Collective Cell Movement Research Articles

Supracellular cable drives collective cell movement

Neurodevelopment![Figure][1] Supracellular organization during collective migration CREDIT: SHELLARD ET AL. Neural crest cells migrate far and wide through a vertebrate embryo during development. Shellard et al. used Xenopus and zebrafish embryos to study how these clumps of mesenchymal cells migrate (see the Perspective by Adameyko). Movement was powered by a supracellular actomyosin cable that contracted around the rear of the clump. Similar supracellular contractility at the front was inhibited by a chemotactic signal. The imbalance in forces caused cells to rearrange so that the whole clump would be propelled forward. Science , this issue p. [339][2]; see also p. [290][3] [1]: pending:yes [2]: /lookup/doi/10.1126/science.aau3301 [3]: /lookup/doi/10.1126/science.aav3376

Local and global dynamics in collective movements of embryonic cells

Several important morphogenetic processes belong to the category of collective cell movements (CCM), by which we mean coordinated rearrangements of many neighboring cells. The causes of the dynamic order established during CCM are still unclear. We performed statistical studies of rates and angular orientations of cell rearrangements in two kinds of embryonic tissues, which we categorized as “committed” (in the sense of being capable of autonomous CCM) as opposed to “naïve” tissues, which are those that require external forces in order to exhibit full scale CCM. In addition, we distinguished two types of cell rearrangements: first, those in which mutual cell-cell shifts characterizing the local dynamics (LD); and, second, those which moved in reference to common external coordinates (global dynamics, GD). We observed that in most cases LD rates deviated from normal distributions and do so by creating excesses of extensively converging and moderately diverging cells. In contrast, GD was characterized by nearly random behavior of slowly moving cells, combined with increased angular focusing of the fast cells trajectories as well as bimodal distribution of cell rates. When committed tissues were opposed by external mechanical forces, then they tended to preserve the inherent CCM patterns. On the other hand, the naïve ones reacted by creating two orthogonal cells flows, one of these coinciding with the force direction. We consider CCM as a self-organizing process based on feedbacks between converging and diverging cell shifts, which is able to focus the trajectories imposed by external forces.

Cooperation of dual modes of cell motility promotes epithelial stress relaxation to accelerate wound healing.

Collective cell migration in cohesive units is vital for tissue morphogenesis, wound repair, and immune response. While the fundamental driving forces for collective cell motion stem from contractile and protrusive activities of individual cells, it remains unknown how their balance is optimized to maintain tissue cohesiveness and the fluidity for motion. Here we present a cell-based computational model for collective cell migration during wound healing that incorporates mechanochemical coupling of cell motion and adhesion kinetics with stochastic transformation of active motility forces. We show that a balance of protrusive motility and actomyosin contractility is optimized for accelerating the rate of wound repair, which is robust to variations in cell and substrate mechanical properties. This balance underlies rapid collective cell motion during wound healing, resulting from a tradeoff between tension mediated collective cell guidance and active stress relaxation in the tissue.

Fluid model of epithelial morphogenesis: Oscillations and structuring

We have revised a mathematical model of epithelial morphogenesis by Belintsev et al. (1987) (BB model) taking into account the oscillatory nature of morphogenesis and stability analysis (Cherdantsev, 2014). Following the BB model in considering the feedback control of cell shape changes by mechanical forces, we modify it to represent epithelial surface movements observed in different types of Metazoan gastrulation. Basing on these observations, we argue that the epithelial surface movement is that of an incompressible fluid supplemented by a positive feedback between the movement and spreading of the surface flow. Dipole interactions between sources and sinks of surface energy provide a single mechanism both of short-ranged and long-ranged regulation of collective cell and surface movements whose basic variables are the space averaged epithelial surface curvature and lateral pressure within the epithelial surface flux negatively related to its velocity. The short-ranged activation means a movement of the surface up to the lateral pressure gradient under non-linear feedback control of the surface flexure. The break of this feedback with equalization of the surface curvature is sufficient for a self-restriction of the movement spreading. Owing to bistable interdependence between the lateral pressure and epithelial surface curvature, we get a generic oscillatory contour in which the same region oscillates being alternately a sink and source of the surface flow. The opposite phase oscillations of the lateral pressure and curvature allow for both directional propulsion of the surface through the same region and spatial differentiation based on parametric differences between the large-scaled regions that correspond to sources and sinks of the surface.

Novel role for Grainy head in the regulation of cytoskeletal and junctional dynamics during epithelial repair.

