We present a constitutive model for the passive and active response of skeletal muscles. At variance with more classical approaches, the model is developed exploiting adapted Voigt representations of strain and stress tensors within the context of nonlinear Cauchy elasticity. This framework allows us to identify non-trivial stress-strain relations in a rather direct way from experimental data, enhancing the mechanical interpretability of the material functions that describe the tissue response and obtaining additional insight on the distinct role of the contractile fibres and of the surrounding extracellular matrix. We propose a two-material model, with an additive splitting of the stress contributions, in which only one component depends on an activation parameter. The constitutive model for the passive behaviour satisfactorily predicts the nonlinear stress response to elongation at different relative orientations with respect to the fibre direction and highlights the dominant role of the extracellular matrix. The activation model, essentially determined by the mechanics of the contractile fibres, captures well the isometric stress response through the prescription of an elasto-plastic evolution of the along-fibre active strain.
Congenital heart defects frequently arise from alterations in the elongation of the cardiac outflow tract (OFT). Proper elongation of the OFT depends on the coordinated deployment of progenitor cells from the second heart field (SHF) and on dynamic interactions with the extracellular matrix (ECM). Among ECM components, fibronectin (Fn1) and tenascin-C (TnC) have emerged as key regulators of cardiac morphogenesis. Studies in mouse embryos have shown that mesodermal Fn1 is required to maintain proper TnC localization within SHF cells. To study heart development, mammalian models are challenging to use because of their in utero development. This limitation highlights the need for alternative models with external development, where direct observation is possible; however, in these systems, the cellular organization of the SHF and the dynamics of its ECM environment remain poorly characterized Here, we investigated the cellular and extracellular architecture of SHF cells localized to the dorsal pericardial wall (DPW) during heart development in Xenopus laevis. We show that SHF cells undergo a stage-dependent transition from a predominantly monolayered organization at NF35 to a multilayered structure at NF42. This transition is accompanied by dynamic remodeling of the ECM, characterized by increased expression of Fn1, TnC, and Collagen I (ColI) and by redistribution of ECM components within the DPW. Functional experiments revealed that depletion of Fn1 disrupts cardiac morphogenesis, leading to shortening of the OFT and reduced ventricular size. Moreover, loss of Fn1 decreases TnC and ColI levels and alters the spatial organization of TnC within the DPW, indicating that Fn1 is required for proper ECM assembly within the SHF cells. These findings identify Fn1 as a key regulator of ECM assembly within the DPW and highlight how ECM remodeling contributes to the organization of SHF progenitor cells during OFT elongation. Altogether, we demonstrated that Xenopus laevis is a powerful model for studying ECM-driven mechanisms of cardiac morphogenesis. ### Competing Interest Statement The authors have declared no competing interest. Fondecyt Iniciacion, 11240544, 11220624 IBRO Centro Ciencia & Vida, FB210008
Coral white syndrome (WS) is a widespread condition characterized by tissue loss and skeletal exposure, with multiple bacterial pathogens, particularly Vibrio species, implicated in its etiology. To investigate the genomic basis of potential virulence in Vibrio associated with WS of Porites cylindrica, we isolated 57 Vibrio strains from healthy and diseased coral tissues collected from reefs in Guam. Thirteen representative strains from five dominant species, V. coralliilyticus, V. owensii, V. tubiashii, V. harveyi, and V. tetraodonis, were selected for whole-genome sequencing using Oxford Nanopore Technology. Comparative genomic analyses revealed a conserved repertoire of extracellular enzymes, including hemolysins, cytolysins, metalloproteases, and subtilisin-like peptidases, alongside lineage-specific toxin and regulatory modules. Variation in secretion systems (T1SS β T6SS), particularly in T3SS, T4SS, and T6SS subtypes, reflected diversification of host-interaction and competitive capacities across species. Mobile genetic elements, including plasmids and prophages, contributed additional virulence-associated genes and secretion clusters, underscoring the role of horizontal gene transfer in shaping the accessory genome content. Notably, V. coralliilyticus harbored cholera toxin related genes (ace and zot), while conjugative plasmid systems indicated potential for gene dissemination across lineages. Together, these findings demonstrate that virulence potential in coral-associated Vibrio is broadly distributed and structured by a conserved ecological core overlaid with flexible, horizontally acquired modules. This study provides the first comparative genomic framework for Vibrio associated with P. cylindrica, advancing our understanding of how genomic plasticity and modular virulence repertoires may contribute to opportunistic disease dynamics in coral reef ecosystems. ### Competing Interest Statement The authors have declared no competing interest. U.S. National Science Foundation, OIA-1946352
Signalling via the epidermal growth factor receptor (EGFR) is indispensable for morphogenesis and tissue homeostasis. It is activated by extracellular ligands, typically released from transmembrane precursors by proteolysis. Ligand shedding activity is provided by the conserved rhomboid intramembrane serine proteases in Drosophila, but by the unrelated ADAM family metalloproteases in mammals, leaving the functions of mammalian non-mitochondrial rhomboids underexplored. Using quantitative proteomics, we show that EGFR is the main endogenous substrate of the human rhomboid protease RHBDL2 in keratinocytes. By shedding the EGFR ectodomain, thus producing a decoy receptor, RHBDL2 suppresses EGFR signalling, limiting cell migration and invasion. Conspicuously, RHBDL2 activity is upregulated by elevated intracellular calcium concentration, a condition typical for keratinocyte differentiation. These effects are recapitulated in primary human keratinocytes, and human skin equivalents deficient in RHBDL2 display incomplete differentiation and are morphologically disordered compared to wild type cells. We propose that context-specific fine-tuning of EGFR signalling and sensitivity to cross-talk from other signalling pathways could be important and hitherto overlooked roles of rhomboid proteases in mammals.
