Mechanochemical Feedback Research Articles

Sculpting with stem cells: how models of embryo development take shape.

ABSTRACTDuring embryogenesis, organisms acquire their shape given boundary conditions that impose geometrical, mechanical and biochemical constraints. A detailed integrative understanding how these morphogenetic information modules pattern and shape the mammalian embryo is still lacking, mostly owing to the inaccessibility of the embryo in vivo for direct observation and manipulation. These impediments are circumvented by the developmental engineering of embryo-like structures (stembryos) from pluripotent stem cells that are easy to access, track, manipulate and scale. Here, we explain how unlocking distinct levels of embryo-like architecture through controlled modulations of the cellular environment enables the identification of minimal sets of mechanical and biochemical inputs necessary to pattern and shape the mammalian embryo. We detail how this can be complemented with precise measurements and manipulations of tissue biochemistry, mechanics and geometry across spatial and temporal scales to provide insights into the mechanochemical feedback loops governing embryo morphogenesis. Finally, we discuss how, even in the absence of active manipulations, stembryos display intrinsic phenotypic variability that can be leveraged to define the constraints that ensure reproducible morphogenesis in vivo.

Mechanical oscillations orchestrate axial patterning through Wnt activation in Hydra.

Mechanical input shapes cell fate decisions during development and regeneration in many systems, yet the mechanisms of this cross-talk are often unclear. In regenerating Hydra tissue spheroids, periodic osmotically driven inflation and deflation cycles generate mechanical stimuli in the form of tissue stretching. Here, we demonstrate that tissue stretching during inflation is important for the appearance of the head organizer—a group of cells that secrete the Wnt3 ligand. Exploiting time series RNA expression profiles, we identify the up-regulation of Wnt signaling as a key readout of the mechanical input. In this system, the levels of Wnt3 expression correspond to the levels of stretching, and Wnt3 overexpression alone enables successful regeneration in the absence of mechanical stimulation. Our findings enable the incorporation of mechanical signals in the framework of Hydra patterning and highlight the broad significance of mechanochemical feedback loops for patterning epithelial lumens.

Multicellular mechanochemical hybrid cellular Potts model of tissue formation during epithelial‐mesenchymal transition

AbstractEpithelial‐mesenchymal transition (EMT) is the transdifferentiation of epithelial cells to a mesenchymal phenotype, in which cells lose epithelial‐like cell–cell adhesions and gain mesenchymal‐like enhanced contractility and mobility. EMT is crucial for tissue regeneration and is also implicated in pathological conditions, such as cancer metastasis. Prior work has shown that transforming growth factor‐1 (TGF‐1) is a potent inducer of this biological process. In this study, we develop a computational model coupling mechanical and biochemical signaling in a multicellular tissue undergoing EMT. Specifically, we utilize a recently developed formulation that integrates a multicellular cellular Potts model (CPM), a lattice‐based stochastic model governing cell movement; a first moment of area model, governing cellular traction and junctional forces; a finite element model, which defines extracellular matrix (ECM) substrate strains; an intracellular signaling TGF‐1‐mediated EMT model that governs cellular phenotype; and an extracellular signaling component governing ECM and TGF‐1 signaling. In this study, we modeled the spatial cellular patterns that occur in tissue and the ECM during EMT. Our model predicts that EMT often initially occurs at a tissue boundary due to mechanochemical coupling, which results in transdifferentiation to progress inwards toward the center. Variation in model parameters demonstrated conditions enhancing and suppressing EMT, especially to drive EMT in the absence of TGF‐1 and inhibit EMT in the presence of TGF‐1. Specifically, enhancing the mechanochemical feedback typically promoted EMT, whereas greater assembled ECM degradation suppressed EMT. Simulated scratch test experiments illustrate that ECM composition can impact closure directly through EMT signaling. In conclusion, we integrated mechanical, biochemical, and extracellular signaling networks in a novel hybrid computational model to reproduce tissue formation dynamics of EMT.

