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Roux, C.

Publications and source records attributed to Roux, C..

At least 19 recordsLinked to original sources

Cutting-edge microfluidic technology allows detection and sorting of single fluorescent extracellular vesicles

Extracellular vesicles (EV) are cell-secreted, nanosized biological particles that are involved in most physiological and pathophysiological processes. Their composition and properties reflect the pathophysiological state of the producing cell, tissue, and organ. Released into biological fluids, they may be used as a potential source of non-invasive biomarkers for diagnosis, prognosis or therapeutic follow-up in several diseases such as cancer. However, despite the development of single-EV analysis technologies, the sorting of EV subpopulations for downstream analysis remains a major challenge. Here, we describe a novel instrument that combines a highly sensitive optical detection system with an advanced lab-on-a-chip device. Using engineered fluorescent small extracellular vesicles (sEV), we demonstrate that the Hekat NanoSorter enables sensitive, scalable, and reproducible detection and sorting of individual sEVs for downstream analysis by cryo-EM. Furthermore, detailed characterization of the sorted EV populations, together with the detection of sEVs labeled with an in vitro pathophysiological marker, demonstrates the ability of the NanoSorter to monitor and enrich clinically relevant EV-associated biomarkers. These findings highlight the clinical potential of extracellular vesicle sorting for their use as biomarkers for therapeutic monitoring and treatment outcome assessment.

bioengineering↗

Parallel differentiation extends beyond chromosomal inversions to small genomic islands on a background of rampant gene flow in the European flat oyster (Ostrea edulis)

Heterogeneous genomic landscapes of differentiation can reveal evolutionary processes acting over different timescales. Whereas genetic structure in the European flat oyster, Ostrea edulis, is mainly shaped by geography, previous work suggested parallel differentiation between western and eastern Mediterranean populations and between Atlantic and North Sea populations, resulting in genetic convergence between range margin populations in some genomic regions. Here, we analyzed 27 oysters from four locations using moderate-coverage (~20X) short-read whole-genome sequencing. We first examined three known chromosomal inversions and found evidence that they predate divergence with the sister species O. chilensis. We then scanned the genome for parallel differentiation at the individual-SNP rather than sliding-window scale using the product of allele-frequency differences across the two geographic contrasts. Two of the three inversions were detected, but we also identified 12 small non-inverted genomic regions showing even stronger signals of parallel differentiation. These 'islands of parallelism' typically overlap one or a few genes, and most showed elevated dXY consistent with long-term barriers to gene flow. Two islands overlap nuclear ATP synthase subunits, while mitochondrial ATP8 shows a convergent amino-acid change, suggesting possible nuclear-nuclear and mitonuclear incompatibilities. Finally, demographic inference with heterogeneous gene flow supported post-glacial secondary contact and a lineage-based rather than geography-based population topology. Our results suggest that the genomes of O. edulis lineages have been largely porous to gene flow upon secondary contact, except for chromosomal inversions and small genomic islands. These regions likely act as barrier loci, preserving a signal of a shared evolutionary history among marginal populations that has otherwise been erased by rampant gene flow.

evolutionary biology↗

Experimental evolution at high temperature in a Lepidopteran host of an entomopathogenic nematode and its associated microbiota

Entomopathogenic Nematodes (EPNs) have a great potential to control various insect crop pests, but their ability to perform their entire life-cycle in insects depends on environmental conditions. The insect killing ability of the endosymbiotic bacterial species (i.e., Xenorhabdus) located in the EPNs' gut (Steinernema) has been described for years. More recently, experimental data have suggested that other bacteria species of the nematode microbiota may also contribute to the parasitic success of the EPNs. EPNs and their associated bacteria should therefore be considered as holobionts. Global change, among its many predicted consequences, is expected to increase temperatures which may impair the parasitic success of these EPNs in the insect. Furthermore, higher environmental temperatures could modify the composition of the EPN-associated microbiota and could therefore also contribute to parasitic impairement. Using a S. carpocapsae strain originating from a French orchard, we investigated these hypotheses by experimentally evolving independent EPNs lineages in vivo in the lepidopteran species Galleria mellonella, at elevated and control temperatures. Several life-history traits of the evolved lineages were measured (insect killing ability, EPN emergence rate, EPN reproduction rate), and the bacterial microbiota of the evolved lineages was characterized by a metabarcoding approach. The EPNs' lineages evolved at higher temperature showed impaired emergence rate and their reproduction rate was lower, when compared to lineages evolved at control temperature. The bacterial microbiota composition of these lineages was modified, with particular enrichment in bacteria of the genera Alcaligenes and Pseudomonas. Our results provide new evidence that modification of the environment (here, the abiotic parameter temperature) during several successive generations can modify the parasitic success of EPNs, concomitantly to a change in the microbiota composition. A better understanding of the life-history traits of these EPNs should help improve the biocontrol efficiency in a context of global warming.