Heart regeneration requires coordinated immune activation, timely inflammatory resolution, and dynamic extracellular matrix (ECM) remodeling in addition to cardiomyocyte (CM) proliferation. However, the cytokine signals that instruct immune cell functions during cardiac repair remain incompletely understood. Here, we identify interferon-gamma (IFN-Ξ³) as a critical regulator of macrophage plasticity in zebrafish heart regeneration. IFN-Ξ³ signaling components are dynamically activated following cardiac injury, with early induction of ifng1 and temporally coordinated receptor expression. Genetic ablation of ifng1 impairs myocardial regeneration, resulting in reduced CM proliferation and persistent fibrotic scarring. Temporal transcriptional profiling reveals sustained inflammatory signatures, impaired efferocytosis, and abolished reparative programs, accompanied by aberrant immune cell dynamics and retention of injury-derived debris in mutant hearts. Transcriptomic analysis of cardiac macrophages further reveals that IFN-Ξ³ deficiency disrupts the transition from an inflammatory state to a reparative, ECM-remodeling phenotype, leading to reduced collagen denaturation and diminished CM protrusion at the injury border zone. Inducible- and macrophage-specific blockade of IFN-Ξ³ signaling phenocopies defects in global knockout, establishing a cell-autonomous requirement for IFN-Ξ³ in coordinating regenerative immune function. Collectively, our findings define an IFN-Ξ³-dependent macrophage reprogramming axis that couples inflammatory resolution to ECM remodeling in heart regeneration, elucidating how cytokine signaling actively instructs tissue repair. ### Competing Interest Statement The authors have declared no competing interest. National Science and Technology Council, NSTC 114-2320-B-001-010-MY3, NSTC 113- 548 2923-B-001-003-MY2 Academia Sinica, https://ror.org/05bxb3784, AS-CDA-112-L04, AS-GC- 551 110-05 Institute of Biomedical Sciences, Academia Sinica, IBMS-CRC108-P01, IBMS-CRC111- 550 P01
Fibrosis involves sustained changes in fibroblast gene expression, leading to excessive extracellular matrix (ECM) deposition and progressive tissue stiffening. Although matrix stiffness is a potent regulator of cell fate and transcription, how nuclear mechanosensing contributes to fibrosis remains unclear. Here, we define a central role for SUN2, a component of linker of nucleoskeleton and cytoskeleton (LINC) complexes, as a mediator of stiffness-dependent nuclear and chromatin responses during skin fibrosis. SUN2 transcripts are upregulated in dermal fibroblasts of patients with systemic sclerosis and Sun2 protein is elevated in fibrotic mouse skin. Nuclear size, A-type lamins and Sun2 are elevated in dermal fibroblasts plated on stiff substrates. Loss of Sun2 protects against bleomycin-induced skin fibrosis in vivo and abolishes stiffness-induced changes in nuclear size and fibrotic gene expression in vitro. Mechanistically, we identify three Sun2-dependent mechanosensitive chromatin states and show that mechanical induction of the histone methyltransferase Ezh2 requires Sun2. These findings define SUN2 as a nuclear mechanosensor that couples matrix stiffness to chromatin regulation and transcriptional programs that drive fibrosis, identifying it as a potential therapeutic target pathway in fibrotic disease. ### Competing Interest Statement The authors receive research funding from Boehringer-Ingelheim Pharmaceutical, Inc. National Institute of Arthritis and Musculoskeletal and Skin Diseases, https://ror.org/006zn3t30, AR076938, AR0695505, AR084558, AR085488 National Institute of General Medical Sciences, https://ror.org/04q48ey07, GM153474 LEO Foundation, https://ror.org/02rgsr590 Boehringer Ingelheim (Germany), https://ror.org/00q32j219
Fibrosis and pathological stiffening of tissue are driven by mechanical and biochemical signaling pathways. Here, we find that Sun2, an integral inner nuclear membrane component of Linker of Nucleoskeleton and Cytoskeleton (LINC) complexes, is up-regulated in the lung of patients suffering from fibrotic conditions and in fibroblasts during an injury-induced mouse model of lung fibrosis. Sun2 protein levels also increase in primary lung fibroblasts in a substrate stiffness-dependent manner. Sun2-/- primary lung fibroblasts respond to TGFΞ², become contractile, and express a key marker of extracellular matrix-producing fibroblasts, Cthrc1 . Consistent with this, Sun2 is dispensable for myofibroblast formation and repairing the alveolar barrier after bleomycin injury. Remarkably, however, fibrosis does not develop in bleomycin-treated Sun2-/- mouse lungs. This is explained by the requirement for Sun2 to up-regulate genes encoding extracellular matrix proteins. We therefore suggest that Sun2-containing LINC complexes contribute to a mechanical coincidence detection mechanism that acts in concert with canonical TGFΞ² signaling necessary for pathologic extracellular matrix protein production, representing a nuclear mechanosensing node for intervention in fibrotic diseases of the lung. ### Competing Interest Statement The authors have declared no competing interest. National Institutes of Health, F31HL158119, F31AR085488, R35GM153474, 5R01HL163984, 1R01HL178097-01A1, R01AR076938 National Institutes of Health, https://ror.org/01cwqze88, R01AR0695505, R01AR084558
Rxiv mechanobio