Multicellular Patterns Research Articles

Coordinated, multicellular patterns of transcriptional variation that stratify patient cohorts are revealed by tensor decomposition.

Tissue-level and organism-level biological processes often involve the coordinated action of multiple distinct cell types. The recent application of single-cell assays to many individuals should enable the study of how donor-level variation in one cell type is linked to that in other cell types. Here we introduce a computational approach called single-cell interpretable tensor decomposition (scITD) to identify common axes of interindividual variation by considering joint expression variation across multiple cell types. scITD combines expression matrices from each cell type into a higher-order matrix and factorizes the result using the Tucker tensor decomposition. Applying scITD to single-cell RNA-sequencing data on 115 persons with lupus and 83 persons with coronavirus disease 2019, we identify patterns of coordinated cellular activity linked to disease severity and specific phenotypes, such as lupus nephritis. scITD results also implicate specific signaling pathways likely mediating coordination between cell types. Overall, scITD offers a tool for understanding the covariation of cell states across individuals, which can yield insights into the complex processes that define and stratify disease.

Rapid and Controllable Multilayer Cell Sheet Assembly via Biodegradable Nanochannel Membranes

AbstractThe ability to precisely arrange and control the assembly of diverse cell types into intricate 3D structures remains a critical challenge in tissue engineering. Herein, a versatile and programmable 3D cell sheet assembly is described technology by developing a biodegradable nanochannel (BNC) membrane to fulfill this unmet need. This membrane, hierarchically assembled from 2D nanomaterial aggregates, exhibits both exceptional fluid permeability and rapid biodegradation under physiological conditions. The unique properties of the BNC membrane enable precise spatial and temporal control over cell assembly, facilitating the creation of complex 3D cellular architectures. The BNC membrane is integrated with a programmable negative‐pressure‐based cell assembly strategy to form single and multicellular 3D sheets in a highly controllable manner. To demonstrate the feasibility and translatability of this technology in the field of tissue engineering approaches to screen stem cell‐derived therapeutics with “core–shell” macrophage‐fibroblast multicellular patterns and treat murine diabetic skin wounds via scaffold‐free 3D adipose‐derived mesenchymal stem cell (ADMSC) sheets are devised. In summary, the results demonstrate that the BNC membrane‐based 3D cell sheet assembly approach significantly advances current tissue engineering capabilities, offering substantial potential for both regenerative medicine applications and the development of physiologically relevant disease models.

A mathematical framework for understanding the spontaneous emergence of complexity applicable to growing multicellular systems.

In embryonic development and organogenesis, cells sharing identical genetic codes acquire diverse gene expression states in a highly reproducible spatial distribution, crucial for multicellular formation and quantifiable through positional information. To understand the spontaneous growth of complexity, we constructed a one-dimensional division-decision model, simulating the growth of cells with identical genetic networks from a single cell. Our findings highlight the pivotal role of cell division in providing positional cues, escorting the system toward states rich in information. Moreover, we pinpointed lateral inhibition as a critical mechanism translating spatial contacts into gene expression. Our model demonstrates that the spatial arrangement resulting from cell division, combined with cell lineages, imparts positional information, specifying multiple cell states with increased complexity-illustrated through examples in C.elegans. This study constitutes a foundational step in comprehending developmental intricacies, paving the way for future quantitative formulations to construct synthetic multicellular patterns.

Agent-based modeling of stress anisotropy driven nematic ordering in growing biofilms.

Living active collectives have evolved with remarkable self-patterning capabilities to adapt to the physical and biological constraints crucial for their growth and survival. However, the intricate process by which complex multicellular patterns emerge from a single founder cell remains elusive. In this study, we utilize an agent-based model, validated through single-cell microscopy imaging, to track the three-dimensional (3D) morphodynamics of cells within growing bacterial biofilms encased by agarose gels. The confined growth conditions give rise to a spatiotemporally heterogeneous stress landscape within the biofilm. In the core of the biofilm, where high hydrostatic and low shear stresses prevail, cell packing appears disordered. In contrast, near the gel-cell interface, a state of high shear stress and low hydrostatic stress emerges, driving nematic ordering, albeit with a time delay inherent to shear stress relaxation. Strikingly, we observe a robust spatiotemporal correlation between stress anisotropy and nematic ordering within these confined biofilms. This correlation suggests a mechanism whereby stress anisotropy plays a pivotal role in governing the spatial organization of cells. The reciprocity between stress anisotropy and cell ordering in confined biofilms opens new avenues for innovative 3D mechanically guided patterning techniques for living active collectives, which hold significant promise for a wide array of environmental and biomedical applications.

