Biology subjects

Conti, L.

Publications and source records attributed to Conti, L..

16 recordsLinked to original sources

bioRxiv2026

Functional validation of allele-specific LMNB1 silencing in patient-derived astrocytes as a therapeutic option for Autosomal Dominant Leukodystrophy

Adult-onset Autosomal Dominant Leukodystrophy (ADLD) is a rare fatal leukodystrophy caused by increased LMNB1 gene dosage, most commonly resulting from duplication of the LMNB1 locus. Because ADLD is a gene dosage disorder, selective reduction of pathological LMNB1 expression represents a rational therapeutic strategy. Although allele-specific RNA interference has previously been shown to lower LMNB1 levels in patient-derived fibroblasts and directly reprogrammed neurons, its therapeutic effects have not been evaluated in disease-relevant human glial cells or using functional efficacy endpoints. Here, we established human induced pluripotent stem cell-derived astrocytes from ADLD patients as a human glial model in which to validate allele-specific LMNB1 silencing across molecular, cellular, and functional readouts. ADLD astrocytes recapitulated increased LMNB1 expression and characteristic nuclear abnormalities and displayed transcriptional alterations affecting extracellular matrix organization, calcium homeostasis, metabolism and RNA processing. Functionally, these cells also exhibited functional phenotypes suitable for therapeutic evaluation: astrocyte-conditioned medium impaired the viability of both murine and human oligodendroglial cultures, while conditioned-medium and direct astrocyte-seeding paradigms revealed impaired post-lesion myelin recovery in lysolecithin-treated cerebellar organotypic slices. Allele-specific LMNB1 silencing restored physiological LMNB1 levels, corrected nuclear abnormalities, attenuated astrocyte-mediated oligodendroglial toxicity, improved post-lesion myelin recovery, and was associated with selective transcriptional programs associated with extracellular support and cholesterol metabolism. Together, these findings provide molecular, cellular, and functional validation of allele-specific LMNB1 dosage correction in patient-derived human astrocytes and offer key support for LMNB1-lowering strategies in disease-relevant human glial cells.

neuroscience↗

bioRxiv2026

ARHGEF6-dependent cytoskeletal regulation underlies a conserved program of forebrain interneuron development

The molecular programs coordinating inhibitory interneuron migration, maturation, and survival during forebrain development remain incompletely understood. Here we investigate ARHGEF6, a RAC1/CDC42 guanine nucleotide exchange factor linked to X-linked intellectual disability (XLID46) and previously studied only at postsynaptic compartments, and reveal an earlier, conserved role in forebrain interneuron development. ARHGEF6 is selectively enriched in the inhibitory lineage during the peak of interneuron generation and migration. Its loss in mice reduces the number of cortical and hippocampal interneurons, disrupts tangential migration, increases developmental cell death, and impairs morphological and electrophysiological maturation. Strikingly, ARHGEF6-knockout human iPSC-derived organoids and assembloids mirror these deficits exhibiting increased apoptosis, reduced neuronal output, disorganized growth cones, impaired neurite branching, and disrupted migratory dynamics. These cross-species findings reframe ARHGEF6 as an early, essential orchestrator of inhibitory circuit assembly and reveal a conserved cytoskeletal program whose disruption produces the excitatory-inhibitory imbalance linked to cognitive dysfunction.

developmental biology↗

bioRxiv2025

Human liver organoids uncover ductular reaction in Wolman disease and inform gene therapy design

