Biology subjects

Hubert, M.

Publications and source records attributed to Hubert, M..

At least 19 recordsLinked to original sources

bioRxiv2026

A mitochondrial orphan protein contributes to sperm development in bivalves

Lineage-specific genes, or orphan genes, are increasingly recognized as important drivers of evolutionary innovation, yet their functional characterization remains largely restricted to nuclear genomes of model organisms. Whether lineage-specific genes encoded by mitochondrial DNA contribute to phenotypic diversification remains virtually unexplored. Here we investigate MORF, a rapidly evolving ORFan protein encoded by the paternally transmitted mitochondrial genome of bivalves with doubly uniparental inheritance (DUI). Combining immunolocalization, functional neutralization, protein interaction assays and proteomics in the distantly related bivalves Mytilus edulis and Venustaconcha ellipsiformis, we characterize MORF localization, function and molecular associations during male germ cell development. MORF localizes to developing sperm cells and mature sperm mitochondria in both species and is additionally detected in the acrosome of M. edulis. In cultured M. edulis germ cells, antibody-mediated neutralization of MORF increases the frequency of spermatids displaying four rather than the canonical five mitochondria, providing functional evidence that MORF contributes to mitochondrial organization during spermatogenesis. Comparative proteomic analyses identify MORF-associated proteins involved in cytoskeletal organization, protein homeostasis and extracellular matrix organization. The association between MORF and laminin is independently supported by western blotting in both species. Despite extensive sequence divergence, the two MORF proteins display overlapping functional and molecular features. Together, these findings uncover a functional role for a rapidly evolving mitochondrial orphan protein and point to mitochondrial orphan genes as a potentially overlooked source of evolutionary novelty.

molecular biology↗

bioRxiv2026

Apical-Out Ileal Organoids: A Versatile Platform for Probing Gut-Nanomaterial Interactions

The human ileal epithelium forms a critical interface governing interactions with luminal microorganisms and nanomaterials, yet physiologically relevant in vitro models with accessible apical surfaces remain limited. Here, human apical-out ileoids derived from healthy ileal biopsies are established as a platform that recapitulates the architecture and complexity of the native ileal epithelium. Assessment of donor- and passage-dependent variability identifies a representative ileoid line and a robust culture window. Integrated proteomic, imaging, ultrastructural, and functional analyses demonstrate preservation of key epithelial features, including brush-border organization, barrier integrity, absorptive capacity, and multiple endocytic pathways. Intact ileoids internalize nanoparticles of distinct surface charge, including empty lipid nanoparticles compositionally matched to approved mRNA vaccine formulations, establishing that uptake can be quantified across an unperturbed apical surface. This uptake is restrained by a retained mucin-rich glycocalyx. Cleavage of surface-attached mucins selectively enhances uptake of negatively charged nanoparticles, leaving near-neutral and positively charged nanoparticles unchanged, and increasing entry of Salmonella bacteria. These findings reveal the glycocalyx as a charge-selective regulator of epithelial uptake. In conclusion, our study provides a foundation for nanoparticle-epithelium investigations using human apical-out ileoids, and a versatile, translationally relevant platform for nanomedicine delivery, host-microbe interactions, and epithelial transport.

cell biology↗

bioRxiv2026

Viral rewiring of APC/C-CDC20 drives Aurora B hyperubiquitination, mitotic regression and polyploidy

Cytokinesis requires elaborate processes in the midbody, including the timely inactivation of Aurora B kinase (AurB) and membrane fission to separate the daughter cells. Here, we show that human adenovirus (AdV) perturbs midbody function, prevents daughter separation and induces cytoplasmic regression of both cancer cells and primary diploid human airway basal cells resulting in polyploidy. Infected cells undergo cleavage furrow regression after midbody formation driven by the viral protein E4orf4 independently of E4orf4 interaction with protein phosphatase 2A. The co-activator of the anaphase-promoting complex/cyclosome (APC/C), CDC20 is upregulated in AdV infection and required for regression. E4orf4 directly interacts with CDC20 and redirects the E3-ubiquitin ligase APC/C-CDC20 towards AurB, promoting hyperubiquitination and premature extraction of AurB from the midbody. Loss of midbody-associated AurB coincides with midbody collapse and furrow regression. Our findings reveal a previously unrecognized APC/C-CDC20-dependent pathway controlling late cytokinesis through AurB extraction from the midbody.

cell biology↗

bioRxiv2026

Human airway organoids as a high-throughput screening platform for antiviral natural products discovery

