Mechanical stress confers nuclear and functional changes in derived leukemia cells from persistent confined migration.

Nuclear deformability plays a critical role in cell migration. During this process, the remodeling of internal components of the nucleus has a direct impact on DNA damage and cell behavior; however, how persistent migration promotes nuclear changes leading to phenotypical and functional consequences remains poorly understood. Here, we described that the persistent migration through physical barriers was sufficient to promote permanent modifications in migratory-altered cells. We found that derived cells from confined migration showed changes in lamin B1 localization, cell morphology and transcription. Further analysis confirmed that migratory-altered cells showed functional differences in DNA repair, cell response to chemotherapy and cell migration in vivo homing experiments. Experimental modulation of actin polymerization affected the redistribution of lamin B1, and the basal levels of DNA damage in migratory-altered cells. Finally, since major nuclear changes were present in migratory-altered cells, we applied a multidisciplinary biochemical and biophysical approach to identify that confined conditions promoted a different biomechanical response of the nucleus in migratory-altered cells. Our observations suggest that mechanical compression during persistent cell migration has a role in stable nuclear and genomic alterations that might handle the genetic instability and cellular heterogeneity in aging diseases and cancer.

Moulding hydrodynamic 2D-crystals upon parametric Faraday waves in shear-functionalized water surfaces.

Faraday waves, or surface waves oscillating at half of the natural frequency when a liquid is vertically vibrated, are archetypes of ordering transitions on liquid surfaces. Although unbounded Faraday waves patterns sustained upon bulk frictional stresses have been reported in highly viscous fluids, the role of surface rigidity has not been investigated so far. Here, we demonstrate that dynamically frozen Faraday waves-that we call 2D-hydrodynamic crystals-do appear as ordered patterns of nonlinear gravity-capillary modes in water surfaces functionalized with soluble (bio)surfactants endowing in-plane shear stiffness. The phase coherence in conjunction with the increased surface rigidity bears the Faraday waves ordering transition, upon which the hydrodynamic crystals were reversibly molded under parametric control of their degree of order, unit cell size and symmetry. The hydrodynamic crystals here discovered could be exploited in touchless strategies of soft matter and biological scaffolding ameliorated under external control of Faraday waves coherence.

Multiple particle tracking analysis in isolated nuclei reveals the mechanical phenotype of leukemia cells.

The nucleus is fundamentally composed by lamina and nuclear membranes that enclose the chromatin, nucleoskeletal components and suspending nucleoplasm. The functional connections of this network integrate external stimuli into cell signals, including physical forces to mechanical responses of the nucleus. Canonically, the morphological characteristics of the nucleus, as shape and size, have served for pathologists to stratify and diagnose cancer patients; however, novel biophysical techniques must exploit physical parameters to improve cancer diagnosis. By using multiple particle tracking (MPT) technique on chromatin granules, we designed a SURF (Speeded Up Robust Features)-based algorithm to study the mechanical properties of isolated nuclei and in living cells. We have determined the apparent shear stiffness, viscosity and optical density of the nucleus, and how the chromatin structure influences on these biophysical values. Moreover, we used our MPT-SURF analysis to study the apparent mechanical properties of isolated nuclei from patients of acute lymphoblastic leukemia. We found that leukemia cells exhibited mechanical differences compared to normal lymphocytes. Interestingly, isolated nuclei from high-risk leukemia cells showed increased viscosity than their counterparts from normal lymphocytes, whilst nuclei from relapsed-patient's cells presented higher density than those from normal lymphocytes or standard- and high-risk leukemia cells. Taken together, here we presented how MPT-SURF analysis of nuclear chromatin granules defines nuclear mechanical phenotypic features, which might be clinically relevant.

A mesoscopic theory to describe the flexibility regulation in F-actin networks: An approach of phase transitions with nonlinear elasticity.

The synthetic actin network arouses great interest as bio-material due to its soft and wet nature that mimics many biological scaffolding structures. Inside the cell, the actin network is regulated by tens of actin-binding proteins (ABP's), which make for a highly complex system with several emergent behaviors. In particular, calponin is an ABP that was identified as an actin stabiliser, but whose mechanism is still poorly understood. Recent experiments using an in vitro model system of cross-linked actin with calponin and large deformation bulk rheology, found that networks with exhibited a delayed onset and were able to withstand a higher maximal strain before softening. In this work, we show that at network scale the actin network with calponin furthermore the reduction of the persistence length allows: (i) The reduction in the network pre-strain. (ii) The increment of the crosslinks adhesion energy. We verify these effects theoretically using nonlinear continuum mechanics for the semiflexible and crosslinked network. In addition, the alterations over the microstructure are described by the definition of an interaction parameter Γ according the formalism of Landau for phase transitions. According to this model we demonstrates that the interaction parameter can describe the experimental observations following a scaling exponent as Γ~|c-ccr|1/2, where c is the ratio between concentration of calponin and actin. This result provides interesting feedback to improve our understanding of several mechano-biological pathways.

Towards the understanding of cytoskeleton fluidisation-solidification regulation.

The understanding of the self-regulation of the mechanical properties in non-sarcomeric cells, such as lung cells or cells during tissue development, remains an open research problem with many unresolved issues. Their behaviour is far from the image of the traditionally studied sarcomeric cells, since the crosstalk between the signalling pathways and the complexity of the mechanical properties creates an intriguing mechano-chemical coupling. In these situations, the inelastic effects dominate the cytoskeletal structure showing phenomena like fluidisation and subsequent solidification. Here, we proposes the inelastic contractile unit framework as an attempt to reconciles these effects. The model comprises a mechanical description of the nonlinear elasticity of the cytoskeleton incorporated into a continuum-mechanics framework using the eighth-chains model. In order to address the inelastic effect, we incorporate the dynamic of crosslinks, considering the [Formula: see text]-actinin and the active stress induced by the myosin molecular motors. Finally, we introduce a hypothesis that links the ability to fluidise and re-solidify as a consequence of the interaction between the active stress and the gelation state defined by the crosslinks. We validate the model with data obtained from experiments of drug-induced relaxation reported in the literature.

Mechanics of epithelial closure over non-adherent environments.

The closure of gaps within epithelia is crucial to maintain its integrity during biological processes such as wound healing and gastrulation. Depending on the distribution of extracellular matrix, gap closure occurs through assembly of multicellular actin-based contractile cables or protrusive activity of border cells into the gap. Here we show that the supracellular actomyosin contractility of cells near the gap edge exerts sufficient tension on the surrounding tissue to promote closure of non-adherent gaps. Using traction force microscopy, we observe that cell-generated forces on the substrate at the gap edge first point away from the centre of the gap and then increase in the radial direction pointing into the gap as closure proceeds. Combining with numerical simulations, we show that the increase in force relies less on localized purse-string contractility and more on large-scale remodelling of the suspended tissue around the gap. Our results provide a framework for understanding the assembly and the mechanics of cellular contractility at the tissue level.