Tissue repair is critical for the maintenance of epithelial integrity and permeability. Simple epithelial repair relies on a combination of collective cell movements and the action of a contractile actomyosin cable at the wound edge that together promote the fast and efficient closure of tissue discontinuities. The Grainy head family of transcription factors (Grh in flies; GRHL1-GRHL3 in mammals) are essential proteins that have been implicated both in the development and repair of epithelia. However, the genes and the molecular mechanisms that it controls remain poorly understood. Here, we show that Grh knockdown disrupts actomyosin dynamics upon injury of the Drosophila pupa epithelial tissue. This leads to the formation of an ectopic actomyosin cable away from the wound edge and impaired wound closure. We also uncovered that E-Cadherin is downregulated in the Grh-depleted tissue around the wound, likely as a consequence of Dorsal (an NF-κB protein) misregulation, which also affects actomyosin cable formation. Our work highlights the importance of Grh as a stress response factor and its central role in the maintenance of epithelial characteristics necessary for tissue repair through regulating cytoskeleton and E-Cadherin dynamics.

Friction-limited cell motility in confluent monolayer tissue

Mechanical forces are important factors in the development, coordination and collective motion of cells. Based on a continuum-scale model, we consider the influence of substrate friction on cell motility in confluent living tissue. We test our model on the experimental data of endothelial and cancer cells. In contrast to the commonly used drag friction, we find that solid friction best captures the cell speed distribution. From our model, we quantify a number of measurable physical tissue parameters, such as the ratio between the viscosity and substrate friction.

Epithelial vertex models with active biochemical regulation of contractility can explain organized collective cell motility.

Collective motions of groups of cells are observed in many biological settings such as embryo development, tissue formation, and cancer metastasis. To effectively model collective cell movement, it is important to incorporate cell specific features such as cell size, cell shape, and cell mechanics, as well as active behavior of cells such as protrusion and force generation, contractile forces, and active biochemical signaling mechanisms that regulate cell behavior. In this paper, we develop a comprehensive model of collective cell migration in confluent epithelia based on the vertex modeling approach. We develop a method to compute cell-cell viscous friction based on the vertex model and incorporate RhoGTPase regulation of cortical myosin contraction. Global features of collective cell migration are examined by computing the spatial velocity correlation function. As active cell force parameters are varied, we found rich dynamical behavior. Furthermore, we find that cells exhibit nonlinear phenomena such as contractile waves and vortex formation. Together our work highlights the importance of active behavior of cells in generating collective cell movement. The vertex modeling approach is an efficient and versatile approach to rigorously examine cell motion in the epithelium.

Three Dimensional Mixed-Cell Spheroids Mimic Stroma-Mediated Chemoresistance and Invasive Migration in hepatocellular carcinoma

Interactions between cancer cells and cancer-associated fibroblasts (CAFs) within the tumor microenvironment (TME) play an important role in promoting the profibrotic microenvironment and epithelial-mesenchymal transition (EMT), resulting in tumor progression and drug resistance in hepatocellular carcinoma (HCC). In the present study, we developed a mixed-cell spheroid model using Huh-7 HCC cells and LX-2 stellate cells to simulate the in vivo tumor environment with respect to tumor-CAF interactions. Spheroids were cultured from cancer cells alone (monospheroids) or as a mixture (mixed-cell spheroids) in ultra-low-attachment plates. Compact, well-mixed, and stroma-rich mixed-cell spheroids were successfully established with heterotypic cell-cell contacts shown by the presence of gap junctions and desmosomes. Mixed-cell spheroids showed enhanced expression of collagen type-I (Col‐I) and pro‐fibrotic factors such as, transforming growth factor beta1 (TGF-β1), and connective tissue growth factor (CTGF) compared to the levels expressed in mono-spheroids. The EMT phenotype was evident in mixed-cell spheroids as shown by the altered expression of E-cadherin and vimentin. Differential drug sensitivity was observed in mixed-cell spheroids, and only sorafenib and oxaliplatin showed dose-dependent antiproliferative effects. Simultaneous treatment with TGF-β inhibitors further improved sorafenib efficacy in the mixed-cell spheroids, indicating the involvement of TGF-β in the mechanism of sorafenib resistance. In 3D matrix invasion assay, mixed-cell spheroids exhibited fibroblast-led collective cell movement. Overall, our results provide evidence that mixed-cell spheroids formed with Huh-7 and LX-2 cells well represent HCC tumors and their TME in vivo and hence are useful in studying tumor-stroma interactions as mechanisms associated with drug resistance and increased cell motility.