Wolman disease (WD), the severe infantile form of lysosomal acid lipase deficiency, is a rare metabolic disorder caused by inactivating mutations in the LIPA gene. Although WD is characterized by profound hepatic dysfunction, experimental human systems capable of modelling multicellular liver pathology and supporting therapeutic testing remain limited. Here, we generated an isogenic human model of WD by introducing LIPA loss-of-function mutations into induced pluripotent stem cells and differentiating them into multicellular human liver organoids (HLO). LIPA-deficient HLO preserved hepatic lineage specification while recapitulating key biochemical and cellular features of WD, including loss of LIPA activity, lysosomal expansion, lipid accumulation, and activation of inflammatory and fibrogenic programs. Single-cell RNA sequencing resolved cell-type-specific disease states across hepatocyte-, stromal-, and biliary-like populations, revealing the emergence of a reactive biliary program consistent with ductular reaction, a complex tissue response associated with chronic liver injury. Importantly, this reactive biliary phenotype was supported by targeted gene-expression analysis in WD liver organoids and independently validated in liver tissue from mouse models and WD patients. Isolated LIPA-deficient cholangiocyte organoids failed to reproduce the DR-associated program, indicating that this response depends on multicellular interactions within the hepatic microenvironment rather than on biliary cell-autonomous dysfunction alone. Consistently, hepatocyte-directed AAV-mediated restoration of LIPA expression attenuated metabolic stress, inflammatory and fibrogenic programs, and suppressed ductular reaction both in organoids and in vivo. Together, these findings establish multicellular human liver organoids as a physiologically relevant platform for modelling emergent tissue-level responses in WD and for evaluating therapeutic rescue strategies in a human context.

pathology↗

bioRxiv2024

Abscisic acid and GIGANTEA signalling converge to regulate the recruitment of CONSTANS to the FT promoter and activate floral transition

Plants align flowering with optimal seasonal conditions to increase reproductive success. This process depends on modulating signalling pathways that respond to diverse environmental and hormonal inputs, thereby regulating the transition to flowering at the shoot apical meristem. In Arabidopsis, long-day photoperiods (LDs) stimulate the transcription of FLOWERING LOCUS T (FT), encoding the main florigenic signal. FT activation is mediated by the transcriptional regulator CONSTANS (CO), which binds to the CO responsive elements (COREs) located in the proximal FT promoter region. The phytohormone abscisic acid also (ABA) contributes to FT activation together with GIGANTEA (GI) to regulate drought escape (DE). Whether CO is a target of ABA and GI actions for the regulation of FT is, however, unknown. Here, we report that ABA and its signalling components promote CO recruitment to the CORE1/2, without causing reductions in the diel pattern of CO protein accumulation. ChIPseq analyses show that ABA broadly shapes the CO DNA binding landscape, which is enriched at the promoters of genes involved in the response to abiotic stress. We also found that GI promotes CO recruitment to the CORE1/2 region, and that CO recruitment is required for the accumulation of RNAPol II at the transcription start site of FT. Finally, we show that GI and ABA signalling pathways are largely epistatic in the control of flowering time, suggesting their involvement in the same molecular process. Taken together, these observations suggest that varying water deficit conditions modulate CO recruitment and FT expression, thus dictating DE strategies in Arabidopsis. HighlightABA and GIGANTEA signalling promote FLOWERING LOCUS T (FT) transcriptional activation by regulating the binding of the transcription factor CONSTANS to the proximal FT promoter.

plant biology↗

bioRxiv2024

Generation and benchmarking of a collection of hiPSC lines from Schizophrenia Patients with Diverse Clinical Profiles

Limited therapeutic advancements in Schizophrenia (SCZ) depend on the heterogeneous nature of the disorder, impacting drug development and clinical trials that assume uniform therapy response, neglecting individual genetic and epigenomic variability. Disease modeling using human induced pluripotent stem cells (hiPSCs) is ideally suited for precision medicine, enabling individualized treatment approaches. Here, we describe the generation of patient-specific lines from somatic cells of SCZ individuals with well-defined diverse clinical trajectories using a Sendai virus-based reprogramming system. Karyotypically and CGH-array validated, the generated hiPSCs expressed diagnostic markers and demonstrated functional pluripotency. Converting these hiPSCs into neural progenitor cells enables the identification of aberrant cellular phenotypes associated with specific pathologically relevant neural phenotypes. This collection of hiPSC lines serves as a platform for developing therapeutic compounds targeting neural populations, potentially addressing early-stage disease alterations.