Strength in numbers: Unleashing the potential of trans-scale scope AMATERAS for massive cell quantification.

Singularity biology is a scientific field that targets drastic state changes in multicellular systems, aiming to discover the key cells that induce the state change and investigate the mechanisms behind them. To achieve this goal, we developed a trans-scale optical imaging system (trans-scale scope), that is capable of capturing both macroscale changes across the entire system and the micro-scale behavior of individual cells, surpassing the cell observation capabilities of traditional microscopes. We developed two units of the trans-scale scope, named AMATERAS-1 and -2, which demonstrated the ability to observe multicellular systems consisting of over one million cells in a single field of view with sub-cellular resolution. This flagship instrument has been used to observe the dynamics of various cell species, with the advantage of being able to observe a large number of cells, allowing the detection and analysis of rare events and cells such as leader cells in multicellular pattern formation and cells that spontaneously initiate calcium waves. In this paper, we present the design concept of AMATERAS, the optical configuration, and several examples of observations, and demonstrate how the strength-in-numbers works in life sciences.

Topological data analysis of spatial patterning in heterogeneous cell populations: clustering and sorting with varying cell-cell adhesion

Different cell types aggregate and sort into hierarchical architectures during the formation of animal tissues. The resulting spatial organization depends (in part) on the strength of adhesion of one cell type to itself relative to other cell types. However, automated and unsupervised classification of these multicellular spatial patterns remains challenging, particularly given their structural diversity and biological variability. Recent developments based on topological data analysis are intriguing to reveal similarities in tissue architecture, but these methods remain computationally expensive. In this article, we show that multicellular patterns organized from two interacting cell types can be efficiently represented through persistence images. Our optimized combination of dimensionality reduction via autoencoders, combined with hierarchical clustering, achieved high classification accuracy for simulations with constant cell numbers. We further demonstrate that persistence images can be normalized to improve classification for simulations with varying cell numbers due to proliferation. Finally, we systematically consider the importance of incorporating different topological features as well as information about each cell type to improve classification accuracy. We envision that topological machine learning based on persistence images will enable versatile and robust classification of complex tissue architectures that occur in development and disease.

Characterizing and tuning the bacterial synthetic adhesion biophysics for biomaterial applications.

Bacterial synthetic multi-cellular (SMC) systems present promising characteristics as engineering platforms for developing complex products in the areas of medicine, biosynthesis and smart materials. This work has been recently advanced by the development of a genetically encoded toolkit of surface-bound nanobodies and antigens via the intimin autotransporter in E. coli that can enable precise manipulation of cell-cell adhesion and design of self-assembled multicellular patterns. However, it is necessary to be able to predict how adhering cells behave inside synthetic consortia and more specifically, how the adhesins behave on the surface of each of these cells, in order to advance the functionality of the adhesin toolkit at the subcellular level. We quantitatively measured the key biophysical (kinetic) parameters that determine dynamics (and mechanics) of the synthetic adhesins on the cell surface, such as the lateral diffusion coefficient, the production and degradation rates and the binding force between two complementary adhesins. The complete characterization of the adhesin construct enables us to further their application in the field of engineered living materials (ELMs). We show that tuning the adhesin biophysical parameters at a microscopic level can change the biomaterial properties at a macroscopic level. These results are critical for rational engineering and modeling synthetic adhesins and ultimately multicellular synthetic consortia and will also be valuable for researchers working with this intimin and potentially other autotransporter and nanobodies in various other contexts by providing the methodology for the characterization of future synthetic adhesins.

Unorthodox regulation of the MglA Ras-like GTPase controlling polarity in Myxococcus xanthus.

