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Biology subjects

Simon, E.

Publications and source records attributed to Simon, E..

At least 19 recordsLinked to original sources

Drosophila male genitalia rotation depends on permissive remodeling of the posterior abdomen

One of the most characteristic morphogenetic processes in Drosophila is the 360{degrees} rotation of the male pupal genital disc. This movement is driven by the myosin Myo1D, whose expression in the genital disc is controlled by the Hox gene Abdominal-B. The rotation takes place in contact and relative to the posterior abdomen, yet the contribution of abdominal tissues has remained unclear. Here we show that normal genital disc circumrotation requires active remodeling of posterior abdominal larval epidermal cells that contact the rotating terminalia. Preventing apoptosis in these cells, or increasing EGFR signaling, delays their extrusion and results in incomplete rotation without altering rotational chirality. In parallel, elimination of Extracellular Matrix by Metalloproteinase 1 in these cells, although without leading to their extrusion, is also strictly required for genital disc circumrotation. Inhibition of this metalloproteinase activity leads to persistence of collagen IV and incomplete rotation, revealing an independent requirement for Extracellular Matrix clearance at the disc-abdomen interface. By contrast, genetic conditions that prevent formation or elimination of the male A7 segment do not necessarily impair genital disc rotation, demonstrating that A7 suppression and circumrotation are separable processes. These findings identify posterior abdominal tissue remodeling as an essential extrinsic requirement that enables genital disc circumrotation.

developmental biology↗

Gradient boosting regression and convolution improve deconvolution of bulk transcriptomes

Bulk cell type deconvolution aims to estimate cell type composition from bulk transcriptomic data. So-called pseudobulk simulation, where single-cell RNA-seq data is aggregated to bulk-like expression profiles, represents a central concept in training and testing of deconvolution tools. However, deconvolution methods often lack interpretability and struggle to generalize from simulated to real data. We present GrooD (GradientBoostedDeconvolution), a second-generation deconvolution tool that uses gradient boosted trees trained on pseudobulks from scRNA-seq references. GrooDs pseudobulk simulations account for donor and condition variability to better model the complexity of real-world transcriptomes. We show that GrooD achieves state-of-the-art and superior deconvolution performance on human blood biospecimen. Furthermore, Grood provides visualizations of feature loadings and deconvolution results, that allow mechanistic insights for biological interpretation. We further integrate a convolution framework to assess the transcriptomic similarity between bulk and pseudobulk data, showing that higher similarity can indicate better deconvolution performance. GrooD deconvolution of blood transcriptomes from a large sepsis patient cohort identifies meaningful shifts in immune cell type composition that are associated with disease severity. By combining interpretability, robustness, and heterogeneous pseudobulk simulation, GrooD represents a powerful, user-friendly second-generation tool for cell type deconvolution.

bioinformatics↗

Nucleoside Binding by a Surface Lipoprotein Governs Conjugative ICE Acquisition in Ruminant Mycoplasmas

