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Synthetic Notch (synNotch) receptors are genetically encoded, modular synthetic receptors that enable mammalian cells to detect environmental signals and respond by activating user-prescribed transcriptional programs. Although some materials have been modified to present synNotch ligands with coarse spatial control, applications in tissue engineering generally require extracellular matrix (ECM)-derived scaffolds and/or finer spatial positioning of multiple ligands. Thus, we develop here a suite of materials that activate synNotch receptors for generalizable engineering of material-to-cell signaling. We genetically and chemically fuse functional synNotch ligands to ECM proteins and ECM-derived materials. We also generate tissues with microscale precision over four distinct reporter phenotypes by culturing cells with two orthogonal synNotch programs on surfaces microcontact-printed with two synNotch ligands. Finally, we showcase applications in tissue engineering by co-transdifferentiating fibroblasts into skeletal muscle or endothelial cell precursors in user-defined micropatterns. These technologies provide avenues for spatially controlling cellular phenotypes in mammalian tissues. 

We investigate how matrix stiffness regulates chromatin reorganization and cell reprogramming and find that matrix stiffness acts as a biphasic regulator of epigenetic state and fibroblast-to-neuron conversion efficiency, maximized at an intermediate stiffness of 20 kPa. ATAC sequencing analysis shows the same trend of chromatin accessibility to neuronal genes at these stiffness levels. Concurrently, we observe peak levels of histone acetylation and histone acetyltransferase (HAT) activity in the nucleus on 20 kPa matrices, and inhibiting HAT activity abolishes matrix stiffness effects. G-actin and cofilin, the cotransporters shuttling HAT into the nucleus, rises with decreasing matrix stiffness; however, reduced importin-9 on soft matrices limits nuclear transport. These two factors result in a biphasic regulation of HAT transport into nucleus, which is directly demonstrated on matrices with dynamically tunable stiffness. Our findings unravel a mechanism of the mechano-epigenetic regulation that is valuable for cell engineering in disease modeling and regenerative medicine applications. 

Abstract Consumers must track and acquire resources in complex landscapes. Much discussion has focused on the concept of a ‘resource gradient’ and the mechanisms by which consumers can take advantage of such gradients as they navigate their landscapes in search of resources. However, the concept of tracking resource gradients means different things in different contexts. Here, we take a synthetic approach and consider six different definitions of what it means to search for resources based on density or gradients in density. These include scenarios where consumers change their movement behavior based on the density of conspecifics, on the density of resources, and on spatial or temporal gradients in resources. We also consider scenarios involving non-local perception and a form of memory. Using a continuous space, continuous time model that allows consumers to switch between resource-tracking and random motion, we investigate the relative performance of these six different strategies. Consumers’ success in matching the spatiotemporal distributions of their resources differs starkly across the six scenarios. Movement strategies based on perception and response to temporal (rather than spatial) resource gradients afforded consumers with the best opportunities to match resource distributions. All scenarios would allow for optimization of resource-matching in terms of the underlying parameters, providing opportunities for evolutionary adaptation, and links back to classical studies of foraging ecology. 

Perception is central to the survival of an individual for many reasons, especially as it affects the ability to gather resources. Consequently, costs associated with perception are partially shaped by resource availability. Understanding the interplay of environmental factors (such as the density and distribution of resources) with species-specific factors (such as growth rate, mutation, and metabolic costs) allows the exploration of possible trajectories by which perception may evolve. Here, we used an agent-based foraging model with a context-dependent movement strategy in which each agent switches between undirected and directed movement based on its perception of resources. This switching behavior is central to our goal of exploring how environmental and species-specific factors determine the evolution and maintenance of perception in an ecological system. We observed a non-linear response in the evolved perceptual ranges as a function of parameters in our model. Overall, we identified two groups of parameters, one of which promotes evolution of perception and another group that restricts it. We found that resource density, basal energy cost, perceptual cost and mutation rate were the best predictors of the resultant perceptual range distribution, but detailed exploration indicated that individual parameters affect different parts of the distribution in different ways. 

Abstract The role of transcription factors and biomolecules in cell type conversion has been widely studied. Yet, it remains unclear whether and how intracellular mechanotransduction through focal adhesions (FAs) and the cytoskeleton regulates the epigenetic state and cell reprogramming. Here, it is shown that cytoskeletal structures and the mechanical properties of cells are modulated during the early phase of induced neuronal (iN) reprogramming, with an increase in actin cytoskeleton assembly induced by Ascl1 transgene. The reduction of actin cytoskeletal tension or cell adhesion at the early phase of reprogramming suppresses the expression of mesenchymal genes, promotes a more open chromatin structure, and significantly enhances the efficiency of iN conversion. Specifically, reduction of intracellular tension or cell adhesion not only modulates global epigenetic marks, but also decreases DNA methylation and heterochromatin marks and increases euchromatin marks at the promoter of neuronal genes, thus enhancing the accessibility for gene activation. Finally, micro‐ and nano‐topographic surfaces that reduce cell adhesions enhance iN reprogramming. These novel findings suggest that the actin cytoskeleton and FAs play an important role in epigenetic regulation for cell fate determination, which may lead to novel engineering approaches for cell reprogramming.