Angélique Stéphanou

The motility of normal and cancer cells in response to the combined influence of the substrate rigidity and anisotropic microstructure

Cell adhesion and migration are strongly influenced by extracellular matrix (ECM) architecture and rigidity, but little is known about the concomitant influence of such environmental signals to cell responses, especially when considering cells of similar origin and morphology, but exhibiting a normal or cancerous phenotype. Using micropatterned polydimethylsiloxane substrates (PDMS) with tunable stiffness (500 kPa, 750 kPa, 2000 kPa) and topography (lines, pillars or unpatterned), we systematically analyse the differential response of normal (3T3) and cancer (SaI/N) fibroblastic cells. Our results demonstrate that both cells exhibit differential morphology and motility responses to changes in substrate rigidity and microtopography. 3T3 polarisation and spreading are influenced by substrate microtopography and rigidity. The cells exhibit a persistent type of migration, which depends on the substrate anisotropy. In contrast, the dynamic of SaI/N spreading is strongly modified by the substrate topography but not by substrate rigidity. SaI/N morphology and migration seem to escape from extracellular cues: the cells exhibit uncorrelated migration trajectories and a large dispersion of their migration speed, which increases with substrate rigidity.

Mathematical modelling of the influence of blood rheological properties upon adaptative tumour-induced angiogenesis

In this paper, we present a theoretical investigation of the influence of blood flow through a tumour-induced capillary network, whereby the vascular architecture adapts as it grows to the associated haemodynamic forces resulting in what we describe as adaptive tumour-induced angiogenesis (ATIA). The network is generated in response to tumour angiogenic factors (TAFs), which are released from hypoxic cells within a solid tumour. We first describe a refined model for tumour-induced angiogenesis, which aims to describe the capillary growth process at the cellular level by explicitly taking into account the effects of matrix degrading enzymes and the local properties of the host tissue during endothelial cell migration. We then incorporate blood rheological properties into the formulation and investigate the influence of wall shear stress induced by the blood flow during dynamic vascular growth. We then go on to examine a number of feedback mechanisms affecting vascular resistance and network architecture. The mechanisms considered include those proposed by Pries and co-workers [A.R. Pries, T.W. Secomb, P. Gaehtgens, Structural adaptation and stability of microvascular networks: theory and simulation, Am. J. Physiol. Heart Circ. Physiol. 44 (1998) H349-H360; A.R. Pries, B. Reglin, T.W. Secomb, Structural adaptation of microvascular networks: functional roles of adaptative responses, Am. J. Physiol. Heart Circ. Physiol. 281 (2001) H1015-H1025; A.R. Pries, B. Reglin, T.W. Secomb, Structural adaptation of microvascular networks: roles of the pressure response, Hypertension 38 (2001) 1476-1479] and both haemodynamic (non-linear viscosity) and metabolic constraints are taken into account. Subsequent simulations of chemotherapeutic drug perfusion through the system show that vascular adaptation leads to a significant benefit in treatment delivery to the tumour. The results clearly demonstrate that the combined effects of network architecture and vessel compliance should be included in future models of angiogenesis if therapy protocols and treatment efficacy are to be adequately assessed.

Spatiotemporal dynamics of actin-rich adhesion microdomains: influence of substrate flexibility

In this study we analyse the formation and dynamics of specific actin-rich structures called podosomes. Podosomes are very dynamic punctual adhesion sites tightly linked to the actin cytoskeleton. Mechanical properties of substrates are emerging as important physical modulators of anchorage-dependent processes involved in the cellular response. We investigate the influence of substrate flexibility on the dynamic properties of podosomes. We used mouse NIH-3T3 fibroblasts, transfected with GFP-actin and cultured on polyacrylamide collagen-coated substrates of varying stiffness. Static and dynamic features of cell morphologies associated with an optical flow analysis of the dynamics of podosomes revealed that: (1) they have constant structural properties, i.e. their shape factor and width do not change with the substrate flexibility; (2) the lifespan of podosomes and mean minimum distance between them depend on the substrate flexibility; (3) there is a variation in the displacement speed of the rosette of podosomes. Moreover, the rosettes sometimes appear as periodically emergent F-actin structures, which suggests that a two-level self-organisation process may drive first, the formation of clusters of podosomes and second, the organisation of these clusters into oscillating rings. Such dynamic features give new perspectives regarding the potential function of podosomes as mechanosensory structures.

