Biology subjects

Ros, M.

Publications and source records attributed to Ros, M..

9 recordsLinked to original sources

bioRxiv2025

Establishment, conservation, and innovation of dorsal determination mechanisms during the evolution of vertebrate paired appendages

Limb function requires polarized anatomy across the dorsal-ventral (DV) axis, but it is unclear when the capacity for DV differentiation of paired appendages arose in evolution. Here we define ancestral DV patterning programs in the fins of fishes. We show that the orthologue of the limb dorsal determinant, Lmx1b, is required to establish dorsality in zebrafish pectoral fins and is regulated by a conserved LARM cis-regulatory hub. However, lmx1bb expression in median fins is unaffected by removal of the LARM, suggesting its regulation is an evolutionary innovation specific to the paired appendages. Although we find the LARM is highly conserved across gnathostomes, we identify specific alteration of this region in hillstream loaches, fishes which naturally parallel "double-ventral" fin phenotypes observed in lmx1bb and LARM mutants. Altogether our findings indicate LARM-mediated dorsal identity is an ancestral feature of paired appendages that provide a prepattern for limb evolution and lineage diversification.

evolutionary biology↗

bioRxiv2025

Cancer cells transfer invasive properties through microRNAs contained in collagen-tracks

Invasion is a prerequisite for metastasis formation. During tumor development, the extracellular matrix (ECM) is remodeled in part through overexpression of type I collagen, increasing tumor microenvironment stiffness, and facilitating cancer dissemination. During breast cancer cell migration, we observed membrane debris left behind, attached to the collagen fibrils, along the migration path. We named these structures collagen-tracks. These collagen-tracks can be deposited in 3D matrices in vitro and in vivo and their formation is stimulated by the interaction between the ECM and matrix receptors, such as the discoidin-domain receptor (DDR1). However, they are different from structures already known to be involved in cell-cell communication such as exosomes and migrasomes, due to their specific nucleic acid and protein contents. When deposited by highly invasive breast cancer cells, internalized collagen-tracks reprogram non-invasive cells into highly pro-metastatic ones by inducing a partial epithelial-mesenchymal transition (EMT). This cell reprogramming is dependent on specific miRNAs present in the collagen-tracks, that are necessary to promote ECM degradation, increase cell motility and invasiveness. Collagen-tracks thus represent a new form of cell-cell communication important for driving tumor invasion that could be targeted to prevent metastasis.

cell biology↗

bioRxiv2024

Specialized signaling centers direct cell fate and spatial organization in a limb organoid model

Specialized signaling centers orchestrate robust development and regeneration. Limb morphogenesis, for instance, requires interactions between the mesoderm and the signaling center apical-ectodermal ridge (AER), whose properties and role in cell fate decisions have remained challenging to dissect. To tackle this, we developed mouse embryonic stem cells (mESCs)-based heterogeneous cultures and a limb organoid model, termed budoids, comprising cells with AER, surface ectoderm, and mesoderm properties. mESCs were first induced into heterogeneous cultures that self-organized into domes in 2D. Aggregating these cultures resulted in formation of limb bud-like structures in 3D, exhibiting chondrogenesis-based symmetry breaking and elongation. Using our organoids and quantitative in situ expression profiling, we uncovered that AER-like cells support nearby limb mesoderm and fibroblast identities while enhancing tissue polarization that permits distant cartilage formation. Together, our findings provide a powerful model to study aspects of limb morphogenesis, and reveal the ability of signaling center AER cells to concurrently modulate cell fate and spatial organization.

developmental biology↗

bioRxiv2024

Unveiling the dominant role of soil pH in shaping nitrogen cycling microbial gene abundances: Insights from 65-years of chemical fertilizer selection in acidic grassland meadow