An agent-based and FE approach to simulate cell jamming and collective motion in epithelial layers

The collective cell motion in epithelial layers is still poorly understood, despite this phenomenon being fundamental to explain several biological processes. Indeed, it has been experimentally observed that epithelial cells can behave either as a fluid or as a solid in the tissue. The transition between both states is related to cell–cell adhesions and cell morphology. In fact, cell motility can be limited by the interaction with its neighboring cells. Moreover, cells can even enter in a frozen state, and in that case, the system behaves as a solid. However, this state is reversible under certain circumstances, and cells may return to a fluidized state. This phenomenon is known as cell jamming. Here, we propose a hybrid approach that couples a discrete agent-based model and a continuum finite element-based model to simulate cell dynamics and cell jamming in epithelial monolayers. Our hybrid approach is able to simulate cell motion individually, but it also reproduces the mechanical properties at tissue level that emerge from cell–cell interactions. This study helps to understand how cell–cell interactions regulate the cell jamming phenomenon and provides a deeper insight into the role of the passive mechanics in collective cell motion.

α-Catenin Controls the Anisotropy of Force Distribution at Cell-Cell Junctions during Collective Cell Migration

Adherens junctions (AJs) control epithelial cell behavior, such as collective movement and morphological changes, during development and in disease. However, the molecular mechanism of AJ remodeling remains incompletely understood. Here, we report that the conformational activation of α-catenin is the key event in the dynamic regulation of AJ remodeling. α-catenin activates RhoA to increase actomyosin contractility at cell-cell junctions. This leads to the stabilization of activated α-catenin, in part through the recruitment of the actin-binding proteins, vinculin and afadin. In this way, α-catenin regulates force sensing, as well as force transmission, through a Rho-mediated feedback mechanism. We further show that this is important for stable directional alignment of multiple cells during collective cell movement by both experimental observation and mathematical modeling. Taken together, our findings demonstrate that α-catenin controls the establishment of anisotropic force distribution at cell junctions to enable cooperative movement of the epithelial cell sheet.

A DISCRETE MATHEMATICAL MODEL FOR SINGLE AND COLLECTIVE MOVEMENT IN AMOEBOID CELLS

In this paper, we develop a new discrete mathematical model for individual and collective cell motility. We introduce a mechanical model for the movement of a cell on a two-dimensional rigid surface to describe and investigate the cell–cell and cell–substrate interactions. The cell cytoskeleton is modeled as a series of springs and dashpots connected in parallel. The cell–substrate attachments and the cell protrusions are also included. In particular, this model is used to describe the directed movement of endothelial cells on a Matrigel plate. We compare the results from our model with experimental data. We show that cell density and substrate rigidity play an important role in network formation.

Deep reinforcement learning of cell movement in the early stage of C.elegans embryogenesis.

Cell movement in the early phase of Caenorhabditis elegans development is regulated by a highly complex process in which a set of rules and connections are formulated at distinct scales. Previous efforts have demonstrated that agent-based, multi-scale modeling systems can integrate physical and biological rules and provide new avenues to study developmental systems. However, the application of these systems to model cell movement is still challenging and requires a comprehensive understanding of regulatory networks at the right scales. Recent developments in deep learning and reinforcement learning provide an unprecedented opportunity to explore cell movement using 3D time-lapse microscopy images. We present a deep reinforcement learning approach within an agent-based modeling system to characterize cell movement in the embryonic development of C.elegans. Our modeling system captures the complexity of cell movement patterns in the embryo and overcomes the local optimization problem encountered by traditional rule-based, agent-based modeling that uses greedy algorithms. We tested our model with two real developmental processes: the anterior movement of the Cpaaa cell via intercalation and the rearrangement of the superficial left-right asymmetry. In the first case, the model results suggested that Cpaaa's intercalation is an active directional cell movement caused by the continuous effects from a longer distance (farther than the length of two adjacent cells), as opposed to a passive movement caused by neighbor cell movements. In the second case, a leader-follower mechanism well explained the collective cell movement pattern in the asymmetry rearrangement. These results showed that our approach to introduce deep reinforcement learning into agent-based modeling can test regulatory mechanisms by exploring cell migration paths in a reverse engineering perspective. This model opens new doors to explore the large datasets generated by live imaging. Source code is available at https://github.com/zwang84/drl4cellmovement. Supplementary data are available at Bioinformatics online.