A computational model of cell migration coupling the growth of focal adhesions with oscillatory cell protrusions

Cell migration is a highly integrated process where actin turnover, actomyosin contractility, and adhesion dynamics are all closely linked. In this paper, we propose a computational model investigating the coupling of these fundamental processes within the context of spontaneous (i.e. unstimulated) cell migration. In the unstimulated cell, membrane oscillations originating from the interaction between passive hydrostatic pressure and contractility are sufficient to lead to the formation of adhesion spots. Cell contractility then leads to the maturation of these adhesion spots into focal adhesions. Due to active actin polymerization, which reinforces protrusion at the leading edge, the traction force required for cell translocation can be generated. Computational simulations first show that the model hypotheses allow one to reproduce the main features of fibroblast cell migration and established results on the biphasic aspect of the cell speed as a function of adhesion strength. The model also demonstrates that certain temporal parameters, such as the adhesion proteins recycling time and adhesion lifetimes, influence cell motion patterns, particularly cell speed and persistence of the direction of migration. This study provides some elements, which allow a better understanding of spontaneous cell migration and enables a first glance at how an individual cell would potentially react once exposed to a stimulus.

Cytomechanics of cell deformations and migration: from models to experiments

A cytomechanical model bas been proposed to analyse cell-cell interactions and cell migration through chemotaxis. We consider as the leading assumption that the cell cortical tension is locally modified by the protrusive activity of neighbour cells and binding of chemoattractant molecules to membrane receptors respectively. The model derives from the one initially proposed by Alt and Tranquillo (1995), which successfully describes experimentally observed cyclic autonomous cell shape changes. It is based on force balance equations coupling intracellular hydrostatic pressure and cell cortex contraction. Considering the protrusive dynamics of L929 fibroblats observed by videomicroscopy, we simulated the influence of neighbouring protrusions on a cell spontaneous pulsating behaviour. We further investigated the role of an extracellular gradient as another kind of external stimulus. The model illustrates how binding of chemoattractant molecules can induce a cell morphological instability that, above an intracellular stress threshold, will break the cell-substratum attachment. As a result, realistic cell chemotaxis can be simulated.

On the importance of the submicrovascular network in a computational model of tumour growth

A computational model is potentially a powerful tool to apprehend complex phenomena like solid tumour growth and to predict the outcome of therapies. To that end, the confrontation of the model with experiments is essential to validate this tool. In this study, we develop a computational model specifically dedicated to the interpretation of tumour growth as observed in a mouse model with a dorsal skinfold chamber. Observation of the skin vasculature at the dorsal window scale shows a sparse network of a few main vessels of several hundreds micrometers in diameter. However observation at a smaller scale reveals the presence of a dense and regular interconnected network of capillaries about ten times smaller. We conveniently designate this structure as the submicrovascular network (SMVN).(1) The question that we wish to answer concerns the necessity of explicitly taking into account the SMVN in the computational model to describe the tumour evolution observed in the dorsal chamber. For that, simulations of tumour growth realised with and without the SMVN are compared and lead to two distinct scenarios. Parameters that are known to strongly influence the tumour evolution are then tested in the two cases to determine to which extent those parameters can be used to compensate the observed differences between these scenarios. Explicit modelling of the smallest vessels appears mandatory although not necessarily under the form of a regular grid. A compromise between the two investigated cases can thus be reached.

How tumour-induced vascular changes alter angiogenesis: Insights from a computational model

A computational model was developed to describe experimentally observed vascular changes induced by the introduction of a tumour on a mouse equipped with a dorsal skinfold chamber. The vascular structure of the host tissue was segmented from in vivo images and transposed into the computational framework. Simulations of tumour-induced vascular changes were performed and include the destabilizing effects of the growth factor VEGF on the integrity of the vessels walls. The integration of those effects, that include alteration of the vessel wall elasticity and wall breaching, were required to realistically reproduce the experimental observations. The model was then used to investigate the importance of the vascular changes for oxygen delivery and tumour development. To that end, we compared simulations obtained with a dynamic vasculature with those obtained with a static one. The results showed that the tumour growth was strongly impeded by the constant vascular changes. More precisely, it is the angiogenic process itself that was affected by vascular changes occurring in bigger upstream vessels and resulting in a less efficient angiogenic network for oxygen delivery. As a consequence, tumour cells are mostly kept in a non-proliferative hypoxic state. Tumour dormancy thus appears as one potential consequence of the intense vascular changes in the host tissue.