Dynamic mechanochemical feedback between curved membranes and BAR protein self-organization

In many physiological situations, BAR proteins reshape membranes with pre-existing curvature (templates), contributing to essential cellular processes. However, the mechanism and the biological implications of this reshaping process remain unclear. Here we show, both experimentally and through modelling, that BAR proteins reshape low curvature membrane templates through a mechanochemical phase transition. This phenomenon depends on initial template shape and involves the co-existence and progressive transition between distinct local states in terms of molecular organization (protein arrangement and density) and membrane shape (template size and spherical versus cylindrical curvature). Further, we demonstrate in cells that this phenomenon enables a mechanotransduction mode, in which cellular stretch leads to the mechanical formation of membrane templates, which are then reshaped into tubules by BAR proteins. Our results demonstrate the interplay between membrane mechanics and BAR protein molecular organization, integrating curvature sensing and generation in a comprehensive framework with implications for cell mechanical responses.

320 Desmoglein 3-mediated mechanical transadhesion controls epithelial stem cell quiescence

Cell-cell adhesion plays a critical role in the mechanochemical feedback control of tissue homeostasis and disease. However, in mammalian epithelial SC compartments the contribution of classical cadherins in controlling stem cell (SC) quiescence known from lower vertebrates has remained unmatched. We here demonstrate that mechanical uncoupling of desmosomal cadherin desmoglein 3 (Dsg3) by experimental pemphigus vulgaris antibodies activates quiescent SCs in the multipotent hair follicle compartment known as the bulge.

Mechano-chemical feedback mediated competition for BMP signalling leads to pattern formation

Developmental patterning is thought to be regulated by conserved signalling pathways. Initial patterns are often broad before refining to only those cells that commit to a particular fate. However, the mechanisms by which pattern refinement takes place remain to be addressed. Using the posterior crossvein (PCV) of the Drosophila pupal wing as a model, into which bone morphogenetic protein (BMP) ligand is extracellularly transported to instruct vein patterning, we investigate how pattern refinement is regulated. We found that BMP signalling induces apical enrichment of Myosin II in developing crossvein cells to regulate apical constriction. Live imaging of cellular behaviour indicates that changes in cell shape are dynamic and transient, only being maintained in those cells that retain vein fate competence after refinement. Disrupting cell shape changes throughout the PCV inhibits pattern refinement. In contrast, disrupting cell shape in only a subset of vein cells can result in a loss of BMP signalling. We propose that mechano-chemical feedback leads to competition for the developmental signal which plays a critical role in pattern refinement.

Retrograde ERK activation waves drive base-to-apex multicellular flow in murine cochlear duct morphogenesis.

A notable example of spiral architecture in organs is the mammalian cochlear duct, where the morphology is critical for hearing function. Genetic studies have revealed necessary signaling molecules, but it remains unclear how cellular dynamics generate elongating, bending, and coiling of the cochlear duct. Here, we show that extracellular signal-regulated kinase (ERK) activation waves control collective cell migration during the murine cochlear duct development using deep tissue live-cell imaging, Förster resonance energy transfer (FRET)-based quantitation, and mathematical modeling. Long-term FRET imaging reveals that helical ERK activation propagates from the apex duct tip concomitant with the reverse multicellular flow on the lateral side of the developing cochlear duct, resulting in advection-based duct elongation. Moreover, model simulations, together with experiments, explain that the oscillatory wave trains of ERK activity and the cell flow are generated by mechanochemical feedback. Our findings propose a regulatory mechanism to coordinate the multicellular behaviors underlying the duct elongation during development.

Mechanochemical control of epidermal stem cell divisions by B-plexins

The precise spatiotemporal control of cell proliferation is key to the morphogenesis of epithelial tissues. Epithelial cell divisions lead to tissue crowding and local changes in force distribution, which in turn suppress the rate of cell divisions. However, the molecular mechanisms underlying this mechanical feedback are largely unclear. Here, we identify a critical requirement of B-plexin transmembrane receptors in the response to crowding-induced mechanical forces during embryonic skin development. Epidermal stem cells lacking B-plexins fail to sense mechanical compression, resulting in disinhibition of the transcriptional coactivator YAP, hyperproliferation, and tissue overgrowth. Mechanistically, we show that B-plexins mediate mechanoresponses to crowding through stabilization of adhesive cell junctions and lowering of cortical stiffness. Finally, we provide evidence that the B-plexin-dependent mechanochemical feedback is also pathophysiologically relevant to limit tumor growth in basal cell carcinoma, the most common type of skin cancer. Our data define a central role of B-plexins in mechanosensation to couple cell density and cell division in development and disease.