neuroscience↗

bioRxiv2023

"Flower power": how flowering affects spectral diversity metrics and their relationship with plant diversity

Biodiversity monitoring is constrained by cost- and labour-intensive field sampling methods. Increasing evidence suggests that remotely sensed spectral diversity (SD) is linked to plant diversity, holding promise for monitoring applications. However, studies testing such a relationship reported conflicting findings, especially in challenging ecosystems such as grasslands, due to their high temporal dynamism and variety. It follows that a thorough investigation of the key factors, such as the metrics applied (i.e., continuous, categorical) and phenology (e.g., flowering), influencing such a relationship is necessary. Thus, this study aims to assess the applicability of SD for plant diversity monitoring at the local scale by testing six different SD metrics while considering the effect of the presence of flowering on the relationship and resampling the original data to assess how spatial resolution affects the results. Taxonomic diversity was calculated based on data collected in 159 plots with 1.5 m x1.5 m experimental mesic grassland communities. Spectral information was collected using a UAV-borne sensor measuring reflectance across six bands in the visible and near-infrared range at [~]2 cm spatial resolution. Our results show that, in the presence of flowering, the relationship is significant and positive only when SD is calculated using categorical metrics. Despite the observed significance, the variance explained by the models had very low values, with no evident differences when resampling spectral data to coarser pixel sizes. Such findings suggest that new insights into the possible confounding effects on the SD[~]plant diversity in grassland communities are needed to use SD for monitoring purposes.

ecology↗

bioRxiv2022

The relationship between spectral and plant diversity: disentangling the influence of metrics and habitat types

Biodiversity monitoring is crucial for ecosystem conservation, yet field data collection is limited by costs, time, and extent. Remote sensing represents a convenient approach providing frequent, near-real-time information over wide areas. According to the Spectral Variation Hypothesis (SVH), spectral diversity (SD) is an effective proxy of environmental heterogeneity, which ultimately relates to plant diversity. So far, studies testing the relationship between SD and biodiversity have reported contradictory findings, calling for a thorough investigation of the key factors (e.g., metrics applied, ecosystem type) and the conditions under which such a relationship holds true. This study investigates the applicability of the SVH for plant diversity monitoring at the landscape scale by comparing the performance of three different types of SD metrics. Species richness and functional diversity were calculated for more than 2000 cells forming a grid covering the Czech Republic. Within each cell, we quantified SD using a Landsat-8 "greenest pixel" composite by applying: i) the standard deviation of NDVI, ii) Raos Q entropy index, and iii) richness of "spectral communities". Habitat type (i.e., land cover) was included in the models describing the relationship between SD and ground biodiversity. Both species richness and functional diversity show positive and significant relationships with each SD metric tested. However, SD alone accounts for a small fraction of the deviance explained by the models. Furthermore, the strength of the relationship depends significantly on habitat type and is highest in natural transitional areas. Our results underline that, despite the stability in the significance of the link between SD and plant diversity at this scale, the applicability of SD for biodiversity monitoring is context-dependent and the factors mediating such a relationship must be carefully considered to avoid drawing misleading conclusions. HighlightsO_LIPlant species richness and functional diversity show significant and positive relationships with spectral diversity C_LIO_LISpectral diversity alone explains a small fraction of the total variability in ground biodiversity C_LIO_LISlight differences among the performances of the spectral diversity metrics tested C_LIO_LIThe relationship between spectral and plant diversity is context-dependent C_LI

ecology↗

bioRxiv2022

Compositional profiling of extracellular vesicles/lipoproteins mixtures by AFM nanomechanical imaging

