TY - JOUR AB - In development, lineage segregation is coordinated in time and space. An important example is the mammalian inner cell mass, in which the primitive endoderm (PrE, founder of the yolk sac) physically segregates from the epiblast (EPI, founder of the fetus). While the molecular requirements have been well studied, the physical mechanisms determining spatial segregation between EPI and PrE remain elusive. Here, we investigate the mechanical basis of EPI and PrE sorting. We find that rather than the differences in static cell surface mechanical parameters as in classical sorting models, it is the differences in surface fluctuations that robustly ensure physical lineage sorting. These differential surface fluctuations systematically correlate with differential cellular fluidity, which we propose together constitute a non-equilibrium sorting mechanism for EPI and PrE lineages. By combining experiments and modeling, we identify cell surface dynamics as a key factor orchestrating the correct spatial segregation of the founder embryonic lineages. AU - Yanagida, Ayaka AU - Corujo-Simon, Elena AU - Revell, Christopher K. AU - Sahu, Preeti AU - Stirparo, Giuliano G. AU - Aspalter, Irene M. AU - Winkel, Alex K. AU - Peters, Ruby AU - De Belly, Henry AU - Cassani, Davide A.D. AU - Achouri, Sarra AU - Blumenfeld, Raphael AU - Franze, Kristian AU - Hannezo, Edouard B AU - Paluch, Ewa K. AU - Nichols, Jennifer AU - Chalut, Kevin J. ID - 10825 IS - 5 JF - Cell SN - 00928674 TI - Cell surface fluctuations regulate early embryonic lineage sorting VL - 185 ER - TY - JOUR AB - Embryo development requires biochemical signalling to generate patterns of cell fates and active mechanical forces to drive tissue shape changes. However, how these processes are coordinated, and how tissue patterning is preserved despite the cellular flows occurring during morphogenesis, remains poorly understood. Gastrulation is a crucial embryonic stage that involves both patterning and internalization of the mesendoderm germ layer tissue. Here we show that, in zebrafish embryos, a gradient in Nodal signalling orchestrates pattern-preserving internalization movements by triggering a motility-driven unjamming transition. In addition to its role as a morphogen determining embryo patterning, graded Nodal signalling mechanically subdivides the mesendoderm into a small fraction of highly protrusive leader cells, able to autonomously internalize via local unjamming, and less protrusive followers, which need to be pulled inwards by the leaders. The Nodal gradient further enforces a code of preferential adhesion coupling leaders to their immediate followers, resulting in a collective and ordered mode of internalization that preserves mesendoderm patterning. Integrating this dual mechanical role of Nodal signalling into minimal active particle simulations quantitatively predicts both physiological and experimentally perturbed internalization movements. This provides a quantitative framework for how a morphogen-encoded unjamming transition can bidirectionally couple tissue mechanics with patterning during complex three-dimensional morphogenesis. AU - Nunes Pinheiro, Diana C AU - Kardos, Roland AU - Hannezo, Edouard B AU - Heisenberg, Carl-Philipp J ID - 12209 IS - 12 JF - Nature Physics KW - General Physics and Astronomy SN - 1745-2473 TI - Morphogen gradient orchestrates pattern-preserving tissue morphogenesis via motility-driven unjamming VL - 18 ER - TY - JOUR AB - The development dynamics and self-organization of glandular branched epithelia is of utmost importance for our understanding of diverse processes ranging from normal tissue growth to the growth of cancerous tissues. Using single primary murine pancreatic ductal adenocarcinoma (PDAC) cells embedded in a collagen matrix and adapted media supplementation, we generate organoids that self-organize into highly branched structures displaying a seamless lumen connecting terminal end buds, replicating in vivo PDAC architecture. We identify distinct morphogenesis phases, each characterized by a unique pattern of cell invasion, matrix deformation, protein expression, and respective molecular dependencies. We propose a minimal theoretical model of a branching and proliferating tissue, capturing the dynamics of the first phases. Observing the interaction of morphogenesis, mechanical environment and gene expression in vitro sets a benchmark for the understanding of self-organization processes governing complex organoid structure formation processes and branching morphogenesis. AU - Randriamanantsoa, S. AU - Papargyriou, A. AU - Maurer, H. C. AU - Peschke, K. AU - Schuster, M. AU - Zecchin, G. AU - Steiger, K. AU - Öllinger, R. AU - Saur, D. AU - Scheel, C. AU - Rad, R. AU - Hannezo, Edouard B AU - Reichert, M. AU - Bausch, A. R. ID - 12217 JF - Nature Communications KW - General Physics and Astronomy KW - General Biochemistry KW - Genetics and Molecular Biology KW - General Chemistry KW - Multidisciplinary SN - 2041-1723 TI - Spatiotemporal dynamics of self-organized branching in pancreas-derived organoids VL - 13 ER - TY - JOUR AB - The sculpting of germ layers during gastrulation relies on the