Mechanochemical feedback mediates tissue bending required for seedling emergence

Tissue bending is vital to plant development, as exemplified by apical hook formation during seedling emergence by bending of the hypocotyl. How tissue bending is coordinated during development remains poorly understood, especially in plants where cells are attached via rigid cell walls. Asymmetric distribution of the plant hormone auxin underlies differential cell elongation during apical hook formation. Yet the underlying mechanism remains unclear. Here, we demonstrate spatial correlation between asymmetric auxin distribution, methylesterified homogalacturonan (HG) pectin, and mechanical properties of the epidermal layer ofthe hypocotyl in Arabidopsis. Genetic and cell biological approaches show that this mechanochemical asymmetry is essential for differential cell elongation. We show that asymmetric auxin distribution underlies differential HG methylesterification, and conversely changes in HG methylesterification impact the auxin response domain. Our results suggest that a positive feedback loop between auxin distribution and HG methylesterification underpins asymmetric cell wall mechanochemical properties to promote tissue bending and seedling emergence.

Pattern formation in reaction\u2013diffusion system on membrane with mechanochemical feedback

Shapes of biological membranes are dynamically regulated in living cells. Although membrane shape deformation by proteins at thermal equilibrium has been extensively studied, nonequilibrium dynamics have been much less explored. Recently, chemical reaction propagation has been experimentally observed in plasma membranes. Thus, it is important to understand how the reaction–diffusion dynamics are modified on deformable curved membranes. Here, we investigated nonequilibrium pattern formation on vesicles induced by mechanochemical feedback between membrane deformation and chemical reactions, using dynamically triangulated membrane simulations combined with the Brusselator model. We found that membrane deformation changes stable patterns relative to those that occur on a non-deformable curved surface, as determined by linear stability analysis. We further found that budding and multi-spindle shapes are induced by Turing patterns, and we also observed the transition from oscillation patterns to stable spot patterns. Our results demonstrate the importance of mechanochemical feedback in pattern formation on deforming membranes.

ERK-Mediated Mechanochemical Waves Direct Collective Cell Polarization

During collective migration of epithelial cells, the migration direction is aligned over a tissue-scale expanse. Although the collective cell migration is known to be directed by mechanical forces transmitted via cell-cell junctions, it remains elusive how the intercellular force transmission is coordinated with intracellular biochemical signaling to achieve collective movements. Here, we show that intercellular coupling of extracellular signal-regulated kinase (ERK)-mediated mechanochemical feedback yields long-distance transmission of guidance cues. Mechanical stretch activates ERK through epidermal growth factor receptor (EGFR) activation, and ERK activation triggers cell contraction. The contraction of the activated cell pulls neighboring cells, evoking another round of ERK activation and contraction in the neighbors. Furthermore, anisotropic contraction based on front-rear polarization guarantees unidirectional propagation of ERK activation, and in turn, the ERK activation waves direct multicellular alignment of the polarity, leading to long-range ordered migration. Our findings reveal thatmechanical forces mediate intercellular signaling underlying sustained transmission of guidance cues for collective cell migration.

Cell-substrate mechanics guide collective cell migration through intercellular adhesion: a dynamic finite element cellular model.

During the process of tissue formation and regeneration, cells migrate collectively while remaining connected through intercellular adhesions. However, the roles of cell-substrate and cell-cell mechanical interactions in regulating collective cell migration are still unclear. In this study, we employ a newly developed finite element cellular model to study collective cell migration by exploring the effects of mechanical feedback between cell and substrate and mechanical signal transmission between adjacent cells. Our viscoelastic model of cells consists many triangular elements and is of high resolution. Cadherin adhesion between cells is modeled explicitly as linear springs at subcellular level. In addition, we incorporate a mechano-chemical feedback loop between cell-substrate mechanics and Rac-mediated cell protrusion. Our model can reproduce a number of experimentally observed patterns of collective cell migration during wound healing, including cell migration persistence, separation distance between cell pairs and migration direction. Moreover, we demonstrate that cell protrusion determined by the cell-substrate mechanics plays an important role in guiding persistent and oriented collective cell migration. Furthermore, this guidance cue can be maintained and transmitted to submarginal cells of long distance through intercellular adhesions. Our study illustrates that our finite element cellular model can be employed to study broad problems of complex tissue in dynamic changes at subcellular level.