Motile cells have developed a large array of molecular machineries to actively change their direction of movement in response to spatial cues from their environment. In this process, small GTPases act as molecular switches and work in tandem with regulators and sensors of their guanine nucleotide status (GAP, GEF, GDI and effectors) to dynamically polarize the cell and regulate its motility. In this review, we focus on Myxococcus xanthus as a model organism to elucidate the function of an atypical small Ras GTPase system in the control of directed cell motility. M.xanthus cells direct their motility by reversing their direction of movement through a mechanism involving the redirection of the motility apparatus to the opposite cell pole. The reversal frequency of moving M.xanthus cells is controlled by modular and interconnected protein networks linking the chemosensory-like frizzy (Frz) pathway - that transmits environmental signals - to the downstream Ras-like Mgl polarity control system - that comprises the Ras-like MglA GTPase protein and its regulators. Here, we discuss how variations in the GTPase interactome landscape underlie single-cell decisions and consequently, multicellular patterns.

Buoyancy-Induced Instability of a Power-Law Fluid Saturating a Vertical Porous Slab

Abstract Many engineering applications involve porous media and rely on non-Newtonian working fluids. In this paper, the seepage flow of a non-Newtonian fluid saturating a vertical porous layer is studied. The buoyant flow is thermally driven by the boundaries of the porous layer, which are permeable surfaces kept at different temperatures. In order to model the seepage flow of both shear-thinning (pseudoplastic) and shear-thickening (dilatant) fluids, reference is made to the Ostwald-de Waele rheological model implemented via the power-law extended form of Darcy's law. The basic stationary flow is parallel to the vertical axis and shows a single-cell pattern, where the cell has infinite height and can display a core-region of enhanced/inhibited flow according to the fluid's rheological behavior. By applying small perturbations, a linear stability analysis of the basic flow is performed to determine the onset conditions for a multicellular pattern. This analysis is carried out numerically by employing the shooting method. The neutral stability curves and the values of the critical Rayleigh number are computed for different pseudoplastic and dilatant fluids. The behavior of a Newtonian fluid is also obtained as a limiting case.

Cells-Micropatterning Biomaterials for Immune Activation and Bone Regeneration.

Natural tissues are composed of ordered architectural organizations of multiple tissue cells. The spatial distribution of cells is crucial for directing cellular behavior and maintaining tissue homeostasis and function. Herein, an artificial bone bioceramic scaffold with star‐, Tai Chi‐, or interlacing‐shaped multicellular patterns is constructed. The “cross‐talk” between mesenchymal stem cells (MSCs) and macrophages can be effectively manipulated by altering the spatial distribution of two kinds of cells in the scaffolds, thus achieving controllable modulation of the scaffold‐mediated osteo‐immune responses. Compared with other multicellular patterns, the Tai Chi pattern with a 2:1 ratio of MSCs to macrophages is more effective in activating anti‐inflammatory M2 macrophages, improving MSCs osteogenic differentiation, and accelerating new bone formation in vivo. In brief, the Tai Chi pattern generates a more favorable osteo‐immune environment for bone regeneration, exhibiting enhanced immunomodulation and osteogenesis, which may be associated with the activation of BMP‐Smad, Oncostatin M (OSM), and Wnt/β‐catenin signaling pathways in MSCs mediated by macrophage‐derived paracrine signaling mediators. The study suggests that the manipulation of cell distribution to improve tissue formation is a feasible approach that can offer new insights for the design of tissue‐engineered bone substitutes with multicellular interactions.

Stability of a Buoyant Oldroyd-B Flow Saturating a Vertical Porous Layer with Open Boundaries

The performance of several engineering applications are strictly connected to the rheology of the working fluids and the Oldroyd-B model is widely employed to describe a linear viscoelastic behaviour. In the present paper, a buoyant Oldroyd-B flow in a vertical porous layer with permeable and isothermal boundaries is investigated. Seepage flow is modelled through an extended version of Darcy’s law which accounts for the Oldroyd-B rheology. The basic stationary flow is parallel to the vertical axis and describes a single-cell pattern where the cell has an infinite height. A linear stability analysis of such a basic flow is carried out to determine the onset conditions for a multicellular pattern. This analysis is performed numerically by employing the shooting method. The neutral stability curves and the values of the critical Rayleigh number are evaluated for different retardation time and relaxation time characteristics of the fluid. The study highlights the extent to which the viscoelasticity has a destabilising effect on the buoyant flow. For the limiting case of a Newtonian fluid, the known results available in the literature are recovered, namely a critical value of the Darcy–Rayleigh number equal to 197.081 and a corresponding critical wavenumber of 1.05950.

Profiling Myxococcus xanthus Swarming Phenotypes through Mutation and Environmental Variation.