Understanding the microbial processes driving the nitrogen (N) cycle is crucial for enhancing plant productivity and mitigating environmental pollution. The long-term application of synthetic fertilizers induces significant alterations in the microbial community functions. Microbes inhabiting acidic soils may exhibit distinct responses to chemical fertilizer application compared to soils with neutral pH. However, the chronic or occasional changes resulting from repeated nutrient flushes due to fertilizer application remain insufficiently elucidated, especially in acidic grasslands. Therefore, our study was on an acidic semi-natural grassland, where the soil was subjected to chemical fertilizer P (superphosphate), K (potassium sulfate), PK, N (ammonium nitrate), NPK, PK+N (PK applied in spring and N applied once in summer) over a span of 65 years. Gene abundances associated with the N-cycle (amoA, nifH, nirK, nirS, and nosZ) were quantified at seven different time points throughout the year considering the temporal effect caused by fertilizer application. Our findings reveal that in the long term, soil pH emerged as the predominant factor influencing the gene abundance related to N-fixation and denitrification outweighing the impact of nutrient availability. Notably, the application of N fertilizer had a positive effect on the abundance of nitrifiers, while the abundance of denitrifiers decreased due to soil acidification induced by fertilizer application. In summary, our study highlights that the microbial community involved in N cycling is more sensitive to the difference in soil pH shaped by long-term fertilizer application rather than to the direct impact of fertilizer application. HighlightO_LISeasonal sampling confirmed the long-term effect of fertilizer on microbial N-cycling genes C_LIO_LIFertilizer types shaped soil pH, consequently impacted on diazotrophs and denitrifies abundances C_LIO_LIAbundance of denitrifying microbe did not explain the increase of N2O emission by N fertilizer C_LIO_LIN2O emission was positively correlated with AOA abundance C_LI

microbiology↗

bioRxiv2024

Liming enhances the abundance and stability of nitrogen-cycling microbes: The buffering effect of long-term lime application

Lime application (liming) has historically been used to ameliorate soil acidity in grasslands. Liming effectively improves soil pH, plant productivity, and soil physicochemical properties, but the long-term impact of acidity control by liming on key microbial nitrogen (N)-cycling genes in semi-natural grasslands is unknown. We investigated the effect of 65 years of liming on N-cycling processes in the limed and control plots of the Ossekampen long-term grassland experiment in the Netherlands. These plots have not received any other fertilizers for 65 years. Soil sampling and nitrous oxide (N2O) emission measurements were conducted three times in spring and four times in summer, and quantitative real-time PCR was performed to determine the abundances of N-cycling genes, including ammonia-oxidation (amoA), denitrification (nirS, nirK, nosZ), and N-fixation (nifH) genes. Long-term liming increased the abundances of nitrifiers and denitrifiers but did not increase N2O emissions. Additionally, liming had a buffering effect that stabilized the population of N-cycling microbes against seasonal variations in abundance. Our results indicate that improving soil acidity through liming facilitates microbial N-cycling processes without increasing N2O emissions. HighlightsO_LI65 years of liming increased bacterial but not archaeal and fungal community abundance. C_LIO_LILiming increased the abundance of microbial N-cycling genes. C_LIO_LIThe buffering effect of liming reduced seasonal variations in the abundance of N microbes. C_LI

microbiology↗

bioRxiv2023

Direct visualization of emergent metastatic features within an ex vivo model of the tumor microenvironment

Metabolic conditions such as hypoxia, nutrient starvation, and media acidification, together with interactions with stromal cells are critical drivers of metastasis. Since these conditions arise deep within tumor tissues with poor access to the bloodstream, the observation of nascent metastases in vivo is exceedingly challenging. On the other hand, conventional cell culture studies cannot capture the complex nature of metastatic processes. We thus designed and implemented an ex vivo model of the tumor microenvironment to study the emergence of metastatic features in tumor cells in their native 3-dimensional (3D) context. In this system, named 3MIC, tumor cells spontaneously create ischemic-like conditions, and it allows the direct visualization of tumor-stroma interactions with high spatial and temporal resolution. We studied how 3D tumor spheroids evolve in the 3MIC when cultured under different metabolic environments and in the presence or absence of stromal cells. Consistent with previous experimental and clinical data, we show that ischemic environments increase cell migration and invasion. Importantly, the 3MIC allowed us to directly observe the emergence of these pro-metastatic features with single-cell resolution allowing us to track how changes in tumor motility were modulated by macrophages and endothelial cells. With these tools, we determined that the acidification of the extracellular media was more important than hypoxia in the induction of pro-metastatic tumor features. We also illustrate how the 3MIC can be used to test the effects of anti-metastatic drugs on cells experiencing different metabolic conditions. Overall, the 3MIC allows us to directly observe the emergence of metastatic tumor features in a physiologically relevant model of the tumor microenvironment. This simple and cost-effective system can dissect the complexity of the tumor microenvironment to test perturbations that may prevent tumors from becoming metastatic.