Integrative and conjugative elements (ICEs) are major mediators of horizontal gene transfer (HGT) in bacteria. However, the role of recipient cells in their acquisition has received little attention. Using the ruminant pathogens Mycoplasma agalactiae and Mycoplasma bovis as minimal models, we combined genome-wide transposon mutagenesis with high-throughput mating assays to identify recipient factors required for ICE acquisition. The surface lipoprotein P48 emerged as the primary determinant of ICE uptake in both species. Structural and functional analyses revealed that P48 is the substrate-binding component of an ABC transporter with nucleoside-binding capacity. A single point mutation that abolished nucleoside binding drastically reduced ICE acquisition, demonstrating that P48-mediated nucleoside recognition is essential for conjugative transfer. However, ICE uptake did not require nucleoside transport, as inactivation of the transporter permease blocked nucleoside analog toxicity but not ICE invasion. Loss of P48 also triggered transcriptional activation of vestigial ICE genes, suggesting that surface recognition affects the intracellular state of the recipient. Remarkably, ICE transmission from recipient-derived donors was unaffected by P48 loss, underscoring its acquisition-specific role. Together, these results reveal a previously unrecognized, surface-exposed recipient factor critical for efficient ICE transfer in mycoplasmas and identify nucleotide binding as a central function in conjugation. By demonstrating that recipient-encoded functions can directly control ICE dissemination, this work challenges the donor-centric paradigm of bacterial conjugation and suggests new strategies to restrict horizontal gene flow in pathogenic and synthetic mycoplasmas. IMPORTANCEIntegrative and conjugative elements (ICEs) are mobile DNA elements that drive bacterial conjugation, a major process by which bacteria exchange genes. Although conjugation has been studied for decades, the focus has been almost exclusively on donor cells and the ICE itself, leaving the role of recipient cells largely overlooked. Using the wall-less ruminant pathogens Mycoplasma agalactiae and Mycoplasma bovis as minimal models, we discovered that a single recipient lipoprotein is required for efficient ICE uptake. Our data show that nucleoside recognition by P48, but not transport, is critical for conjugation, revealing an unexpected mechanistic link between nutrient sensing and gene acquisition. These findings shift the paradigm of conjugation from a donor-driven process to one jointly determined by donor and recipient functions. By identifying a recipient-encoded determinant of ICE transfer, this work opens new avenues to control horizontal gene flow in both pathogenic and engineered bacteria. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=151 SRC="FIGDIR/small/677790v1_ufig1.gif" ALT="Figure 1"> View larger version (26K): org.highwire.dtl.DTLVardef@f7cc15org.highwire.dtl.DTLVardef@77a34eorg.highwire.dtl.DTLVardef@82699corg.highwire.dtl.DTLVardef@1e3be93_HPS_FORMAT_FIGEXP M_FIG C_FIG

microbiology↗

Strengths of relationships among soil microbial and organic matter properties are scale-dependent

Relationships among variables in ecological systems are inherently scale-dependent, especially in heterogeneous systems. Yet it remains to be examined whether relationships among variables vary across observation scales in soil. Generally, it is desirable that observation scale matches the intrinsic scale of a process or pattern. Millimetre-sized soil aggregates are closer to the intrinsic scale of microbial communities than traditionally studied bulk soil samples, making them more suitable for studying potential links between microbial communities and their environment. To explore the effect of observation scale on relationships among soil parameters, we measured bacterial, archaeal, and fungal taxa richness and density, organic matter properties (e.g., carbon and nitrogen content, stoichiometric and isotopic ratios), and soil water content in individual aggregates and aliquots of homogenised soil cores, bulk soil samples, in two soil layers. We analysed pairwise correlations among these variables and assessed whether individual aggregates systematically differed from bulk soil samples. Organic matter properties were more strongly correlated in bulk soil samples, consistent with the idea that increasing the sample volume reduces noise. In contrast, microbial community and organic matter properties showed weaker correlations in bulk soil samples than aggregates in topsoil. In addition, we found that aggregates and bulk soil samples differed systematically in individual microbial and organic matter properties, particularly in the topsoil. Our study demonstrates that relationships among variables in soil are spatial scale-dependent. Aggregates offer valuable insights into microbial communities in soil, complementing bulk soil samples, and are useful for studying links between microbial communities and their environment.

ecology↗

Model substrate particles uncover dynamics of microbial communities associated with particulate organic matter decomposition in soil

Soil organic matter is the largest terrestrial reservoir of organic carbon. Its particulate fraction, particulate organic matter (POM), serves as a resource and surface for microbial colonization. Degradation of complex biopolymers like cellulose and chitin requires extracellular enzymes produced by phylogenetically diverse microbes. Despite their importance for carbon cycling, the structure and spatio-temporal dynamics of POM-associated microbial communities in soil and how specific substrates influence them remain poorly understood. This study investigated whether microbial communities associated with POM change in composition and richness over time and whether chitin and cellulose select for distinct fungal and bacterial taxa. We incubated self-manufactured, millimetre-sized model substrate particles containing chitin or cellulose in soil under laboratory and field conditions. We assessed particle-associated communities at multiple time points over a 50-day-long incubation in the lab and after 47 days in the field. Our results show that community structure and temporal dynamics of particle-associated microbial communities were substrate-specific. While microbial biomass increased on both particle types, chitin-associated microbial communities exhibited stronger temporal changes. Communities on chitin and cellulose particles were enriched in specific bacterial and fungal genera compared to communities in the surrounding soil. We demonstrate that microbial communities associated with model chitin particles underwent notable temporal changes, including decreased microbial richness and shifts in community composition over the incubation period. This study shows the potential of model particles to advance our understanding of particle- and substrate-associated microbial communities in soil.