ABSTRACTMyxococcus xanthus is a bacterium that lives on surfaces as a predatory biofilm called a swarm. As a growing swarm feeds on prey and expands, it displays dynamic multicellular patterns such as traveling waves called ripples and branching protrusions called flares. The rate at which a swarm expands across a surface, and the emergence of the coexisting patterns, are all controlled through coordinated cell movement. M. xanthus cells move using two motility systems known as adventurous (A) and social (S). Both are involved in swarm expansion and pattern formation. In this study, we describe a set of M. xanthus swarming genotype-to-phenotype associations that include both genetic and environmental perturbations. We identified new features of the swarming phenotype, recorded and measured swarm expansion using time-lapse microscopy, and compared the impact of mutations on different surfaces. These observations and analyses have increased our ability to discriminate between swarming phenotypes and provided context that allows us to identify some phenotypes as improbable outliers within the M. xanthus swarming phenome.IMPORTANCE Myxococcus xanthus grows on surfaces as a predatory biofilm called a swarm. In nature, a feeding swarm expands by moving over and consuming prey bacteria. In the laboratory, a swarm is created by spotting cell suspension onto nutrient agar in lieu of prey. The suspended cells quickly settle on the surface as the liquid is absorbed into the agar, and the new swarm then expands radially. An assay that measures the expansion rate of a swarm of mutant cells is the first, and sometimes only, measurement used to decide whether a particular mutation impacts swarm motility. We have broadened the scope of this assay by increasing the accuracy of measurements and introducing prey, resulting in new identifiable and quantifiable features that can be used to improve genotype-to-phenotype associations.

Weakening of resistance force by cell\u2013ECM interactions regulate cell migration directionality and pattern formation

Collective migration of epithelial cells is a fundamental process in multicellular pattern formation. As they expand their territory, cells are exposed to various physical forces generated by cell–cell interactions and the surrounding microenvironment. While the physical stress applied by neighbouring cells has been well studied, little is known about how the niches that surround cells are spatio-temporally remodelled to regulate collective cell migration and pattern formation. Here, we analysed how the spatio-temporally remodelled extracellular matrix (ECM) alters the resistance force exerted on cells so that the cells can expand their territory. Multiple microfabrication techniques, optical tweezers, as well as mathematical models were employed to prove the simultaneous construction and breakage of ECM during cellular movement, and to show that this modification of the surrounding environment can guide cellular movement. Furthermore, by artificially remodelling the microenvironment, we showed that the directionality of collective cell migration, as well as the three-dimensional branch pattern formation of lung epithelial cells, can be controlled. Our results thus confirm that active remodelling of cellular microenvironment modulates the physical forces exerted on cells by the ECM, which contributes to the directionality of collective cell migration and consequently, pattern formation.

Combinatorial patterns of graded RhoA activation and uniform F-actin depletion promote tissue curvature.

During development, gene expression regulates cell mechanics and shape to sculpt tissues. Epithelial folding proceeds through distinct cell shape changes that occur simultaneously in different regions of a tissue. Here, using quantitative imaging in Drosophila melanogaster, we investigate how patterned cell shape changes promote tissue bending during early embryogenesis. We find that the transcription factors Twist and Snail combinatorially regulate a multicellular pattern of lateral F-actin density that differs from the previously described Myosin-2 gradient. This F-actin pattern correlates with whether cells apically constrict, stretch or maintain their shape. We show that the Myosin-2 gradient and F-actin depletion do not depend on force transmission, suggesting that transcriptional activity is required to create these patterns. The Myosin-2 gradient width results from a gradient in RhoA activation that is refined through the balance between RhoGEF2 and the RhoGAP C-GAP. Our experimental results and simulations of a 3D elastic shell model show that tuning gradient width regulates tissue curvature.

Physical Interaction of Cells with ECM Weakens the Resistance Force and Regulates the Directionality of Collective Cell Migration and Pattern Formation

Collective migration of epithelial cells is a fundamental process in multi-cellular pattern formation. As they expand their territory, cells are exposed to various physical forces generated by cell-cell interactions and the surrounding microenvironment. While the physical stress applied by neighbouring cells has been well studied, little is known about how the niches that surround cells are spatio-temporally remodelled to regulate collective cell migration and pattern formation. Here, we analysed how the spatio-temporally remodelled extracellular matrix (ECM) alters the resistance force exerted on cells so that the cells can expand their territory. Multiple microfabrication techniques, optical tweezers, as well as mathematical models were employed to prove the simultaneous construction and breakage of ECM during cellular movement, and to show that this modification of the surrounding environment can guide cellular movement. Furthermore, by artificially remodelling the microenvironment, we showed that the directionality of collective cell migration, as well as the three dimensional branch pattern formation of lung epithelial cells, can be controlled. Our results thus confirm that active remodelling of cellular microenvironment modulates the physical forces exerted on cells by the ECM, which contributes to the directionality of collective cell migration and consequently, pattern formation.