microbiology↗

Consequences of intra-locus recombination for branch-length-based inference of gene flow

Phylogenomic methods provide a powerful way to study introgression across broad clades of the tree of life, because they can test for gene flow from gene trees without requiring population-level resequencing data. These methods generally assume that each locus can be represented by a single non-recombining genealogy, which may be violated when recombination occurs within loci. Here, we used coalescent simulations to evaluate how intra-locus recombination affects gene-flow inferences in Aphid, a method using branch lengths to distinguish gene flow from incomplete lineage sorting in species triplets. Across the conditions tested, Aphid accurately recovered the proportion of loci affected by recent and intermediate gene flow, while recombination reduced the underestimation observed when gene flow is ancient. It also retained a relative timing signal, with accuracy decreasing as gene flow became older. This relative-timing approach was then applied to 456 African cichlid exon trees, where proposed gene flow involving Coptodon was consistently associated with intermediate-to-old rather than recent gene flow. Overall, our simulations suggest that intra-locus recombination does not increase error in Aphids inference of the prevalence of gene flow under the conditions tested, but can reduce temporal resolution for intermediate and ancestral events. When applied to cichlids, we show that this loss of resolution still permits the distinction between recent and older gene-flow.

evolutionary biology↗

Genomics of speciation in a great speciator (Aves: Zosterops) reveals the roles of both natural and sexual selection

Examining patterns of differentiation among lineage pairs at different stages of the speciation continuum can help disentangle the effects of linked and divergent selection, and identify lineage-specific targets of selection that may act as barrier loci during speciation. Here, we developed this approach using genomic data from a set of African and Indian Ocean belonging to the Zosteropidae family, commonly referred to as a "Great Speciator". We sampled a divergence continuum aiming to identify candidate barrier loci in phenotypically, genetically distinct, and parapatrically distributed, forms from a species complex, through comparisons with closely related species. Using three independent approaches, we showed that divergence at selected loci between geographic forms is mainly concentrated on the Z chromosome, unless forms differ in their ecologies. Functional annotation revealed that genes linked to song formation and immunity could be involved in reproductive isolation between forms only separated by physical barriers, while it is mostly genes related to adaptation to local conditions, morphology, and song behaviour that may explain reproductive isolation between forms with different ecologies, highlighting a role for combined effects of natural and sexual selection in driving the evolution of reproductive barriers in this rapidly speciating lineage

genomics↗

KDM5-driven transcriptional noise fuels plasticity-led awakening and relapse in paediatric cancer

How drug-tolerant persister (DTP) cells escape quiescence to drive tumour relapse is a central unresolved question in cancer evolution. Here, we identify transcriptional noise (TN), defined as the stochastic variability in gene expression, as a latent property of paediatric cancer cells that becomes a driver of adaptive regrowth after treatment withdrawal. Using functional assays, lineage tracing, single-cell transcriptomics, and multiscale landscape modelling, we show that therapy enriches mesenchymal-like tolerant states in neuroblastoma without clonal selection, while post-treatment awakening is a stochastic process fuelled by noise-enabled plasticity in cell-identity programmes. The histone demethylase KDM5A relocates to noisy cell-state genes during awakening, promoting H3K4me3 removal and chromatin remodelling at these loci. KDM5 inhibition abrogates this process, and suppresses transcriptional noise, halts DTP exit, and prevents tumour recovery in both neuroblastoma and hepatoblastoma models. These results establish DTP as an exploitable evolutionary bottleneck, positioning KDM5-mediated transcriptional noise as an actionable therapeutic target to limit cancer adaptation and relapse GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=120 SRC="FIGDIR/small/682004v1_ufig1.gif" ALT="Figure 1"> View larger version (34K): org.highwire.dtl.DTLVardef@edba70org.highwire.dtl.DTLVardef@16b1bd9org.highwire.dtl.DTLVardef@f153acorg.highwire.dtl.DTLVardef@1ac5a4e_HPS_FORMAT_FIGEXP M_FIG C_FIG

cancer biology↗

Soil phosphate availability modulates the arbuscular mycorrhizal fungal community and mycorrhizal nutrition in wheat