The widely overlapping physicochemical properties of lipoproteins (LPs) and extracellular vesicles (EVs) represents one of the main obstacles for the isolation and characterization of these pervasive biogenic lipid nanoparticles. We herein present the application of an atomic force microscopy (AFM)-based quantitative morphometry assay to the rapid nanomechanical screening of mixed LPs and EVs samples. The method can determine the diameter and the mechanical stiffness of hundreds of individual nanometric objects within few hours. The obtained diameters are in quantitative accord with those measured via cryo-electron microscopy (cryo-EM); the assignment of a specific nanomechanical readout to each object enables the simultaneous discrimination of co-isolated EVs and LPs even if they have overlapping size distributions. EVs and all classes of LPs are shown to be characterized by specific combinations of diameter and stiffness, thus making it possible to estimate their relative abundance in EV/LP mixed samples in terms of stoichiometric ratio, surface area and volume. As a side finding, we show how the mechanical behaviour of specific LP classes is correlated to distinctive structural features revealed by cryo-EM. To the best of our knowledge, these results represent the first systematic single-particle mechanical investigation of lipoproteins. The described approach is label-free, single-step and relatively quick to perform. Importantly, it can be used to analyze samples which prove very challenging to assess with several established techniques due to ensemble-averaging, low sensibility to small particles, or both, thus providing a very useful tool for quickly assessing the purity of EV/LP isolates including plasma- and serum-derived preparations.

biophysics↗

bioRxiv2022

Functional traits trade-offs define plant population stability worldwide

O_LIEcological theory posits that temporal stability patterns in plant populations are associated with differences in species ecological strategies. However, empirical evidence is lacking about which traits, or trade-offs, underlie species stability, specially across different ecosystems. C_LIO_LITo address this, we compiled a global collection of long-term permanent vegetation records (>7000 plots from 78 datasets) from a wide range of habitats and combined this with existing trait databases. We tested whether the observed inter-annual variability in species abundance (coefficient of variation) was related to multiple individual traits and multivariate axes of trait variations (PCoA axes). C_LIO_LIWe found that species with greater leaf dry matter content and seed mass were consistently more stable over time (lower variability in species abundance) although other leaf traits played a significant role as well, albeit weaker. Using multivariate axes did not improve predictions by specific traits. C_LIO_LIOur results confirm existing theory, providing compelling empirical evidence on the importance of specific traits, which point at ecological trade-offs in different resource use and dispersal strategies, on the stability of plant populations worldwide. C_LI

ecology↗

bioRxiv2021

Defective linear and circular RNAs biogenesis in Huntington's disease: CAG repeat expansion hijacks neuronal splicing

Alternative splicing (AS) appears to be altered in Huntingtons disease (HD), but its significance for early, pre-symptomatic disease stages has not been inspected. Here, taking advantage of Htt CAG knock-in mouse in vitro and in vivo models, we demonstrate a strong correlation between Htt CAG repeat length and increased aberrant linear AS, specifically affecting neural progenitors and, in vivo, the striatum prior to overt behavioral phenotypes stages. Remarkably, expanded Htt CAG repeats reflect on a previously neglected, global impairment of back-splicing, leading to decreased circular RNAs production in neural progenitors. Though the mechanisms of this dysregulation remain uncertain, our study unveils network of transcriptionally altered micro-RNAs and RNA-binding proteins (CELF, hnRNPS, PTBP, SRSF) which, in turn, might influence the AS machinery, primarily in neural cells. We suggest that this unbalanced expression of linear and circular RNAs might result in altered neural fitness, contributing to HD striatal vulnerability.

genetics↗

bioRxiv2021

LOTVS: a global collection of permanent vegetation plots

Analysing temporal patterns in plant communities is extremely important to quantify the extent and the consequences of ecological changes, especially considering the current biodiversity crisis. Long-term data collected through the regular sampling of permanent plots represent the most accurate resource to study ecological succession, analyse the stability of a community over time and understand the mechanisms driving vegetation change. We hereby present the LOng-Term Vegetation Sampling (LOTVS) initiative, a global collection of vegetation time-series derived from the regular monitoring of vascular plants in permanent plots. With 79 datasets from five continents and 7789 vegetation time-series monitored for at least six years and mostly on an annual basis, LOTVS possibly represents the largest collection of temporally fine-grained vegetation time-series derived from permanent plots and made accessible to the research community. As such, it has an outstanding potential to support innovative research in the fields of vegetation science, plant ecology and temporal ecology.