Dynamic Uni- and Multicellular Patterns Encode Biphasic Activity in Pancreatic Islets.

Biphasic secretion is an autonomous feature of many endocrine micro-organs to fulfill physiological demands. The biphasic activity of islet β-cells maintains glucose homeostasis and is altered in type 2 diabetes. Nevertheless, underlying cellular or multicellular functional organizations are only partially understood. High-resolution noninvasive multielectrode array recordings permit simultaneous analysis of recruitment, of single-cell, and of coupling activity within entire islets in long-time experiments. Using this unbiased approach, we addressed the organizational modes of both first and second phase in mouse and human islets under physiological and pathophysiological conditions. Our data provide a new uni- and multicellular model of islet β-cell activation: during the first phase, small but highly active β-cell clusters are dominant, whereas during the second phase, electrical coupling generates large functional clusters via multicellular slow potentials to favor an economic sustained activity. Postprandial levels of glucagon-like peptide 1 favor coupling only in the second phase, whereas aging and glucotoxicity alter coupled activity in both phases. In summary, biphasic activity is encoded upstream of vesicle pools at the micro-organ level by multicellular electrical signals and their dynamic synchronization between β-cells. The profound alteration of the electrical organization of islets in pathophysiological conditions may contribute to functional deficits in type 2 diabetes.

Data-Driven Models Reveal Mutant Cell Behaviors Important for Myxobacterial Aggregation.

Single mutations frequently alter several aspects of cell behavior but rarely reveal whether a particular statistically significant change is biologically significant. To determine which behavioral changes are most important for multicellular self-organization, we devised a new methodology using Myxococcus xanthus as a model system. During development, myxobacteria coordinate their movement to aggregate into spore-filled fruiting bodies. We investigate how aggregation is restored in two mutants, csgA and pilC, that cannot aggregate unless mixed with wild-type (WT) cells. To this end, we use cell tracking to follow the movement of fluorescently labeled cells in combination with data-driven agent-based modeling. The results indicate that just like WT cells, both mutants bias their movement toward aggregates and reduce motility inside aggregates. However, several aspects of mutant behavior remain uncorrected by WT, demonstrating that perfect recreation of WT behavior is unnecessary. In fact, synergies between errant behaviors can make aggregation robust.IMPORTANCE Self-organization into spatial patterns is evident in many multicellular phenomena. Even for the best-studied systems, our ability to dissect the mechanisms driving coordinated cell movement is limited. While genetic approaches can identify mutations perturbing multicellular patterns, the diverse nature of the signaling cues coupled to significant heterogeneity of individual cell behavior impedes our ability to mechanistically connect genes with phenotype. Small differences in the behaviors of mutant strains could be irrelevant or could sometimes lead to large differences in the emergent patterns. Here, we investigate rescue of multicellular aggregation in two mutant strains of Myxococcus xanthus mixed with wild-type cells. The results demonstrate how careful quantification of cell behavior coupled to data-driven modeling can identify specific motility features responsible for cell aggregation and thereby reveal important synergies and compensatory mechanisms. Notably, mutant cells do not need to precisely recreate wild-type behaviors to achieve complete aggregation.

Radial-velocity variations due to meridional flows in the Sun and solar-type stars: impact on exoplanet detectability