Fast quantitative time lapse displacement imaging of endothelial cell invasion.

In order to unravel rapid mechano-chemical feedback mechanisms in sprouting angiogenesis, we combine selective plane illumination microscopy (SPIM) and tailored image registration algorithms — further referred to as SPIM-based displacement microscopy — with an in vitro model of angiogenesis. SPIM successfully tackles the problem of imaging large volumes while upholding the spatial resolution required for the analysis of matrix displacements at a subcellular level. Applied to in vitro angiogenic sprouts, this unique methodological combination relates subcellular activity — minute to second time scale growing and retracting of protrusions — of a multicellular systems to the surrounding matrix deformations with an exceptional temporal resolution of 1 minute for a stack with multiple sprouts simultaneously or every 4 seconds for a single sprout, which is 20 times faster than with a conventional confocal setup. Our study reveals collective but non-synchronised, non-continuous activity of adjacent sprouting cells along with correlations between matrix deformations and protrusion dynamics.

Hydrodynamic Synchronization of Spontaneously Beating Filaments.

Using a geometric feedback model of the flagellar axoneme accounting for dynein motor kinetics, we study elastohydrodynamic phase synchronization in a pair of spontaneously beating filaments with waveforms ranging from sperm to cilia and Chlamydomonas. Our computations reveal that both in-phase and antiphase synchrony can emerge for asymmetric beats while symmetric waveforms go in phase, and elucidate the mechanism for phase slips due to biochemical noise. Model predictions agree with recent experiments and illuminate the crucial roles of hydrodynamics and mechanochemical feedback in synchronization.

Biophysics of Cell-Substrate Interactions Under Shear.

Cells adhere to substrates through mechanosensitive focal adhesion complexes. Measurements that probe how cells detach from substrates when they experience an applied force connect molecular-scale aspects of cell adhesion with the biophysical properties of adherent cells. Such forces can be applied through shear devices that flow fluid in a controlled manner across cells. The signaling pathways associated with focal adhesions, in particular those that involve integrins and receptor tyrosine kinases, are complex, receiving mechano-chemical feedback from the sensing of substrate stiffness as well as of external forces. This article reviews the signaling processes involved in mechanosensing and mechanotransduction during cell-substrate interactions, describing the role such signaling plays in cancer metastasis. We examine some recent progress in quantifying the strength of these interactions, describing a novel fluid shear device that allows for the visualization of the cell and its sub-cellular structures under a shear flow. We also summarize related results from a biophysical model for cellular de-adhesion induced by applied forces. Quantifying cell-substrate adhesions under shear should aid in the development of mechano-diagnostic techniques for diseases in which cell-adhesion is mis-regulated, such as cancers.

Mechanochemical self-organization determines search pattern in migratory cells

To reach their destination, migrating cells rely on polarized signal inputs to align the direction of their motion. However, navigational guidance cues are not always present, which establishes the need for exploratory search mechanisms in cells seeking signal inputs. Here, we investigate how non-Brownian search patterns emerge in adherent vertebrate cells. Combining experimental and theoretical analysis, we demonstrate that nanoscale plasma membrane deformations nucleate a mechanochemical feedback loop that mediates longevity of the cell’s leading edge, a necessary requirement for directed cell migration. We further observe stochastic transitions between phases of random and persistent cell motion, whereby the mechanochemical circuit augments cell persistence and search area. Collectively, these findings are consistent with a self-organizing system for a superdiffusive pattern of motion that is spontaneously employed by migratory cells in the absence of external signal inputs. A study of how migrating cells optimize their search efficiency in the absence of directional cues reveals a self-organizing system that mediates superdiffusive motion—and sheds light on how cells navigate noisy environments.