ecology↗

bioRxiv2021

Sticking around: plant persistence strategies on edaphic islands

O_LISpecies extinction risk at local scales can be partially offset by strategies promoting in-situ persistence. We explored how persistence-related traits of clonal and non-clonal plants in temperate dry grasslands respond intra- and interspecifically to variation in environmental conditions (soil, climate) and insularity. C_LIO_LIWe focused on edaphic island specialist species, hypothesizing that plants experiencing harsh soil environments and strong insularity are distinguished by traits supporting enhanced persistence, such as small stature, long lifespan and resource-conservative strategies. We used linear mixed-effect models and bivariate ordinary least squares linear models to explore the response of species triats to environmental and biogeographic predictors. C_LIO_LIWe found general support for this hypothesis. Soil properties and insularity emerged as the most important drivers of trait patterns. However, clonal species showed more consistent responses to variation in environmental conditions and insularity than non-clonal plants, which were characterized by distinct species-specific responses. C_LIO_LISoil properties and insularity confirmed their major role in shaping the persistence strategies of edaphic island plant species. These drivers may exert their effect on specific functions (e.g. belowground resource conservation captured by BDMC). Additionally, we unambiguously identified that clonal species had different persistence strategies than non-clonal ones. C_LI

ecology↗

bioRxiv2021

Insularity forcing on plant persistence strategies in edaphic island systems

AimTrait-based approaches are increasingly implemented in island biogeography, providing key insights into the eco-evolutionary dynamics of insular systems. However, what determines persistence of plant species once they have arrived and established in an island remains largely unexplored. Here, we examined links between non-acquisitive persistence strategies and insularity across three terrestrial edaphic island systems, hypothesising that insularity promotes strategies for local persistence. LocationEurope: Western Carpathians, Moravia, and Cantabrian Range. Time periodPresent. Major taxa studiedVascular plants. MethodsFor each system, we used linear models at the island scale to test whether persistence-related plant trait patterns (average trait values and diversity) depend on three insularity metrics (island size, isolation and target effect). We focused on patterns of edaphic island specialists because, in contrast to matrix-derived species, their presence is confined to the edaphic islands. ResultsWe found that insularity metrics explained large proportions in the variation of the average and diversity of persistence-related traits of edaphic island specialists. Insularity was associated with a decline in the proportion of island specialists that have clonal abilities, yet it affected trait values of specialists towards enhanced abilities to persist locally (e.g. more extensive lateral spread) while reducing trait variability. Higher degrees of insularity within the systems were translated to stronger effects on functional trait patterns. Main conclusionsInsularity affects plant species diversity, distribution and forms in terrestrial island-like systems, similarly as it is assumed for true islands. Insularity - measured using a single (island size, isolation) or combined (target effect) predictors - may operate selecting for enhanced and less diverse persistence strategies. Ultimately, this process, which we call insularity forcing, operates as a selective process to promote species ability to avoid local extinction and to persist on terrestrial islands.

ecology↗

bioRxiv2021

Climate fluctuations drive the recruitment and growth of temperate grassland plants