Antiviral drug discovery for respiratory viruses is hindered by the lack of scalable physiologically relevant systems. Here, we report the first high-throughput screen of 764 natural plant extracts against respiratory syncytial virus (RSV) using human primary airway organoids as a relevant model. A parallel screen conducted in A549 cells allowed the identification of 70 extracts with organoid-specific antiviral activity from which 45 active phytocompounds were purified. We identified early- and late-acting antiviral compounds and demonstrated a polarization-dependent activity for some of them. Collectively, our results establish the use of airway organoids as a scalable first-line platform for high-throughput antiviral discovery and exploit the plant-derived chemical space as an underexplored source of RSV inhibitors.

microbiology↗

bioRxiv2025

Generation of apical-out nasal organoids to facilitate viral infection and drug screening

Advanced culture systems such as organoids can serve as powerful platforms to study epithelial physiology, as they recapitulate the organisation and many key functions of the tissue of origin. The nasal epithelium is the first respiratory epithelium that is exposed to inhaled airborne pathogens. As a result, it is crucial to model host-pathogen interactions occurring in this tissue. To facilitate the efficient modelling of these interactions, we have developed a method to generate de novo apical-out nasal organoids from nasal epithelial cell aggregates. Optimisation of this method revealed a stark tissue-specific effect of the culture temperature, as apical-out nasal organoids were generated in much higher efficiency at 32.5 {degrees}C, compared to more widely used temperatures of 37{degrees}C. These organoids are composed of ciliated, basal and goblet cells and are produced in a completely standardised and scalable manner, devoid of any extracellular matrix hydrogel. Moreover, they displayed high homogeneity in size and cellular composition, as well as susceptibility to viral infections and capability to model antiviral drug responses. Here, we describe a method for the efficient and reproducible generation of apical-out nasal organoids with high potential to be utilised in host-pathogen interaction studies and personalised medicine from easy-to-access nasal swabs.

cell biology↗

bioRxiv2025

Potent Neutralization by Antibodies Targeting the Mpox A28 Protein

Mpox is the most pathogenic Poxvirus in circulation. While several antigens have been identified as targets for neutralizing antibodies, many proteins remain unexplored. We isolated and characterized four monoclonal antibodies (mAbs) targeting the Mpox A28 (OPG153), a virulence factor present on mature Mpox virions. The antibodies were isolated from convalescent individuals, alongside 14 additional mAbs targeting the A35 and H3 proteins. Anti-A28 mAbs potently neutralized Mpox and Vaccinia virus (VACV) through complement-dependent mechanisms involving C1q and C3 deposition. High resolution crystal structures of Anti-A28 mAbs 10M2146 and 8M2110 in complex with VACV A26 revealed two proximal epitopes within the N-terminal domain. Passive transfer of 8M2110 attenuated disease in infected mice. Moreover, immunization with A28 elicited antigen-specific B cells and robust neutralizing antibody responses and provided complete protection against lethal VACV challenge. These findings support Mpox A28 as a promising target for the induction of neutralizing antibodies and antiviral interventions.

immunology↗

bioRxiv2025

Erythrocytosis-inducing PHD2 mutations implicate biological role for N-terminal prolyl-hydroxylation in HIF1α oxygen-dependent degradation domain

Mutations in EGLN1, the gene encoding for hypoxia-inducible factor (HIF) prolyl-4-hydroxylase 2 (PHD2), cause erythrocytosis and in rare cases the development of neuroendocrine tumors. In the presence of oxygen, PHD2 hydroxylates one or both conserved prolines in the oxygen-dependent degradation domain (ODD) of HIF subunits, sufficiently marking HIF for binding and ubiquitylation via the von Hippel-Lindau (VHL) tumor suppressor protein-containing E3 ubiquitin ligase and subsequent degradation by the 26S proteasome. However, prolyl-hydroxylation in the C-terminal ODD appears to be the predominant and sufficient event in triggering the oxygen-dependent destruction of HIF, rendering the biological significance of N-terminal ODD proline unclear. Here, we examined 7 disease-associated EGLN1 mutations scattered across the catalytic core and show definitively that all PHD2 mutants have a structural and/or catalytic activity defect as measured by time-resolved nuclear magnetic resonance. Notably, we identified one of the PHD2 mutants, P317R, to retain comparably wild-type capacity to hydroxylate the predominant proline in the C-terminal ODD but had uniquely compromised ability to hydroxylate the N-terminal ODD proline. These findings support the notion that deregulation of HIF ultimately underlies PHD2-driven erythrocytosis and challenge the currently held uncertainty that the N-terminal ODD prolyl-hydroxylation event is dispensable in normal hypoxic signaling pathway.