microbiology↗

Bacterial Conjugation in the Ruminant Pathogen Mycoplasma agalactiae is Influenced by Eukaryotic Host Factors

Bacterial conjugation plays a pivotal role in the evolution and adaptation of genome-reduced mycoplasmas. Despite their fast evolution rate, the conjugative properties of these organisms remain largely understudied, particularly in vivo. In the present study, the ruminant pathogen Mycoplasma agalactiae was used as a model organism to document the conjugative properties of mycoplasmas in environments of increasing complexity, from axenic to cell and organotypic culture conditions. Compared to axenic mating conditions, mycoplasma co-cultivation with goat epithelial cells or bovine precision-cut lung slices (PCLS) resulted in enhanced mating frequencies with high rates of M. agalactiae Integrative and Conjugative Element (ICEA) self-dissemination. These results were conditioned by the presence of eukaryotic cells in the culture and influenced by competition between mating partners but were not limited to M. agalactiae, as similar results were observed with Mycoplasma bovis. Mycoplasma conjugation ex vivo was further characterized by analyzing mycoplasma chromosomal transfer (MCT), a newly discovered mechanism of horizontal exchange of chromosomal DNA that generates mosaic genomes. Although closely associated with ICEA transfer, MCT was detected at low rates under cell and organotypic culture conditions suggesting a complex interplay between these two conjugative processes or a poor viability of the MCT progeny. Finally, mating experiments under nutrient-deprived conditions identified nucleotide stress as a potential factor influencing the modulation of mycoplasma conjugation by eukaryotic host cells. In conclusion, these results suggest that horizontal gene transfer in vivo is likely underestimated and provide valuable models to further studying mycoplasma conjugation ex vivo. IMPORTANCEConjugation is an evolutionary shortcut that bacteria use to exchange genetic information with their neighbors. Despite the fast evolution rate of the genome-reduced mycoplasmas, their conjugative properties remain largely understudied, particularly in vivo. Here we used the ruminant pathogen Mycoplasma agalactiae to study how mycoplasmas conjugate in co-culture with hosts-derived cells and tissues. Interestingly, conjugation was stimulated when mycoplasmas were co-cultured with eukaryotic cells. This was documented by monitoring the self-propagation of a mobile genetic element known as Integrative and Conjugative Element (ICE) and the exchange of chromosomal DNA leading to the formation of mosaic genomes. While ICE transfer was observed at high frequency, only a few mosaic genomes were detected in the presence of eukaryotic cells. Further data point towards nucleotide stress as a possible factor modulating mycoplasma conjugation in cellular environments. These results suggest that mycoplasma-host interactions may stimulate conjugation in vivo.

microbiology↗

InterPLM: Discovering Interpretable Features in Protein Language Models via Sparse Autoencoders