O_LIUnderstanding the contributions of arbuscular mycorrhizal fungi (AMF) to plant nutrition is essential for sustainable agriculture. We hypothesised that soil phosphorus (P) availability modulates the diversity and functionality of wheat root-associated AMF community, particularly the mycorrhizal nutrition. C_LIO_LIWheat plants were sampled over two campaigns (2019 and 2022) in a long-term P fertilisation trial. The expression of the wheat transporters involved in the mycorrhizal nutrition was assessed by RT-qPCR, and AMF community composition by metabarcoding. Complementary experiments under controlled conditions were performed to further study the interaction between nitrogen (N) and P availability on AM. C_LIO_LIDifferent effects of P on AMF colonisation and transporter expression were observed in field-grown wheat depending on the campaigns, which differed in wheat N status. Controlled experiments confirmed that AMF colonisation depends on the limitation of either P or N, but that regulation of peri-arbuscular phosphate, ammonium and nitrate transporters depends on the nature of the limiting soil nutrients. Additionally, AMF communities varied according to the soil P availability, with the Funneliformis genus becoming more dominant under high P conditions for both years. C_LIO_LITogether, our findings show that N and P availability jointly shape root AMF communities and AM functioning. Combining community profiling and molecular markers of colonisation and nutrition offers a framework to better understand AMF contributions to plant nutrition across agroecosystems. C_LI

plant biology↗

Untangling the contribution of adaptive versus non-adaptive processes in the evolution of reproductive isolation between Coenonympha butterflies.

Speciation is a key evolutionary process which has been studied in numerous organisms and at multiple scales, from lineage radiation to gene expression. However, the factors explaining the rise of new species are not yet fully understood, and the relative contribution of neutral versus selective evolutionary processes in triggering and maintaining reproductive isolation between lineages is still debated. To explore this question, we study the divergence of two butterfly species, Coenonympha arcania and C. gardetta (Nymphalidae), which diverged relatively recently but show strong ecological differences. Whole genome sequence data reveal high overall differentiation between the two lineages, best explained by a long period of isolation at the early stage of their divergence. Demographically explicit approaches identify that 6.6% of the genome (32.7 Mbp) is impermeable to gene flow between the two species. Lots of these barrier loci are located on the Z chromosome, potentially spanning 75% of its length which would indicate that a large Z effect is at play in this speciation. Moreover, only a small proportion of barriers showed signatures of selection, suggesting that non-adaptive processes largely contributed to the build-up of reproductive isolation. Therefore, although genes involved in stress response and response to hypoxia are interesting candidates under selection, the adaptation of C. gardetta to alpine conditions may not to be the main driver of speciation. Our study brings an original example of intertwined adaptive and non-adaptive processes leading to reproductive isolation in a speciation with secondary contact and improves our understanding of the genomic underpinnings of species divergence.

evolutionary biology↗

Constitutive expression of Cas9 and rapamycin-inducible Cre recombinase facilitates conditional genome editing in Plasmodium berghei

Malaria is caused by protozoan parasites of the genus Plasmodium and remains a global health concern. The parasite has a highly adaptable life cycle comprising successive rounds of asexual replication in a vertebrate host and sexual maturation in the mosquito vector Anopheles. Genetic manipulation of the parasite has been instrumental for deciphering the function of Plasmodium genes. Conventional reverse genetic tools cannot be used to study essential genes of the asexual blood stages, thereby necessitating the development of conditional strategies. Among various such strategies, the rapamycin-inducible dimerisable Cre (DiCre) recombinase system emerged as a powerful approach for conditional editing of essential genes in human-infecting P. falciparum and in the rodent malaria model parasite P. berghei. We previously generated a DiCre-expressing P. berghei line and validated it by conditionally deleting several essential asexual stage genes, revealing their important role also in sporozoites. The advent of CRISPR enabled targeted genome editing with higher accuracy and specificity and greatly advanced genome engineering in Plasmodium spp. Here, we developed new P. berghei parasite lines by integrating the DiCre cassette and a fluorescent marker in parasites constitutively expressing Cas9. Owing to the dual integration of CRISPR-Cas9 and DiCre, these new lines allow unparalleled levels of gene modification and conditional regulation simultaneously. To illustrate the versatility of this new tool, we conditionally knocked-out the essential gene encoding the claudin-like apicomplexan micronemal protein (CLAMP) in P. berghei and confirm the role of CLAMP during invasion of erythrocytes.