Context. Stellar variability due to magnetic activity and flows at different spatial scales strongly impacts radial velocities. This variability is seen as oscillations, granulation, supergranulation, and meridional flows. The effect of this latter process is currently poorly known but could affect exoplanet detectability. Aims. We aim to quantify the amplitude of the meridional flow integrated over the disc and its temporal variability, first for the Sun, as seen with different inclinations, and then for other solar-type stars. We then want to compare these amplitudes with low-mass exoplanetary amplitudes in radial velocity. Methods. We used long time series (covering two 11-yr cycles) of solar latitudinal meridional circulation to reconstruct its integrated contribution and study its properties. We then used scaling laws from hydrodynamical simulations relating the amplitude of the meridional flow variability with stellar mass and rotation rate to estimate the typical amplitude expected for other solar-type stars. Results. We find typical rms of the order of 0.5–0.7 m s−1 (edge-on) and 1.2–1.7 m s−1 (pole-on) for the Sun (peak-to-peak amplitudes are typically 1–1.4 m s−1 and 2.3–3.3 m s−1 resp.), with a minimal jitter for an inclination of 45–55°. This signal is significant compared to other stellar activity contributions and is much larger than the radial-velocity signal of the Earth. The variability is strongly related to the activity cycle, with maximum flows during the descending phase of the cycle, and possible variability on timescales lower than the cycle period. Extension to other solar-type stars shows that the variability due to meridional flows is dominated by the amplitude of the cycle of those stars (compared with mass and rotation rate), and that the peak-to-peak amplitudes can reach 4 m s−1 for the most variable stars when seen pole-on. The meridional flow contribution sometimes represents a high fraction of the convective blueshift inhibition signal, especially for quiet, low-mass stars. For fast-rotating stars, the presence of multi-cellular patterns should significantly decrease the meridional flow contribution to the radial-velocity signal. Conclusions. Our study shows that these meridional flows could be critical for exoplanet detection. Low inclinations are more impacted than edge-on configurations, but these latter still exhibit significant variability. Meridional flows also degrade the correlation between radial velocities due to convective blueshift inhibition and chromospheric activity indicators. This will make the correction from this signal challenging for stars with no multi-cellular patterns, such as the Sun for example, although there may be some configurations for which the line shape variations may be used if the precision is sufficient.

Bioelectrical Coupling of Single-Cell States in Multicellular Systems.

The spatiotemporal distributions of signaling ions and molecules that modulate biochemical pathways in nonexcitable cells are influenced by multicellular electric potentials. These potentials act as distributed controllers encoding instructive spatial patterns in development and regeneration. We review experimental facts and discuss recent bioelectrical models that provide new physical insights and complement biochemical approaches. Single-cell states are modulated at the multicellular level because of the coupling between neighboring cells, thus allowing memories and multicellular patterns. The model is based on (i) two generic voltage-gated ion channels that promote the polarized and depolarized cell states, (ii) a feedback mechanism for the transcriptional and bioelectrical regulations, and (iii) voltage-gated intercellular conductances that allow a dynamic intercellular connectivity. The simulations provide steady-state and oscillatory multicellular states that help explain aspects of development and guide experimental procedures attempting to establish instructive bioelectrical patterns based on electric potentials and currents to regulate cell behavior and morphogenesis.

Continuous Dynamic Modeling of Regulated Cell Adhesion: Sorting, Intercalation, and Involution

Cell-cell adhesion is essential for tissue growth and multicellular pattern formation and crucial for the cellular dynamics during embryogenesis and cancer progression. Understanding the dynamical gene regulation of cell adhesion molecules (CAMs) responsible for the emerging spatial tissue behaviors is a current challenge because of the complexity of these nonlinear interactions and feedback loops at different levels of abstraction—from genetic regulation to whole-organism shape formation. To extend our understanding of cell and tissue behaviors due to the regulation of adhesion molecules, here we present a novel, to our knowledge, model for the spatial dynamics of cellular patterning, growth, and shape formation due to the differential expression of CAMs and their regulation. Capturing the dynamic interplay between genetic regulation, CAM expression, and differential cell adhesion, the proposed continuous model can explain the complex and emergent spatial behaviors of cell populations that change their adhesion properties dynamically because of inter- and intracellular genetic regulation. This approach can demonstrate the mechanisms responsible for classical cell-sorting behaviors, cell intercalation in proliferating populations, and the involution of germ layer cells induced by a diffusing morphogen during gastrulation. The model makes predictions on the physical parameters controlling the amplitude and wavelength of a cellular intercalation interface, as well as the crucial role of N-cadherin regulation for the involution and migration of cells beyond the gradient of the morphogen Nodal during zebrafish gastrulation. Integrating the emergent spatial tissue behaviors with the regulation of genes responsible for essential cellular properties such as adhesion will pave the way toward understanding the genetic regulation of large-scale complex patterns and shapes formation in developmental, regenerative, and cancer biology.