Towards the Design of a Patient-Specific Virtual Tumour

The design of a patient-specific virtual tumour is an important step towards Personalized Medicine. However this requires to capture the description of many key events of tumour development, including angiogenesis, matrix remodelling, hypoxia, and cell state heterogeneity that will all influence the tumour growth kinetics and degree of tumour invasiveness. To that end, an integrated hybrid and multiscale approach has been developed based on data acquired on a preclinical mouse model as a proof of concept. Fluorescence imaging is exploited to build case-specific virtual tumours. Numerical simulations show that the virtual tumour matches the characteristics and spatiotemporal evolution of its real counterpart. We achieved this by combining image analysis and physiological modelling to accurately described the evolution of different tumour cases over a month. The development of such models is essential since a dedicated virtual tumour would be the perfect tool to identify the optimum therapeutic strategies that would make Personalized Medicine truly reachable and achievable.

Role of Compartmentalization on HiF-1α Degradation Dynamics during Changing Oxygen Conditions: A Computational Approach

HiF-1α is the central protein driving the cellular response to hypoxia. Its accumulation in cancer cells is linked to the appearance of chemoresistant and aggressive tumor phenotypes. As a consequence, understanding the regulation of HiF-1α dynamics is a major issue to design new anti-cancer therapies. In this paper, we propose a model of the hypoxia pathway, involving HiF-1α and its inhibitor pVHL. Based on data from the literature, we made the hypothesis that the regulation of HiF-1α involves two compartments (nucleus and cytoplasm) and a constitutive shuttle of the pVHL protein between them. We first show that this model captures correctly the main features of HiF-1α dynamics, including the bi-exponential degradation profile in normoxia, the kinetics of induction in hypoxia, and the switch-like accumulation. Second, we simulated the effects of a hypoxia/reoxygenation event, and show that it generates a strong instability of HiF-1α. The protein concentration rapidly increases 3 hours after the reoxygenation, and exhibits an oscillating pattern. This effect vanishes if we do not consider compartmentalization of HiF-1α. This result can explain various counter-intuitive observations about the specific molecular and cellular response to the reoxygenation process. Third, we simulated the HiF-1α dynamics in the tumor case. We considered different types of mutations associated with tumorigenesis, and we compared their consequences on HiF-1α dynamics. Then, we tested different therapeutics strategies. We show that a therapeutic decrease of HiF-1α nuclear level is not always correlated with an attenuation of reoxygenation-induced instabilities. Thus, it appears that the design of anti-HiF-1α therapies have to take into account these two aspects to maximize their efficiency.

XXXIVth Seminar of the French-Speaking Society for Theoretical Biology: Saint-Flour (Cantal), France, 26–28 May, 2014

Historically, theoretical biology was built on mathematics in Europe. The FrenchSpeaking Society of Theoretical Biology (SFBT), strong with a long tradition in biomathematics, federates theoretical biology in French-Speaking countries and France in particular. For several years now, with the increase of computational means and the development of research in the domain of complex systems, the framework of theoretical biology evolved towards more computational approaches. The Spring School 2012 of the SFBT reflected this trend by addressing the theme of numerical experimentations and hybrid approaches which associate continuous and discrete models. On this 2014 edition of the seminar of the society, we wished to go one step further by promoting discussions on the subject of Artificial Life. Both Theoretical Biology and Artificial Life aim to understand the Living. The difference is essentially a semantical one and is often linked to the scale at which the problem is addressed. In theoretical biology, we ‘‘make models’’, often along a topdown approach on the basis of mathematical equations. In artificial life, we ‘‘simulate virtual words’’ along a reciprocal bottom-up approach based on numerical entities. As different as they may appear and as different as the associated communities may be, their aim remains the same: deciphering the mechanisms of the Living and forecasting its evolution. The two approaches converge in the sense that each provides some elements of understanding. The first paper by Atangana ‘‘Modeling the enzyme kinetic reaction’’ is such a contribution for the understanding of enzymatic mechanisms. In the other hand, the following paper by Bedessem