microbiology↗

Diverging Arabidopsis populations quickly accumulate pollen-acting genetic incompatibilities

The process by which species diverge from one another, gradually accumulate genetic incompatibilities and eventually reach full-fledged reproductive isolation is a key question in evolutionary biology. However, the nature of reproductive barriers, the pace at which they accumulate and their genomic distribution remain poorly documented. The disruption of co-adapted epistatic interactions in hybrids and the accumulation of selfish genetic elements are proposed contributors to this process, and can lead to the distortion of the mendelian segregation of the affected loci across the genome. In this study we detect and quantify segregation distortion across the genomes of crosses produced from a diverse sampling of Arabidopsis lyrata and A. halleri populations, two species at the early stages of speciation and that can still interbreed. We show that both the frequency of occurrence and the magnitude of distortion loci increase as the parents genetic distance from one another increases. We also observe that distorter loci evolve rapidly, as they occur not only within interspecific hybrids, but also in intraspecific hybrids produced from isolated population crosses. Finally, we identify both genome-wide non-independence and two specific genomic regions on different chromosomes where opposite distortion effects are repeatedly observed across multiple F1 individuals, suggesting negative epistasis is a major contributor to the evolution of hybrid segregation distortion. Our study demonstrates that pollen-acting segregation distortion is ubiquitous, and contributes not only to the ongoing reproductive isolation between A. halleri and A. lyrata, but also between very recently diverged populations of the same species.

genomics↗

Social evolution in termites reduces natural selection efficacy

In eusocial species, reproduction is monopolized by a few reproductive individuals. From the perspective of population genetics, this implies that the effective population size (Ne) of these organisms is likely to be smaller compared to solitary species, as has been proposed in the literature for eusocial hymenoptera. In this study, we investigate the genomic consequences of eusociality in termites (Isoptera) on two different timescales. First, by analyzing transcriptome data from 66 Blattodea species, we focus on the ratio of non-synonymous to synonymous mutations (dN/dS) as a marker of natural selection efficiency and effective population size. Our results demonstrate an elevated dN/dS ratio in termites compared to other members of Blattodea, further generalizing the idea that convergent evolution toward eusociality strongly reduces the effective population size and the genome-wide efficiency of natural selection. Then, by comparing 68 termite transcriptomes, we show that this decrease in natural selection efficiency is even more pronounced in termites displaying high levels of social complexity. This study contributes to understanding the complex interplay between social structures and natural selection patterns, highlighting the genetic footprint of eusociality in shaping the evolution of termites.