A simple mechanochemical model for calcium signalling in embryonic epithelial cells

Calcium signalling is one of the most important mechanisms of information propagation in the body. In embryogenesis the interplay between calcium signalling and mechanical forces is critical to the healthy development of an embryo but poorly understood. Several types of embryonic cells exhibit calcium-induced contractions and many experiments indicate that calcium signals and contractions are coupled via a two-way mechanochemical feedback mechanism. We present a new analysis of experimental data that supports the existence of this coupling during apical constriction. We then propose a simple mechanochemical model, building on early models that couple calcium dynamics to the cell mechanics and we replace the hypothetical bistable calcium release with modern, experimentally validated calcium dynamics. We assume that the cell is a linear, viscoelastic material and we model the calcium-induced contraction stress with a Hill function, i.e. saturating at high calcium levels. We also express, for the first time, the “stretch-activation” calcium flux in the early mechanochemical models as a bottom-up contribution from stretch-sensitive calcium channels on the cell membrane. We reduce the model to three ordinary differential equations and analyse its bifurcation structure semi-analytically as two bifurcation parameters vary—the textit{IP}_3 concentration, and the “strength” of stretch activation, lambda . The calcium system (lambda =0, no mechanics) exhibits relaxation oscillations for a certain range of textit{IP}_3 values. As lambda is increased the range of textit{IP}_3 values decreases and oscillations eventually vanish at a sufficiently high value of lambda . This result agrees with experimental evidence in embryonic cells which also links the loss of calcium oscillations to embryo abnormalities. Furthermore, as lambda is increased the oscillation amplitude decreases but the frequency increases. Finally, we also identify the parameter range for oscillations as the mechanical responsiveness factor of the cytosol increases. This work addresses a very important and not well studied question regarding the coupling between chemical and mechanical signalling in embryogenesis.

Self-organized shape dynamics of active surfaces

Mechanochemical processes in thin biological structures, such as the cellular cortex or epithelial sheets, play a key role during the morphogenesis of cells and tissues. In particular, they are responsible for the dynamical organization of active stresses that lead to flows and deformations of the material. Consequently, advective transport redistributes force-generating molecules and thereby contributes to a complex mechanochemical feedback loop. It has been shown in fixed geometries that this mechanism enables patterning, but the interplay of these processes with shape changes of the material remains to be explored. In this work, we study the fully self-organized shape dynamics using the theory of active fluids on deforming surfaces and develop a numerical approach to solve the corresponding force and torque balance equations. We describe the spontaneous generation of nontrivial surface shapes, shape oscillations, and directed surface flows that resemble peristaltic waves from self-organized, mechanochemical processes on the deforming surface. Our approach provides opportunities to explore the dynamics of self-organized active surfaces and can help to understand the role of shape as an integral element of the mechanochemical organization of morphogenetic processes.

Mechanochemical feedback control of dynamin independent endocytosis modulates membrane tension in adherent cells

Plasma membrane tension regulates many key cellular processes. It is modulated by, and can modulate, membrane trafficking. However, the cellular pathway(s) involved in this interplay is poorly understood. Here we find that, among a number of endocytic processes operating simultaneously at the cell surface, a dynamin independent pathway, the CLIC/GEEC (CG) pathway, is rapidly and specifically upregulated upon a sudden reduction of tension. Moreover, inhibition (activation) of the CG pathway results in lower (higher) membrane tension. However, alteration in membrane tension does not directly modulate CG endocytosis. This requires vinculin, a mechano-transducer recruited to focal adhesion in adherent cells. Vinculin acts by controlling the levels of a key regulator of the CG pathway, GBF1, at the plasma membrane. Thus, the CG pathway directly regulates membrane tension and is in turn controlled via a mechano-chemical feedback inhibition, potentially leading to homeostatic regulation of membrane tension in adherent cells.

Membrane shape-mediated wave propagation of cortical protein dynamics

Immune cells exhibit stimulation-dependent traveling waves in the cortex, much faster than typical cortical actin waves. These waves reflect rhythmic assembly of both actin machinery and peripheral membrane proteins such as F-BAR domain-containing proteins. Combining theory and experiments, we develop a mechanochemical feedback model involving membrane shape changes and F-BAR proteins that render the cortex an interesting dynamical system. We show that such cortical dynamics manifests itself as ultrafast traveling waves of cortical proteins, in which the curvature sensitivity-driven feedback always constrains protein lateral diffusion in wave propagation. The resulting protein wave propagation mainly reflects the spatial gradient in the timing of local protein recruitment from cytoplasm. We provide evidence that membrane undulations accompany these protein waves and potentiate their propagation. Therefore, membrane shape change and protein curvature sensitivity may have underappreciated roles in setting high-speed cortical signal transduction rhythms.