biochemistry↗

bioRxiv2025

IntegrAlign: A comprehensive tool for multiimmunofluorescence panel integration through image alignment

MotivationTyramide-based multiplex-immunofluorescence (mIF) enables the simultaneous analysis of up to seven protein markers on a whole slide, providing a comprehensive approach to study the tumor microenvironment. Integrating multiple mIF panels through image alignment of serial slide significantly expands the number of cell populations analyzed in a single space. IntegrAlign was developed to optimize this integration on serial whole slides, enhancing the value and applicability of mIF for comprehensive spatial analyses and enabling biomarker discovery at scale. ResultsIntegrAlign, leveraging the SimpleITK toolkit, applies a two-step alignment using rigid and B-spline transformations to integrate serial mIF whole slides. Validation on simulated and real datasets demonstrated alignment accuracy below the diameter of a cell nucleus ([~]6 {micro}m), outperforming existing methods. This precision enhances spatial analyses by combining extended phenotypic data, supporting novel insights into tissue architecture and cellular interactions. Availability and ImplementationIntegrAlign is open-source, implemented in Python, and freely available under the MIT license at https://github.com/CAUXlab/IntegrAlign.

bioinformatics↗

bioRxiv2025

Human jejunal enteroids for studies of epithelial drug transport and metabolism

Intestinal enteroids are stem cell-based "mini-guts" that mimic many aspects of the corresponding epithelial barrier in vivo. Here, we established and characterized differentiated apical-out (AO) and basal-out (BO) jejunal enteroids in suspension and followed their differentiation by quantitative global proteomics and different microscopic techniques. The barrier integrity and function and subcellular location of nutrient and clinically important drug transporters were investigated in the matured enteroids using live-cell microscopy. The presystemic metabolism of two drugs by CYP3A4 was determined and the results were used to predict the pharmacokinetics after oral administration by a PBPK population model. The differentiated AO enteroids displayed a protein profile that overlapped both qualitatively and quantitatively with that of freshly isolated jejunal enterocytes and tissue. They exhibited a morphology that recapitulates the mature villus enterocyte in vivo, formed an intact barrier with a well-developed glycocalyx and are impermeable to the hydrophilic low molecular weight compound lucifer yellow and transported a medium chain fatty acid derivative by FATP4 into lipid deposits. The clinically important ABC-transporters Pgp and BCRP were expressed at near in vivo levels, had the correct subcellular localization and effluxed their substrates. Terfenadine and midazolam were metabolized by CYP3A4 and the results were used to predict the clinical pharmacokinetics of the drugs after oral administration with good accuracy. We conclude that suspended 3D AO enteroids provide a physiologically relevant model for studies of intestinal function that offers convenient access to the apical surface and is easy to dispense in multi-wells formats for large scale experimentation.

cell biology↗

bioRxiv2025

Oncogenic Stress is a Novel Immunogenic Signal Driven by the Unfolded Protein Response and Detected by Neutrophils

Breast cancer (BC) is the leading cause of cancer-related death in women. However, early detection of BC remains a major clinical challenge and represents a significant obstacle to effective prevention. To improve early clinical management, a deeper understanding of the preneoplastic immune microenvironment of BC is crucial. Among innate immune populations, neutrophils have emerged as important modulators of tumor development, but their role during the initiation of BC remains poorly understood. By integrating depletion experiments with transcriptomic profiling of sorted preneoplatic epithelial cells and neutrophils in spontaneous breast cancer mouse models, we observed that neutrophils contribute to tumor surveillance of preneoplastic stage with the activation of the unfolded protein response (UPR) in the preneoplastic epithelial compartment. To decipher the early anti-tumoral role of neutrophil, we developed an in vitro co-culture model of human mammary epithelial cells undergoing oncogenic stress with activation of the UPR (eHMEC), with human primary neutrophils. eHMEC display an immunoactive secretome as well as immunogenic membrane ligands, and neutrophils are the only immune cell population detecting eHMEC immunogenic signals leading to their recruitment, activation, production of reactive oxygen species and degranulation. Altogether, our work identifies for the first-time neutrophils as the earliest immune cell involved in immunosurveillance of preneoplastic BC epithelial cells, paving the way for potential therapeutic approaches targeting neutrophils to intercept early steps of BC tumorigenesis.