evolutionary biology↗

Dynamic Plasticity Systems Direct Early Adaptation to Treatment in Neuroblastoma

Neuroblastoma, like many aggressive cancers, exhibits phenotypic heterogeneity, contributing to therapy resistance and disease progression. However, direct evidence of how phenotypic plasticity influences tumour evolution remains limited. Using a multi-resolution quantitative approach, we define the principles, types, and dynamics of plasticity in neuroblastoma at unprecedented resolution. We demonstrate that intrinsic plasticity is a model-dependent process that enables the coexistence of drug-sensitive and drug-resistant states in environmentally stable conditions, positioning plasticity as a bet-hedging strategy that allows tumours to anticipate environmental changes. Additionally, we show plasticity varies across lineages and single-cell-derived clones, establishing that it is not merely an induced response but a heritable and selectable trait. By simultaneously mapping plasticity at and clonal dynamics in evolving neuroblastoma populations, we show plasticity is shaped by selective pressures, reinforcing its role as a fundamental driver of neuroblastoma evolution in a treatment- and genetic background-dependent manner. We define three distinct modes of plasticityled adaptation. In the selection of the plasticity model, strong selective pressure temporarily constrains phenotypic transitions, but they reemerge with greater dynamics once the stressor is removed, favouring the selection of highly plastic clones. In the adaptive plasticity model, phenotypic transitions actively reshape tumour heterogeneity, allowing for rapid adaptation to treatment while minimising the impact of clonal selection. Finally, in the plasticity equilibrium model, phenotypic transitions persist at baseline, maintaining a state of phenotypic fluidity, with clonal selection ultimately dictating tumour evolution. These findings highlight the diverse, context-dependent strategies that neuroblastoma populations employ to navigate selective pressures and therapy resistance, emphasizing the need for plasticity-targeting therapeutic approaches to disrupt tumour adaptation and improve treatment outcomes. TeaserNatural selection and phenotypic transitions shape adaptive evolution, guiding neuroblastoma survival under treatment pressure.

cancer biology↗

Parallel on-chip micropipettes enabling quantitative multiplexed characterization of vesicle mechanics and cell aggregates rheology

Micropipette aspiration (MPA) is one of the gold standards to quantify biological samples mechanical properties, which are crucial from the cell membrane scale to the multicellular tissue. However, relying on the manipulation of individual home-made glass pipettes, MPA suffers from low throughput and difficult automation. Here, we introduce the sliding insert micropipette aspiration (SIMPA) method, that permits parallelization and automation, thanks to the insertion of tubular pipettes, obtained by photolithography, within microfluidic channels. We show its application both at the lipid bilayer level, by probing vesicles to measure membrane bending and stretching moduli, and at the tissue level by quantifying the viscoelasticity of 3D cell aggregates. This approach opens the way to high-throughput, quantitative mechanical testing of many types of biological samples, from vesicles and individual cells to cell aggregates and explants, under dynamic physico-chemical stimuli.

biophysics↗

Rapid establishment of species barriers in plants compared to animals

Speciation, the process through which new reproductively isolated species emerge from ancestral populations, occurs due to the gradual accumulation of barriers to gene flow within genomes. To date, the notion that interspecific genetic exchange occurs more frequently between plant species than animals species has gained a strong footing in the scientific discourse. By examining the dynamics of gene flow across a continuum of divergence in both kingdoms, we observe the opposite relationship: plants experience less introgression than animals at the same level of genetic divergence, suggesting that species barriers are established more rapidly in plants. This pattern raises questions about which differences in microevolutionary processes between plants and animals influence the dynamics of reproductive isolation establishment at the macroevolutionary scale. One Sentence SummaryGenetic exchange is more frequent between animal species than plants, challenging historical views.

evolutionary biology↗

Climate change promoted allopatric divergence and explains the current disjunct geographic distribution of the Ophrys aveyronensis species complex (Orchidaceae)

AimSpecies with disjunct geographic distributions provide natural opportunities to investigate incipient or recent allopatric divergence. The combination of both genetic and ecological data may be fruitful to decipher the causes of such pattern: i) actual vicariance, ii) successful colonization from one source range to a new range (dispersal, biological introduction) or iii) parallel convergent evolution. LocationSouthern France and Northern Spain. TaxonThe bee orchid Ophrys aveyronensis (and its two recognized subspecies O. a. subsp. aveyronensis and O. a. subsp. vitorica) displays a disjunct geographic distribution with two subranges separated by 600 km on both sides of the Pyrenees Mountains. MethodsThrough a combination of population genomics and Ecological Niche Modelling (ENM), we investigated the causes of this intriguing biogeographic pattern. ResultsThe population genomic data demonstrate that all the studied populations exhibit similar patterns of genetic diversity and dramatic decrease in effective size compared to the ancestral population. Significant genetic differentiation and reciprocal monophyly exist between populations of the two subranges of O. aveyronensis, despite a very recent divergence time as young as ca. 1500 generations ago. Moreover, (paleo-)ecological Niche Modelling analyses support that the disjunct geographic distribution of the O. aveyronensis is consistent with a range split of a broad ancestral range, contraction and distinct longitudinal and latitudinal shifts in response to climate warming during the Holocene. Main ConclusionThe congruence of the results obtained from both population genomics and ENM approaches documents how very recent continental allopatric divergence initiated speciation in this system. Ophrys aveyronensis provides a promising opportunity to study the onset of reproductive isolation and parallel evolution following an initial stage of geographic separation in a group with high diversification rate.