AO_SCPLOWBSTRACTC_SCPLOWProtein language models (PLMs) have demonstrated remarkable success in protein modeling and design, yet their internal mechanisms for predicting structure and function remain poorly understood. Here we present a systematic approach to extract and analyze interpretable features from PLMs using sparse autoencoders (SAEs). By training SAEs on embeddings from the PLM ESM-2, we identify thousands of human-interpretable latent features per layer that highlight hundreds of known biological concepts such as binding sites, structural motifs, and functional domains. In contrast, examining individual neurons in ESM-2 reveals significantly less conceptual alignment, suggesting that PLMs represent most concepts in superposition. We further demonstrate that a larger PLM (ESM-2 with 650M parameters) captures substantially more interpretable concepts than a smaller PLM (ESM-2 with 8M parameters). Beyond capturing known annotations, we show that ESM-2 learns coherent concepts that do not map onto existing annotations and propose a pipeline using language models to automatically interpret novel latent features learned by the SAEs. As practical applications, we demonstrate how these latent features can fill in missing annotations in protein databases and enable targeted steering of protein sequence generation. Our results demonstrate that PLMs encode rich, interpretable representations of protein biology and we propose a systematic framework to uncover and understand these latent features. In the process, we recover both known biology and potentially new protein motifs. As community resources, we introduce InterPLM (interPLM.ai), an interactive visualization platform for investigating learned PLM features, and release code for training and analysis at github.com/ElanaPearl/interPLM.

bioinformatics↗

Filopodia-like Structures are Essential for Steroid Release

Steroid hormones, crucial for development and physiology, were traditionally believed to diffuse passively through membranes. However, recent evidence shows insect steroid ecdysone being secreted via regulated exocytosis, but the mechanisms ensuring successful hormone release into circulation remain unclear. Our study identifies specialized membrane protrusions, filopodia-like structures named "hormonemes," in the Drosophila prothoracic gland as essential for steroid vesicle release. Confocal imaging reveals that these actin- and tubulin-rich structures form a membrane-intertwined basal domain critical for secretion. Disrupting filopodia by interfering with BM interactions--perlecan or {beta}-integrin--or filopodia-specific protein expression---actinin--significantly reduces ecdysone signaling by impairing its release, despite proper production in the gland. Additionally, filopodia dynamics, such as length and density, align with secretion timing and hormone circulating levels, suggesting their role in synchronizing release with physiological needs. The systematic presence of membrane protrusions in steroid-secreting glands across species prompts a comprehensive re-evaluation of steroid release mechanisms.

developmental biology↗

Distinct microbial communities are linked to organic matter properties in millimetre-sized soil aggregates

Soils provide essential ecosystem services and represent the most diverse habitat on Earth. It has been suggested that the presence of various physico-chemically heterogenous microhabitats supports the enormous diversity of microbial communities in soil. However, little is known about the relationship between microbial communities and their immediate environment at the micro- to millimetre-scale. In this study, we examined whether bacteria, archaea, and fungi organise into distinct communities in individual 2-millimetre-sized soil aggregates and compared them to communities of homogenized bulk soil samples. Furthermore, we investigated their relationship to their local environment by concomitantly determining microbial community structure and physico-chemical properties from the same individual aggregates. Aggregate-communities displayed exceptionally high beta-diversity, with 3-4 aggregates collectively capturing more diversity than their homogenized parent soil core. Up to 20-30% of ASVs (particularly rare ones) were unique to individual aggregates selected within a few centimetres. Aggregates and bulk soil samples showed partly different dominant phyla, indicating that taxa that are potentially driving biogeochemical processes at the small scale may not be recognized when analysing larger soil volumes. Microbial community composition and richness of individual aggregates were closely related to aggregate-specific carbon and nitrogen content, carbon stable-isotope composition, and soil moisture, indicating that aggregates provide a stable environment for sufficient time to allow co-development of communities and their environment. We conclude that the soil microbiome is a metacommunity of variable subcommunities. Our study highlights the necessity to study small, spatially coherent soil samples to better understand controls of community structure and community-mediated processes in soils.

ecology↗

In Silico Treatment: a computational framework for animal model selection and drug assessment