Recent climate warming is associated with the increasing magnitude and frequency of extreme events, including heatwaves and drought periods worldwide. Such events can have major effects on the species composition of plant communities, hence on biodiversity and ecosystem functioning. Here we studied responses of Central European dry grassland plants to fluctuating temperature and precipitation over the last thirty years with monthly temporal resolution. We assessed the seasonal and annual dynamics of plant recruitment and growth based on the analysis of annual growth rings from the root collar. Although most studies so far applied such methods to trees and shrubs, we focused on typical grassland plants, two forbs and two chamaephytes. We related the recruitment and annual growth to monthly and annual precipitation, temperature and aridity between 1991 and 2019. We revealed species-specific responses, namely the (i) recruitment of deep-rooted, heavy-seeded species was positively affected by precipitation in both late winter-early spring and summer, whereas recruitment of shallow-rooted, light-seeded species was weakly influenced by climate fluctuations; (ii) growth of shallow-rooted species was more adversely affected by high summer temperature and drought than the growth of deep-rooted species. The population age structure of all the studied species was affected by the climate of the past decades. Most individuals established in the wet period of the 2000s, fewer in the precipitation-poorer 1990s, and the establishment was considerably reduced in the dry and warm period of the 2010s. Our findings indicate that the change towards warmer and drier climate has a profound effect even on drought-adapted ecosystems such as temperate dry grasslands. However, plant responses to various climatic extremes are species-specific, depending on their characteristics, such as life form or rooting depth. Consequently, the ongoing and anticipated climate warming will likely result in complex changes in species composition and other ecosystem properties of temperate grasslands

ecology↗

bioRxiv2018

Inducible alpha-synuclein overexpression affects human Neural Stem Cells behavior

Converging evidence suggest that levels of alpha-Synuclein (aSyn) expression play a critical role in Parkinsons disease (PD). Several mutations of the SNCA gene, encoding for aSyn have been associated to either the familial or the sporadic forms of PD. Nonetheless, the mechanism underlying wild type aSyn-mediated neurotoxicity in neuronal cells as well as its specific driving role in PD pathogenesis has yet to be fully clarified. In this view, the development of proper in vitro cellular systems is a crucial step.\n\nHere we present a novel human Tet-on hNSC cell line, in which aSyn timing and level of expression can be tightly experimentally tuned. Induction of aSyn in self-renewing hNSCs leads to progressive formation of aSyn aggregates and impairs their proliferation and cell survival. Furthermore, aSyn induction during the neuronal differentiation process results in impaired neurogenic potential due to enhanced refractoriness to exit self-renewal and to increase of gliogenic vs neurogenic competence. Finally, acute aSyn induction in hNSC-derived dopaminergic neuronal cultures results in cell toxicity.\n\nThis novel conditional in vitro cell model system may be a valuable tool for dissecting of aSyn pathogenic effects in hNSCs and neurons and in developing new potential therapeutic strategies.

neuroscience↗

bioRxiv2017

HuD is a neural enhancer of global translation acting on mTORC1-responsive genes and sponged by the Y3 small non-coding RNA

The RNA-binding protein HuD promotes neurogenesis and favors recovery from peripheral axon injury. HuD interacts with many mRNAs, altering both stability and translation efficiency. UV-crosslinking and analysis of cDNA (CRAC) generated a nucleotide resolution map of the HuD RNA interactome in motor neuron-like cells. HuD target sites were identified in 1304 mRNAs, predominantly in the 3UTR, with enrichment for genes involved in protein synthesis and axonogenesis. HuD bound many mRNAs encoding mTORC1-responsive ribosomal proteins and translation factors. Altered HuD expression correlated with the translational efficiency of these mRNAs and overall protein synthesis, in a mTORC1-independent fashion. The predominant HuD target was the abundant, small non-coding RNA Y3, which represented 70% of HuD interaction signal. Y3 functions as a molecular sponge for HuD, dynamically limiting its activity. These findings uncover an alternative route to the mTORC1 pathway for translational control in motor neurons that is tunable by a small non-coding RNA.\n\nGraphical abstract\n\nO_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC=\"FIGDIR/small/205658_ufig1.gif\" ALT=\"Figure 1\">\nView larger version (69K):\norg.highwire.dtl.DTLVardef@17b2d21org.highwire.dtl.DTLVardef@1cfdc8borg.highwire.dtl.DTLVardef@1984f88org.highwire.dtl.DTLVardef@8da1ee_HPS_FORMAT_FIGEXP M_FIG C_FIG

cell biology↗