Dynamic force patterns promote collective cell movements during embryonic wound repair

Embryonic wounds heal rapidly, in a process driven by coordinated cell movements. Polarization of actin and the molecular motor non-muscle myosin II in the cells adjacent to the wound results in the formation of a supracellular cable around the wound that drives repair. In Drosophila embryos, the distribution of actin around wounds is heterogeneous, with regions of high and low actin density, and actin heterogeneity is necessary for rapid repair. Here, we demonstrate that actin and myosin display stochastic patterns around embryonic wounds, and that contractile forces around wounds are heterogeneous. Mathematical modelling suggests that actomyosin heterogeneity favours wound closure if myosin is regulated by tension and strain, a hypothesis that we validate experimentally. We show that inhibition of stretch-activated ion channels disrupts myosin dynamics and tissue repair. Together, our results indicate that staggered contractile events, mechanical signals and force-regulated myosin dynamics coordinate cell behaviours to drive efficient wound closure. The motor proteins and contractile forces involved in wound closure are both shown to be heterogeneously distributed around a wound. Theory suggests that this heterogeneity speeds up wound closure, as long as the proteins are mechanically regulated.

Statistical modelling of cell movement

Collective cell movement affects vital biological processes in the human organism such as wound healing, immune response, and cancer metastasis. A better understanding of the mechanisms driving cell movement is then essential for the advancement of medical treatments. In this paper, we demonstrate how the unscented Kalman filter, a technique used extensively in engineering in the context of filtering, can be applied to estimate random or directed cell movement. Our proposed model, formulated using stochastic differential equations, is fitted on data describing the movement of Dictyostelium cells, an amoeba routinely used as a proxy for eukaryotic cell movement.

Cdc42 controls primary mesenchyme cell morphogenesis in the sea urchin embryo

In the sea urchin embryo, gastrulation is characterized by the ingression and directed cell migration of primary mesenchyme cells (PMCs), as well as the primary invagination and convergent extension of the endomesoderm. Like all cell shape changes, individual and collective cell motility is orchestrated by Rho family GTPases and their modulation of the actomyosin cytoskeleton. And while endomesoderm specification has been intensively studied in echinoids, much less is known about the proximate regulators driving cell motility. Toward these ends, we employed anti-sense morpholinos, mutant alleles and pharmacological inhibitors to assess the role of Cdc42 during sea urchin gastrulation. While inhibition of Cdc42 expression or activity had only mild effects on PMC ingression, PMC migration, alignment and skeletogenesis were disrupted in the absence of Cdc42, as well as elongation of the archenteron. PMC migration and patterning of the larval skeleton relies on the extension of filopodia, and Cdc42 was required for filopodia in vivo as well as in cultured PMCs. Lastly, filopodial extension required both Arp2/3 and formin actin-nucleating factors, supporting models of filopodial nucleation observed in other systems. Together, these results suggest that Cdc42 plays essential roles during PMC cell motility and organogenesis.

E-cadherin mediated lateral interactions between neighbor cells necessary for collective migration

Collective cell movement is critical in pathological processes such as wound healing and cancer invasion. It entails complex interactions between adjacent cells and between cells-extracellular matrices. Most studies measure the migration patterns and force propagation by placing cells on flat, patterned substrates. The cooperative behavior resulting from cell-cell interactions is not well understood. We have developed a multi-channel microfluidic device that has junctional protein E-cadherin coated onto the sidewalls of the channels that enables the cells’ lateral interactions with their neighbors to be studied. Our study reveals that epithelial cells rely on lateral E-cadherin-based adhesions to maintain the cohesion of the group. Cells move faster in narrower channels, but the average velocity along the channels is reduced in E-cadherin coated channels versus non-adhesive channels. We have directly measured the forces in the cross-linking protein, alpha-actinin, using FRET sensors during cell migration, and found that higher tension exists at the cell edges adjacent to the walls coated with E-cadherin, the implication being E-cadherin transmits the shear forces but does not provide a driving force for this migration.

Dia1-dependent adhesions are required by epithelial tissues to initiate invasion.