The translation of findings from animal models to human disease is a fundamental part in the field of drug development. However, only a small proportion of promising preclinical results in animals translate to human pathophysiology. This underscores the necessity for novel data analysis strategies to accurately evaluate the most suitable animal model for a specific purpose, ensuring cross-species translatability. To address this need, we present In Silico Treatment (IST), a computational method to assess translation of disease-related molecular expression patterns between animal models and humans. By simulating changes observed in animals onto humans, IST provides a holistic picture of how well animal models recapitulate key aspects of human disease, or how treatments transform pathogenic expression patterns to healthy ones. Furthermore, IST highlights particular genes that influence molecular features of pathogenesis or drug mode of action. We demonstrate the potential of IST with three applications using bulk transcriptomics data. First, we assessed two mouse models for idiopathic pulmonary fibrosis (IPF): one involving injury with intra-tubular Bleomycin exposure, and the other Adeno-associated-virus-induced, TGF{beta}1-mediated tissue transformation (AAV6.2-TGF{beta}1). Both models exhibited gene expression patterns resembling extracellular matrix derangement in human IPF, whereas differences in VEGF-driven vascularization were observed. Second, we confirmed known features of non-alcoholic steatohepatitis (NASH) mouse models, including choline-deficient, l-amino acid-defined diet (CDAA), carbon tetrachloride hepatotoxicity injury (CCl4) and bile duct ligation surgery (BDL). Overall, the three mouse models recapitulated expression changes related to fibrosis in human NASH, whereas model-specific differences were found in lipid metabolism, inflammation, and apoptosis. Third, we reproduced the strong anti-fibrotic signature and induction of the PPAR signaling observed in the Elafibranor experimental treatment for NASH in the CDAA model. We validated the contribution of known disease-related genes to the findings made with IST in the IPF and NASH applications. The complete data integration IST framework, including an interactive app to integrate and compare datasets, is made available as an open-source R package. Author summaryPreclinical testing plays a pivotal role in the drug development process, serving as a crucial evaluation phase before a new drug can be tested on humans in clinical trials. The drug must undergo a rigorous evaluation in in vivo and in vitro preclinical studies to assess its safety and efficacy. However, positive outcomes in preclinical animal models do not always translate to positive results in humans, mainly due to biological differences. Therefore, selecting an animal model that closely mirrors human disease traits and detecting and accounting for model limitations is of paramount importance. Over the last decade, the availability of gene expression data in both animals and humans has substantially increased. Gene expression states and perturbations are routinely employed as a proxy to predict and understand changes in disease states. Here, we developed In Silico Treatment, a computational method designed to overlay the gene expression changes observed in animals onto humans, quantifying the change in human disease status. We applied this method to mouse models for idiopathic pulmonary fibrosis and non-alcoholic steatohepatitis, two severe fibrotic diseases. We successfully identified known features of the disease models and provide a granular gene-level rationale behind our predictions. Consequently, our method shows promise as an effective approach to improve animal model selection and thus clinical translation.

bioinformatics↗

Assessment of the level of pollution based on soil and Tilia europaea leaves

Anthropic activities related to urbanization release pollutants, including potentially toxic elements, into the atmosphere and these are eventually deposited in soils, water, infrastructures, vegetation. Urban soil and plant leaves are widely used as ecological indicators to assess the effect of urbanization. This study aimed to evaluate the impact of urbanization based on the elemental concentration of soil and leaves of Tilia x europaea from urban, suburban, and rural areas along an urbanization gradient in Debrecen, Hungary. Using the ICP-OES technique, we measured the concentration of Ba, Cd, Co, Cr, Cu, Ni, Pb, Sr, and Zn and based on the measured concentration, bioaccumulation factor (BAF) was calculated. The highest concentration of all elements was found in soil samples from urban areas, with an increasing tendency along the urbanization gradient. A significant difference was found along the urbanization gradient only for Cr based on the plant leaves. T. x europaea showed bioaccumulation capacity for Cr and Sr. Our results suggest that urbanization has a remarkable effect on the elemental concentration of soil, which is a perfect ecological indicator. At the same time, we concluded that the T. x europaea was not sensitive enough to indicate the effect of urbanization.

ecology↗

Transcriptomic signatures of progressive and regressive liver fibrosis and portal hypertension