coordinated migration of progenitor cells, yet the cues controlling these long-range directed movements remain largely unknown. While directional migration often relies on a chemokine gradient generated from a localized source, we find that zebrafish ventrolateral mesoderm is guided by a self-generated gradient of the initially uniformly expressed and secreted protein Toddler/ELABELA/Apela. We show that the Apelin receptor, which is specifically expressed in mesodermal cells, has a dual role during gastrulation, acting as a scavenger receptor to generate a Toddler gradient, and as a chemokine receptor to sense this guidance cue. Thus, we uncover a single receptor–based self-generated gradient as the enigmatic guidance cue that can robustly steer the directional migration of mesoderm through the complex and continuously changing environment of the gastrulating embryo. AU - Stock, Jessica AU - Kazmar, Tomas AU - Schlumm, Friederike AU - Hannezo, Edouard B AU - Pauli, Andrea ID - 12253 IS - 37 JF - Science Advances SN - 2375-2548 TI - A self-generated Toddler gradient guides mesodermal cell migration VL - 8 ER - TY - JOUR AB - Cell migration in confining physiological environments relies on the concerted dynamics of several cellular components, including protrusions, adhesions with the environment, and the cell nucleus. However, it remains poorly understood how the dynamic interplay of these components and the cell polarity determine the emergent migration behavior at the cellular scale. Here, we combine data-driven inference with a mechanistic bottom-up approach to develop a model for protrusion and polarity dynamics in confined cell migration, revealing how the cellular dynamics adapt to confining geometries. Specifically, we use experimental data of joint protrusion-nucleus migration trajectories of cells on confining micropatterns to systematically determine a mechanistic model linking the stochastic dynamics of cell polarity, protrusions, and nucleus. This model indicates that the cellular dynamics adapt to confining constrictions through a switch in the polarity dynamics from a negative to a positive self-reinforcing feedback loop. Our model further reveals how this feedback loop leads to stereotypical cycles of protrusion-nucleus dynamics that drive the migration of the cell through constrictions. These cycles are disrupted upon perturbation of cytoskeletal components, indicating that the positive feedback is controlled by cellular migration mechanisms. Our data-driven theoretical approach therefore identifies polarity feedback adaptation as a key mechanism in confined cell migration. AU - Brückner, David AU - Schmitt, Matthew AU - Fink, Alexandra AU - Ladurner, Georg AU - Flommersfeld, Johannes AU - Arlt, Nicolas AU - Hannezo, Edouard B AU - Rädler, Joachim O. AU - Broedersz, Chase P. ID - 12277 IS - 3 JF - Physical Review X KW - General Physics and Astronomy SN - 2160-3308 TI - Geometry adaptation of protrusion and polarity dynamics in confined cell migration VL - 12 ER - TY - JOUR AB - The morphology and functionality of the epithelial lining differ along the intestinal tract, but tissue renewal at all sites is driven by stem cells at the base of crypts1,2,3. Whether stem cell numbers and behaviour vary at different sites is unknown. Here we show using intravital microscopy that, despite similarities in the number and distribution of proliferative cells with an Lgr5 signature in mice, small intestinal crypts contain twice as many effective stem cells as large intestinal crypts. We find that, although passively displaced by a conveyor-belt-like upward movement, small intestinal cells positioned away from the crypt base can function as long-term effective stem cells owing to Wnt-dependent retrograde cellular movement. By contrast, the near absence of retrograde movement in the large intestine restricts cell repositioning, leading to a reduction in effective stem cell number. Moreover, after suppression of the retrograde movement in the small intestine, the number of effective stem cells is reduced, and the rate of monoclonal conversion of crypts is accelerated. Together, these results show that the number of effective stem cells is determined by active retrograde movement, revealing a new channel of stem cell regulation that can be experimentally and pharmacologically manipulated. AU - Azkanaz, Maria AU - Corominas-Murtra, Bernat AU - Ellenbroek, Saskia I. J. AU - Bruens, Lotte AU - Webb, Anna T. AU - Laskaris, Dimitrios AU - Oost, Koen C. AU - Lafirenze, Simona J. A. AU - Annusver, Karl AU - Messal, Hendrik A. AU - Iqbal, Sharif AU - Flanagan, Dustin J. AU - Huels, David J. AU - Rojas-Rodríguez, Felipe AU - Vizoso, Miguel AU - Kasper, Maria AU - Sansom, Owen J. AU - Snippert, Hugo J. AU - Liberali, Prisca AU - Simons, Benjamin D. AU - Katajisto, Pekka AU - Hannezo, Edouard B AU - van Rheenen, Jacco ID - 12274 IS - 7919 JF - Nature KW - Multidisciplinary SN - 0028-0836 TI - Retrograde movements determine effective stem cell numbers in the intestine VL - 607 ER - TY - GEN AB - Source data and source code for the graphs in "Spatiotemporal dynamics of self-organized branching pancreatic cancer-derived