Developing tissues change shape and tumors initiate spreading through collective cell motility. Conserved mechanisms by which tissues initiate motility into their surroundings are not known. We investigated cytoskeletal regulators during collective invasion by mouse tumor organoids and epithelial Madin-Darby canine kidney (MDCK) acini undergoing branching morphogenesis in collagen. Use of the broad-spectrum formin inhibitor SMIFH2 prevented the formation of migrating cell fronts in both cell types. Focusing on the role of the formin Dia1 in branching morphogenesis, we found that its depletion in MDCK cells does not alter planar cell motility either within the acinus or in two-dimensional scattering assays. However, Dia1 was required to stabilize protrusions extending into the collagen matrix. Live imaging of actin, myosin, and collagen in control acini revealed adhesions that deformed individual collagen fibrils and generated large traction forces, whereas Dia1-depleted acini exhibited unstable adhesions with minimal collagen deformation and lower force generation. This work identifies Dia1 as an essential regulator of tissue shape changes through its role in stabilizing focal adhesions.

Collective Cell Migration in Embryogenesis Follows the Laws of Wetting

Collective cell migration is a fundamental process during embryogenesis and its initial occurrence, called epiboly, is an excellent in vivo model to study the physical processes involved in collective cell movements that are key to understanding organ formation, cancer invasion, and wound healing. In zebrafish, epiboly starts with a cluster of cells at one pole of the spherical embryo. These cells are actively spreading in a continuous movement toward its other pole until they fully cover the yolk. Inspired by the physics of wetting, we determine the contact angle between the cells and the yolk during epiboly. By choosing a wetting approach, the relevant scale for this investigation is the tissue level, which is in contrast to other recent work. Similar to the case of a liquid drop on a surface, one observes three interfaces that carry mechanical tension. Assuming that interfacial force balance holds during the quasi-static spreading process, we employ the physics of wetting to predict the temporal change of the contact angle. Although the experimental values vary dramatically, the model allows us to rescale all measured contact-angle dynamics onto a single master curve explaining the collective cell movement. Thus, we describe the fundamental and complex developmental mechanism at the onset of embryogenesis by only three main parameters: the offset tension strength, α, that gives the strength of interfacial tension compared to other force-generating mechanisms; the tension ratio, δ, between the different interfaces; and the rate of tension variation, λ, which determines the timescale of the whole process.

FGF controls epithelial-mesenchymal transitions during gastrulation by regulating cell division and apicobasal polarity.

To support tissue and organ development, cells transition between epithelial and mesenchymal states. Here, we have investigated how mesoderm cells change state in Drosophila embryos and whether fibroblast growth factor (FGF) signaling plays a role. During gastrulation, presumptive mesoderm cells invaginate, undergo an epithelial-to-mesenchymal state transition (EMT) and migrate upon the ectoderm. Our data show that EMT is a prolonged process in which adherens junctions progressively decrease in number throughout the migration of mesoderm cells. FGF influences adherens junction number and promotes mesoderm cell division, which we propose decreases cell-cell attachments to support slow EMT while retaining collective cell movement. We also found that, at the completion of migration, cells form a monolayer and undergo a reverse mesenchymal-to-epithelial transition (MET). FGF activity leads to accumulation of β-integrin Myospheroid basally and cell polarity factor Bazooka apically within mesoderm cells, thereby reestablishing apicobasal cell polarity in an epithelialized state in which cells express both E-Cadherin and N-Cadherin. In summary, FGF plays a dynamic role in supporting mesoderm cell development to ensure collective mesoderm cell movements, as well as proper differentiation of mesoderm cell types.

Force-dependent binding of vinculin to α-catenin regulates cell-cell contact stability and collective cell behavior.

The shaping of a multicellular body and repair of adult tissues require fine--tuning of cell adhesion, cell mechanics, and intercellular transmission of mechanical load. Adherens junctions (AJs) are the major intercellular junctions by which cells sense and exert mechanical force on each other. However, how AJs adapt to mechanical stress and how this adaptation contributes to cell-cell cohesion and eventually to tissue-scale dynamics and mechanics remains largely unknown. Here, by analyzing the tension-dependent recruitment of vinculin, α-catenin, and F-actin as a function of stiffness, as well as the dynamics of GFP-tagged wild-type and mutated α-catenins, altered for their binding capability to vinculin, we demonstrate that the force-dependent binding of vinculin stabilizes α-catenin and is responsible for AJ adaptation to force. Challenging cadherin complexes mechanical coupling with magnetic tweezers, and cell-cell cohesion during collective cell movements, further highlight that tension-dependent adaptation of AJs regulates cell-cell contact dynamics and coordinated collective cell migration. Altogether, these data demonstrate that the force-dependent α-catenin/vinculin interaction, manipulated here by mutagenesis and mechanical control, is a core regulator of AJ mechanics and long-range cell-cell interactions.