Persistent liver injury triggers a fibrogenic program that causes pathologic remodelling of the hepatic microenvironment (i.e., liver fibrosis) and portal hypertension. The dynamics of gene regulation during liver disease progression and regression remain understudied. Here, we generated hepatic transcriptome profiles in two well-established liver disease models at peak fibrosis and during spontaneous regression after the removal of the inducing agents. We linked the dynamics of key liver disease readouts, such as portal pressure, collagen proportionate area, and transaminase serum levels, to most differentially expressed genes, enabling the identification of transcriptomic signatures of progressive vs. regressive liver fibrosis and portal hypertension. These candidate biomarkers (e.g., Scube1, Tcf4, Src, Hmga1, Trem2, Mafk, Mmp7) were also validated in RNA-seq datasets of patients with cirrhosis and portal hypertension. Finally, deconvolution analysis identified major cell types and suggested an association of macrophage and portal hepatocyte signatures with portal hypertension and fibrosis area in both models.

systems biology↗

Assessment of environmental impacts based on particulate matter, and chlorophyll content of urban trees

Trees improve air quality, and they have irreplaceable aesthetic value in urban landscapes. The amount of dust deposited on tree leaves is a simple and cost-effective indicator of air quality. Our aim was to explore particle filtering capacity of roadside trees in an urbanized area. We also assessed the impact of meteorological conditions on the amount of deposited dust. We measured the leaf surface deposition, and chlorophyll content of leaves along a road section that started at an intersection and ended in a less disturbed suburban area in Debrecen city, Hungary. Samples were collected in July, and September from Celtis occidentalis, a frequent species. We found a significant negative correlation between dust deposition on the leaves, and the distance from the intersection in July, meaning that the amount of dust on the leaves decreased as the distance from traffic increased. In September, dust deposition decreased considerably compared to July, caused by the rainfall before the second sampling. Chlorophyll content also had a significant negative correlation with the distance from the traffic intersection in July, as it decreased towards the less disturbed end of the transect. We also found a positive correlation between dust deposition and chlorophyll content in July. Surprisingly, the exposure to moderate amounts of pollutants in the air caused an increase in chlorophyll content. Our findings suggest that dust deposition on leaves serves as a reliable indicator of traffic intensity, because excess dust caused by the proximity of car traffic can be detected on the leaf surface. Although, certain weather conditions like rainfall and wind can disrupt the patterns in dust deposition that have developed over an extended period through wash-off and resuspension. Hence, it is advisable to consider these effects while selecting the sampling time and evaluating the results. HighlightsO_LILeaves of urban trees are used as bioindicators of deposited dust pollution. C_LIO_LIDust deposition, and chlorophyll content was measured along a roadside transect. C_LIO_LIWe found that dust and chlorophyll content decreased with distance from traffic emissions. C_LIO_LIModerate level of dust pollution resulted in an increase in chlorophyll content. C_LI

ecology↗

From Diversity to Complexity: Microbial Networks in Soils

Network analysis has been used for many years in ecological research to analyze organismal associations, for example in food webs, plant-plant or plant-animal interactions. Although network analysis is widely applied in microbial ecology, only recently has it entered the realms of soil microbial ecology, shown by a rapid rise in studies applying co-occurrence analysis to soil microbial communities. While this application offers great potential for deeper insights into the ecological structure of soil microbial ecosystems, it also brings new challenges related to the specific characteristics of soil datasets and the type of ecological questions that can be addressed. In this Perspectives Paper we assess the challenges of applying network analysis to soil microbial ecology due to the small-scale heterogeneity of the soil environment and the nature of soil microbial datasets. We review the different approaches of network construction that are commonly applied to soil microbial datasets and discuss their features and limitations. Using a test dataset of microbial communities from two depths of a forest soil, we demonstrate how different experimental designs and network constructing algorithms affect the structure of the resulting networks, and how this in turn may influence ecological conclusions. We will also reveal how assumptions of the construction method, methods of preparing the dataset, and definitions of thresholds affect the network structure. Finally, we discuss the particular questions in soil microbial ecology that can be approached by analyzing and interpreting specific network properties. Targeting these network properties in a meaningful way will allow applying this technique not in merely descriptive, but in hypothesis-driven research.

ecology↗

Glypicans specifically regulate Hedgehog signaling through their interaction with Ihog in cytonemes