immunology↗

bioRxiv2024

Spatial architecture of CD8+ T cells and DC subsets is critical for the response to immune checkpoint inhibitors in melanoma

BackgroundDendritic cells (DCs) are promising targets for cancer immunotherapies owing to their central role in the initiation and the control of immune responses. Their functions encompass a wide range of mechanisms mediated by different DC subsets. Several studies have identified human tumor- associated DC (TA-DC) populations through limited marker-based technologies, such as immunostaining or flow cytometry. However, tumor infiltration, spatial organization and specific functions in response to immunotherapy of each DC subset remain to be defined. MethodsHere, we implemented a multiplexed immunofluorescence analysis pipeline coupled with bio-informatic analyses to decipher the tumor DC landscape and its spatial organization within melanoma patients lesions, and its association with patients response to immune checkpoint inhibitors (ICI). For this aim, we analyze a cohort of 41 advanced melanoma patients treated with anti- PD1 alone or associated with anti-CTLA4. Distance and cell network analyses were performed to gain further insight into the spatial organization of tumor-associated DCs. A Digital Spatial Profiling analysis further characterized ecosystem of tumor-infiltrating DCs. ResultsPlasmacytoid DCs (pDCs) were the most abundant DC population, followed by conventional cDC1 and mature DCs, present in equal proportions. In contrast to CD8+ T cell frequency, and despite varying densities, all DC subsets were associated with a favorable response to ICI. Distance and cell network analyses demonstrated that tumor-infiltrating DCs were largely organized in dense areas with high homotypic connections, except for cDC1 that exhibited a more scattered distribution. We identified four patterns of ecosystems with distinct preferential interactions between DC subsets. Significantly, the proximity and interactions between CD8+ T cells and cDC1 were positively associated with patients response to ICI. ConclusionsOur study unravels the complex spatial organization of DC subsets and their interactions in melanoma patient lesions, shedding light on their pivotal role in shaping the response to ICI. Our discoveries regarding the spatial arrangement of cDC1, especially with CD8+ T cells, provide valuable clues for improving immunotherapeutic strategies in melanoma patients. What is already known on this topicDendritic cells (DCs) are promising targets for cancer immunotherapies owing to their central role in the initiation and the control of immune responses. Although conventional type 1 dendritic cells (cDC1) were proposed to contribute to immunotherapy response, their precise functions and interactions with other immune populations in human cancers are largely unknown. What this study addsThis study provides a precise characterization of the spatial distribution and organization of tumor- infiltrating DCs in a large cohort of advanced melanoma patients, and in correlation with response to immunotherapy. While DCs are organized in dense areas with high homotypic connections, cDC1 exhibit a more scattered distribution and form heterotypic aggregates with other DC subsets. More importantly, a close connection between cDC1 and CD8 T cell is uniquely correlated with the patients response to immunotherapy. How this study might affect research, practice or policyThis study improves our understanding of CD8-DC spatial organization within the tumor microenvironment and will have a broad spectrum of implications in the design of anti-tumor immune-activating compounds and the design of biomarkers of response to immunotherapy for melanoma patients.

cancer biology↗

bioRxiv2023

Distinct evolution of SARS-CoV-2 Omicron XBB and BA.2.86 lineages combining increased fitness and antibody evasion

The unceasing circulation of SARS-CoV-2 leads to the continuous emergence of novel viral sublineages. Here, we isolated and characterized XBB.1, XBB.1.5, XBB.1.9.1, XBB.1.16.1, EG.5.1.1, EG.5.1.3, XBF, BA.2.86.1 and JN.1 variants, representing >80% of circulating variants in January 2024. The XBB subvariants carry few but recurrent mutations in the spike, whereas BA.2.86.1 and JN.1 harbor >30 additional changes. These variants replicated in IGROV-1 but no longer in Vero E6 and were not markedly fusogenic. They potently infected nasal epithelial cells, with EG.5.1.3 exhibiting the highest fitness. Antivirals remained active. Neutralizing antibody (NAb) responses from vaccinees and BA.1/BA.2-infected individuals were markedly lower compared to BA.1, without major differences between variants. An XBB breakthrough infection enhanced NAb responses against both XBB and BA.2.86 variants. JN.1 displayed lower affinity to ACE2 and higher immune evasion properties compared to BA.2.86.1. Thus, while distinct, the evolutionary trajectory of these variants combines increased fitness and antibody evasion.