organoids". AU - Randriamanantsoa, Samuel AU - Papargyriou, Aristeidis AU - Maurer, Carlo AU - Peschke, Katja AU - Schuster, Maximilian AU - Zecchin, Giulia AU - Steiger, Katja AU - Öllinger, Rupert AU - Saur, Dieter AU - Scheel, Christina AU - Rad, Roland AU - Hannezo, Edouard B AU - Reichert, Maximilian AU - Bausch, Andreas R. ID - 13068 TI - Spatiotemporal dynamics of self-organized branching in pancreas-derived organoids ER - TY - JOUR AB - Collective cell migration offers a rich field of study for non-equilibrium physics and cellular biology, revealing phenomena such as glassy dynamics, pattern formation and active turbulence. However, how mechanical and chemical signalling are integrated at the cellular level to give rise to such collective behaviours remains unclear. We address this by focusing on the highly conserved phenomenon of spatiotemporal waves of density and extracellular signal-regulated kinase (ERK) activation, which appear both in vitro and in vivo during collective cell migration and wound healing. First, we propose a biophysical theory, backed by mechanical and optogenetic perturbation experiments, showing that patterns can be quantitatively explained by a mechanochemical coupling between active cellular tensions and the mechanosensitive ERK pathway. Next, we demonstrate how this biophysical mechanism can robustly induce long-ranged order and migration in a desired orientation, and we determine the theoretically optimal wavelength and period for inducing maximal migration towards free edges, which fits well with experimentally observed dynamics. We thereby provide a bridge between the biophysical origin of spatiotemporal instabilities and the design principles of robust and efficient long-ranged migration. AU - Boocock, Daniel R AU - Hino, Naoya AU - Ruzickova, Natalia AU - Hirashima, Tsuyoshi AU - Hannezo, Edouard B ID - 8602 JF - Nature Physics SN - 17452473 TI - Theory of mechanochemical patterning and optimal migration in cell monolayers VL - 17 ER - TY - JOUR AB - Organ function depends on tissues adopting the correct architecture. However, insights into organ architecture are currently hampered by an absence of standardized quantitative 3D analysis. We aimed to develop a robust technology to visualize, digitalize, and segment the architecture of two tubular systems in 3D: double resin casting micro computed tomography (DUCT). As proof of principle, we applied DUCT to a mouse model for Alagille syndrome (Jag1Ndr/Ndr mice), characterized by intrahepatic bile duct paucity, that can spontaneously generate a biliary system in adulthood. DUCT identified increased central biliary branching and peripheral bile duct tortuosity as two compensatory processes occurring in distinct regions of Jag1Ndr/Ndr liver, leading to full reconstitution of wild-type biliary volume and phenotypic recovery. DUCT is thus a powerful new technology for 3D analysis, which can reveal novel phenotypes and provide a standardized method of defining liver architecture in mouse models. AU - Hankeova, Simona AU - Salplachta, Jakub AU - Zikmund, Tomas AU - Kavkova, Michaela AU - Van Hul, Noémi AU - Brinek, Adam AU - Smekalova, Veronika AU - Laznovsky, Jakub AU - Dawit, Feven AU - Jaros, Josef AU - Bryja, Vítězslav AU - Lendahl, Urban AU - Ellis, Ewa AU - Nemeth, Antal AU - Fischler, Björn AU - Hannezo, Edouard B AU - Kaiser, Jozef AU - Andersson, Emma Rachel ID - 9244 JF - eLife TI - DUCT reveals architectural mechanisms contributing to bile duct recovery in a mouse model for alagille syndrome VL - 10 ER - TY - JOUR AB - Assemblies of actin and its regulators underlie the dynamic morphology of all eukaryotic cells. To understand how actin regulatory proteins work together to generate actin-rich structures such as filopodia, we analyzed the localization of diverse actin regulators within filopodia in Drosophila embryos and in a complementary in vitro system of filopodia-like structures (FLSs). We found that the composition of the regulatory protein complex where actin is incorporated (the filopodial tip complex) is remarkably heterogeneous both in vivo and in vitro. Our data reveal that different pairs of proteins correlate with each other and with actin bundle length, suggesting the presence of functional subcomplexes. This is consistent with a theoretical framework where three or more redundant subcomplexes join the tip complex stochastically, with any two being sufficient to drive filopodia formation. We provide an explanation for the observed heterogeneity and suggest that a mechanism based on multiple components allows stereotypical filopodial dynamics to arise from diverse upstream signaling pathways. AU - Dobramysl, Ulrich AU - Jarsch, Iris Katharina AU - Inoue, Yoshiko AU - Shimo, Hanae AU - Richier, Benjamin AU - Gadsby, Jonathan R. AU - Mason, Julia AU - Szałapak, Alicja AU - Ioannou, Pantelis Savvas AU - Correia, Guilherme Pereira AU - Walrant, Astrid AU - Butler, Richard AU - Hannezo, Edouard B AU - Simons, Benjamin D. AU - Gallop, Jennifer L. ID - 9306 IS - 4 JF - Journal of Cell Biology TI - Stochastic combinations of actin regulatory proteins are sufficient to drive filopodia formation VL - 220 ER -