cancer biology↗

bioRxiv2022

Hypoxia potentiates the inflammatory fibroblast phenotype promoted by pancreatic cancer cell-derived cytokines

Cancer-associated fibroblasts (CAFs) are a major cell type in the stroma of solid tumors and can exert both tumor-promoting and tumor-restraining functions. This functional heterogeneity is correlated with the existence of transcriptionally distinct subpopulations of CAFs. CAF heterogeneity is observed in pancreatic ductal adenocarcinoma (PDAC), a tumor characterized by a remarkably dense and hypoxic stroma that features tumor-restraining myofibroblastic CAFs (myCAFs) and tumor-supporting inflammatory CAFs (iCAFs). While CAF heterogeneity can be driven in part by tumor cell-produced cytokines, other determinants shaping CAF identity and function are largely unknown. In vivo, we found that iCAFs display a hypoxic gene expression and biochemical profile and are enriched in hypoxic regions of PDAC tumors. Hypoxia leads fibroblasts to acquire an inflammatory gene expression signature and synergizes with cancer cell-derived cytokines to promote an iCAF phenotype in a HIF-1 dependent fashion. Furthermore, we show that HIF-1 stabilization is sufficient to induce an iCAF phenotype in stromal cells introduced into PDAC organoid co-cultures and to promote PDAC tumor growth. These findings indicate hypoxia-induced HIF-1 as a regulator of CAF heterogeneity and promoter of tumor progression in PDAC.

cell biology↗

bioRxiv2022

Failure of digit tip regeneration in the absence of suggests Lmx1b functions disparate from dorsoventral polarity

Mammalian digit tip regeneration is linked to the presence of nail tissue, but a nail-explicit model is missing. Here, we report that nail-less double-ventral digits of {Delta}LARM1/2 mutants that lack limb-specific Lmx1b enhancers fail to regenerate. To separate the nails effect from the lack of DV polarity, we also interrogate double-dorsal double-nail digits and show that they regenerate. Thus, DV polarity is not a prerequisite for regeneration and the nail requirement is supported. Transcriptomic comparison between wild-type and non-regenerative{Delta} LARM1/2 mutant blastemas reveals differential up-regulation of vascularization and connective tissue functional signatures in wild-type versus upregulation of inflammation in the mutant. These results, together with the finding of uniform Lmx1b expression in the wild-type blastema and in the dorsal dermis underneath the nail, indicate that, in addition of the nails effect, a direct role for Lmx1b in driving the progression of digit tip regeneration is likely.

developmental biology↗

bioRxiv2020

Defective membrane repair machinery impairs survival of invasive cancer cells

Cancer cells are able to reach distant tissues by migration and invasion processes. Enhanced ability to cope with physical stresses leading to cell membrane damages may offer to cancer cells high survival rate during metastasis. Consequently, down-regulation of the membrane repair machinery may be a therapeutic avenue for inhibiting metastasis. We show that migration of MDA-MB-231 cells on collagen I fibrils induces disruptions of plasma membrane and pullout of membrane fragments in the wake of cells. These cells are able to reseal membrane damages thanks to annexins (Anx) that are highly expressed in invasive cancer cells. In vitro membrane repair assays reveal that MDA-MB-231 cells respond heterogeneously to membrane injury and some of them possess very efficient repair machinery. Finally, we show that silencing of AnxA5 and AnxA6 leads to major defect of the membrane repair machinery responsible for the death of migrating MDA-MB-231 cells. Inhibition of membrane repair machinery may therefore represent a promising avenue for annihilating cancer metastasis. SummaryCancer cells are able to reach distant tissues by migration and invasion processes. This study shows that inhibition of the plasma membrane repair machinery may represent a promising avenue for annihilating cancer metastasis.

cancer biology↗