microbiology↗

bioRxiv2023

High fusion and cytopathy of SARS-CoV-2 variant B.1.640.1

SARS-CoV-2 variants with undetermined properties have emerged intermittently throughout the COVID-19 pandemic. Some variants possess unique phenotypes and mutations which allow further characterization of viral evolution and spike functions. Around 1100 cases of the B.1.640.1 variant were reported in Africa and Europe between 2021 and 2022, before the expansion of Omicron. Here, we analyzed the biological properties of a B.1.640.1 isolate and its spike. Compared to the ancestral spike, B.1.640.1 carried 14 amino acid substitutions and deletions. B.1.640.1 escaped binding by some anti-NTD and -RBD monoclonal antibodies, and neutralization by sera from convalescent and vaccinated individuals. In cell lines, infection generated large syncytia and a high cytopathic effect. In primary airway cells, B.1.640.1 replicated less than Omicron BA.1 and triggered more syncytia and cell death than other variants. The B.1.640.1 spike was highly fusogenic when expressed alone. This was mediated by two poorly characterized and infrequent mutations located in the spike S2 domain, T859N and D936H. Altogether, our results highlight the cytopathy of a hyper-fusogenic SARS-CoV-2 variant, supplanted upon the emergence of Omicron BA.1. ImportanceOur results highlight the plasticity of SARS-CoV-2 spike to generate highly fusogenic and cytopathic strains with the causative mutations being uncharacterized in previous variants. We describe mechanisms regulating the formation of syncytia and the subsequent consequences in cell lines and a primary culture model, which are poorly understood.

microbiology↗

bioRxiv2023

Laminin alpha 5 - induced mechanical homeostasis modulates retinal epithelium functionality

Epithelial cells are highly interconnected, whereby they acquire mesoscale mechanical properties to accomplish specific tissue functions. In homeostasis, this mechanical status can be summarised as mechanical homeostasis, regulated by the balance of intercellular tension and extracellular matrix adhesion forces. In the outer retina, the significance of this force balance and its consequences for vision remains poorly understood. We found that the density of basement membrane laminins modulates the level of retinal pigmented epithelium contractility, which directly controls its efficiency in phagocytosing photoreceptor outer segments. In vivo, the density gradient of laminins follows retinal functional demand, supporting the physiological role of laminins in controlling epithelial mechanical homeostasis. Our data suggest that laminin density and isoform heterogeneity can differentially engage integrins {beta}1 and {beta}4, the ratio of which determines the contribution of actin vs keratin cytoskeleton in balancing tissue mechanics. With this work, we suggest that the extracellular matrix-defined mechanical status of retinal pigmented epithelium is a novel parameter for visual function. SignificanceIn the retina, the retinal pigmented epithelium (RPE) is responsible for the daily phagocytosis of photoreceptor cell fragments, a process vital for visual function. Along the visual axis, there is a natural decrease in the ratio of photoreceptors to RPE cells, indicating a decrease in RPE functional demand. This study reveals that the density of laminins in RPE basement membrane also diminishes along this axis, critically influencing RPE function by regulating its contractility. For the first time, we demonstrate the presence of a laminin- defined mechanical gradient within the RPE, which determines its capacity to support photoreceptor cells. Our findings highlight the importance of mechanical properties as a key factor in visual function, offering new insights into retinal health and disease.

biophysics↗

bioRxiv2023

IL-33 drives polyfunctionality and antitumor activity of a unique ST2+ NK cell population

Despite increasing recognition of NK cell diversity, their functional heterogeneity and its relevance to cancer remain incompletely understood. Here, we show that IL-12 promotes the emergence of a distinct ST2/IL33R+ NK cell state poised for robust activation by IL-33. Human ST2 NK cells exhibit a unique transcriptional program, an intermediate differentiation state between CD56bright and CD56dim NK cells, and enhanced polyfunctionality combining proliferation, cytokine production, and cytotoxicity. ST2 NK cells were identified across human and murine tumor datasets. In murine breast cancer models, IL-33 and IL-12 cooperated to elicit potent antitumor responses dependent on IFN-{gamma} and ST2 NK cells. In cancer patients, IL-12 unleashed tumor-infiltrating ST2 NK cells' potential to produce IFN-g in response to IL-33, an IL33hi-NKhi score was associated with prolonged survival, and ST2 NK cells correlated with improved immunotherapy response. Collectively, these findings identify ST2 NK cells as a therapeutically actionable NK cell state promoting antitumor immunity.

cancer biology↗

bioRxiv2023

CD4 T cells and neutrophils contribute to epithelial-mesenchymal transition in breast cancer