evolutionary biology↗

The effects of environmental heterogeneity within cities on the evolution of clines

O_LIThere is increasing evidence that environmental change associated with urbanization can drive rapid adaptation. However, most studies of urban adaptation have focused on coarse urban vs. rural comparisons or sampled along a single urban-rural environmental gradient, thereby ignoring the role that within-city environmental heterogeneity might play in adaptation to urban environments. C_LIO_LIIn this study, we examined fine-scale variation in the presence of HCN--a potent anti-herbivore defense--and its two underlying genes (Ac and Li) between park green spaces and surrounding suburban habitats for five city parks in the Greater Toronto Area. C_LIO_LIWe show that fine-scale urbanization has driven the formation of micro-clines in HCN on a scale of < 2 km, though the presence and strength of micro-clines varied across parks. Interestingly, these micro-clines were driven by lower HCN frequencies inside park green spaces, and are therefore in the opposite direction to that predicted based on previously described patterns of HCN frequency change along urban-rural gradients. C_LIO_LISynthesis: These results suggest larger scale, adaptive urban-rural clines occur across a complex matrix of environmental heterogeneity within cities that drives fine-scale adaptive microclines of varying strengths and directions. C_LI

evolutionary biology↗

Introgression between highly divergent sea squirt genomes: an adaptive breakthrough?

AO_SCPLOWBSTRACTC_SCPLOWHuman-mediated introductions are reshuffling species distribution on a global scale. Consequently, an increasing number of allopatric taxa are now brought into contact, promoting introgressive hybridization between incompletely isolated species and new adaptive gene transfer. The broadcast spawning marine species, Ciona robusta, has been recently introduced in the native range of its sister taxa, Ciona intestinalis, in the English Channel and North-East Atlantic. These sea squirts are highly divergent, yet hybridization has been reported by crossing experiments and genetic studies in the wild. Here, we examined the consequences of secondary contact between C. intestinalis and C. robusta in the English Channel. We produced genomes phased by transmission to infer the history of divergence and gene flow, and analyzed introgressed genomic tracts. Demographic inference revealed a history of secondary contact with a low overall rate of introgression. Introgressed tracts were short, segregating at low frequency, and scattered throughout the genome, suggesting traces of past contacts during the last 30 ky. However, we also uncovered a hotspot of introgression on chromosome 5, characterized by several hundred kb-long C. robusta haplotypes segregating in C. intestinalis, that introgressed during contemporary times the last 75 years. Although locally more frequent than the baseline level of introgression, C. robusta alleles are not fixed, even in the core region of the introgression hotspot. Still, linkage-disequilibrium patterns and haplotype-based tests suggest this genomic region is under recent positive selection. We further detected in the hotspot an over-representation of candidate SNPs lying on a cytochrome P450 gene with a high copy number of tandem repeats in the introgressed alleles. Cytochromes P450 are a superfamily of enzymes involved in detoxifying exogenous compounds, constituting a promising avenue for functional studies. These findings support that introgression of an adaptive allele is possible between very divergent genomes and that anthropogenic hybridization can provide the raw material for adaptation of native lineages in the Anthropocene.

evolutionary biology↗

Inferring the demographic history of tetraploid species from genomic data

Genomic patterns of diversity and divergence are impacted by certain life history traits, reproductive systems and demographic history. The latter is characterised by fluctuations in population sizes over time, as well as by temporal patterns of introgression. For a given organism, identifying a demographic history that deviates from the standard neutral model allows a better understanding of its evolution, but also helps to reduce the risk of false positives when screening for molecular targets of natural selection. Tetraploid organisms and beyond have demographic histories that are complicated by the mode of polyploidisation, the mode of inheritance and different scenarios of gene flow between subgenomes and diploid parental species. Here we provide guidelines for experimenters wishing to address these issues through a flexible statistical framework: approximate Bayesian computation (ABC). The emphasis is on the general philosophy of the approach to encourage future users to exploit the enormous flexibility of ABC beyond the limitations imposed by generalist data analysis pipelines.

genomics↗