis. Les résultats des simulations des deux modèles montrent que la géométrie joue un rôle décisif dans l'intériorisation des cellules ventrales. Les deux modèles ont permis de simuler l'intériorisation des cellules ventrales mais se trouvent incapables de simuler la clôture ventrale. Notre hypothèse est que la clôture ventrale peut s'expliquer par l'intéraction des forces développées à l'intérieur de l'embryon, une fois que les cellules ventrales commencent à proliférer. Nous proposons une méthode pour diviser des éléments dans un maillage d'éléments finis et ensuite nous expliquons l'intégration de cette méthode dans le modèle des Eléments Finis pour l'embryon de la Drosophila Melanogaster

Snail-like pattern generation with the Hénon family of maps

9 pages, 7 figures.-- PACS nrs.: 05.45.Ac, 47.52+j.-- Printed version published on Jun 2001.In this paper we show that in the Hénon family of maps x(n+1)=A+By(n)−x(n)^2, y(n+1)=x(n), for given parameter values, the orbit of an initial segment has a snail-like pattern that is related to the starfish-like pattern of the orbit of an initial point for the same parameter values. We study the parameter plane in the Hénon family of maps, searching for orbits with both starfish-like and snail-like patterns.This research was supported by Comunidad de Madrid and DGESIC, Spain, under grants 07/0044/1998 and TEL98-1020, respectively.Peer reviewe