Epithelial-mesenchymal transition (EMT) is a central oncogenic mechanism, contributing both to transformation and metastatic dissemination. Inflammation and innate immune cells are known to favor EMT induction, but the role of adaptive immunity still remains unclear. Using an original murine mammary tumor model in immune cell subpopulation depletion experiments, we demonstrated that tumor cells maintain their epithelial phenotype in mice deficient for adaptive immune response, but undergo EMT in the presence of T-cells. This phenotypic conversion involves the major contribution of CD4 T cells, but not CD8 T cells nor B cells, undoubtedly demonstrating the pro-EMT role of CD4 T cells specifically among adaptive immune cells. Moreover, combined intra-tumor immune infiltrate and transcriptomic analyses of murine mammary tumors with various EMT phenotype revealed an inverse correlation between mesenchymal tumor cell and intratumoral neutrophil proportions, due to the reduced ability of mesenchymal cells to recruit neutrophils. Last, selective in vivo depletion of neutrophils and transcriptomic analysis of human breast tumor cohorts demonstrated the pro-EMT role of neutrophils and suggest a cooperation with CD4 T cells in EMT promotion. Collectively, our data highlight a novel mechanism of EMT regulation by both innate and adaptive immune compartments.

cancer biology↗

bioRxiv2023

Capturing the mechanosensitivity of cell proliferation in models of epithelium

Despite the primary role of cell proliferation in tissue development and homeostatic maintenance, the interplay between cell density, cell mechanoresponse, and cell growth and division is not yet understood. In this article we address this issue by reporting on an experimental investigation of cell proliferation on all time- and length-scales of the development of a model tissue, grown on collagen-coated glass or deformable substrates. Through extensive data analysis, we demonstrate the relation between mechanoresponse and probability for cell division, as a function of the local cell density. Motivated by these results, we construct a minimal model of cell proliferation that can recover the data. By parametrizing the growth and the dividing phases of the cell cycle, and introducing such a proliferation model in dissipative particle dynamics simulations, we recover the mechanoresponsive, time-dependent density profiles in 2D tissues growing to macroscopic scales. The importance of separating the cell population into growing and dividing cells, each characterized by a particular time scale, is further emphasized by calculations of density profiles based on adapted Fisher-Kolmogorov equations. Together, these results show that the mechanoresponse on the level of a constitutive cell and its proliferation results in a matrix-sensitive active pressure. The latter evokes massive cooperative displacement of cells in the invading tissue and is a key factor for developing large-scale structures in the steady state.

biophysics↗

bioRxiv2023

Morphology as indicator of adaptive changes of model tissues in osmotically and chemically changing environments

We investigate the formation and maintenance of the homeostatic state in the case of 2D epithelial tissues following an induction of hyperosmotic conditions, using media enriched with 80 to 320 mOsm of mannitol, NaCl, and urea. We characterise the changes in the tissue immediately after the osmotic shock, and follow it until the new homeostatic state is formed. We characterise changes in cooperative motility and proliferation pressure in the tissue upon treatment with the help of a theoretical model based on the delayed Fisher-Kolmogorov formalism, where the delay in density evolution is induced by the the finite time of the cell division. Finally we explore the adaptation of the homeostatic tissue to highly elevated osmotic conditions by evaluating the morphology and topology of cells after 20 days in incubation. We find that hyperosmotic environments together with changes in the extracellular matrix induce different mechanical states in viable tissues, where only some remain functional. The perspective is a relation between tissue topology and function, which could be explored beyond the scope of this manuscript. O_LIExperimental investigation of morphological effect of change of osmotic conditions on long-term tissue morphology and topology C_LIO_LIEffect of osmotic changes on transient tissue growth behaviour C_LIO_LIAnalysis of recovery process of tissues post-osmotic-shock C_LIO_LIToxicity limits of osmolytes in mid- to long-term tissue evolution C_LIO_LITissue adaptation to physiological changes in environment C_LIO_LILong-term tissue stabilisation under altered osmotic conditions. C_LI O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=77 SRC="FIGDIR/small/524301v1_ufig1.gif" ALT="Figure 1"> View larger version (40K): org.highwire.dtl.DTLVardef@826d28org.highwire.dtl.DTLVardef@cc7f35org.highwire.dtl.DTLVardef@1b370fborg.highwire.dtl.DTLVardef@1bea90e_HPS_FORMAT_FIGEXP M_FIG C_FIG

biophysics↗