The conserved family of Hedgehog (Hh) signaling proteins plays a key role in cell-cell communication in development, tissue repair and cancer progression. These proteins can act as morphogens, inducing responses dependent on the ligand concentration in target cells located at a distance. Hh proteins are lipid modified and thereby have high affinity for membranes, which hinders the understanding of their spreading across tissues. Direct contact between cell membranes by filopodia-like structures (also known as cytonemes) could be the simplest explanation for Hh dispersal. To better understand this signaling mechanism, we have analyzed in Drosophila the interaction between the glypicans that, besides for other pathways, are necessary for Hh signaling, plus the adhesion molecules and Hh coreceptors Ihog and Boi. We describe that glypicans (Dally and Dally-like protein) are required to maintain Ihog, but not Boi, protein levels. We also show that ectopic Ihog stabilizes cytonemes through its interaction with glypicans, and we determine that two Ihog fibronectin III domains are essential for this interaction. Our data suggest that this interaction with Ihog in cytonemes confers the specificity of glypicans for Hh signaling.

developmental biology↗

Intranasal Immunization with a Lentiviral Vector Coding for SARS-CoV-2 Spike Protein Confers Vigorous Protection in Pre-Clinical Animal Models

To develop a vaccine candidate against COVID-19, we generated a Lentiviral Vector (LV), eliciting neutralizing antibodies against the Spike glycoprotein of SARS-CoV-2. Systemic vaccination by this vector in mice, in which the expression of the SARS-CoV-2 receptor hACE2 has been induced by transduction of respiratory tract cells by an adenoviral vector, conferred only partial protection, despite an intense serum neutralizing activity. However, targeting the immune response to the respiratory tract through an intranasal boost with this LV resulted in > 3 log10 decrease in the lung viral loads and avoided local inflammation. Moreover, both integrative and non-integrative LV platforms displayed a strong vaccine efficacy and inhibited lung deleterious injury in golden hamsters, which are naturally permissive to SARS-CoV-2 replication and restitute the human COVID-19 physiopathology. Our results provide evidence of marked prophylactic effects of the LV-based vaccination against SARS-CoV-2 and designate the intranasal immunization as a powerful approach against COVID-19. HighlightsA lentiviral vector encoding for Spike predicts a promising COVID-19 vaccine Targeting the immune response to the upper respiratory tract is key to protection Intranasal vaccination induces protective mucosal immunity against SARS-CoV-2 Lung anti-Spike IgA responses correlate with protection and reduced inflammation

immunology↗

Inhibition of IL34 unveils tissue-selectivity and is sufficient to reduce microglial proliferation in chronic neurodegeneration

The proliferation and activation of microglia, the resident macrophages in the brain, is a hallmark of many neurodegenerative diseases such as Alzheimers disease (AD) and prion disease. Colony stimulating factor 1 receptor (CSF1R) is critically involved in regulating microglial proliferation, and CSF1R blocking strategies have been recently used to modulate microglia in neurodegenerative diseases. However, CSF1R is broadly expressed by many cellular types and the impact of its inhibition on the innate immune system is still unclear. CSF1R can be activated by two independent ligands, CSF1 and interleukin 34 (IL-34). Recently, it has been reported that microglia development and maintenance depend on IL-34 signalling. In this study, we evaluate the inhibition of IL-34 as a novel strategy to reduce microglial proliferation in neurodegenerative diseases, using the ME7 model of prion disease. Selective inhibition of IL-34 showed no effects on peripheral macrophages populations in healthy mice, avoiding the side effects observed after CSF1R inhibition on the systemic compartment. However, we observed a reduction in microglial proliferation after IL-34 inhibition in prion-diseased mice, indicating that microglia could be more specifically targeted by reducing IL-34 and that this ligand plays an important role in the modulation of microglia population during neurodegeneration. Overall, our results suggest that control of microglial response through IL-34 blockade could be a potential therapeutic approach in neurodegenerative diseases.

neuroscience↗

Accelerating Protein Design Using Autoregressive Generative Models

The ability to design functional sequences and predict effects of variation is central to protein engineering and biotherapeutics. State-of-art computational methods rely on models that leverage evolutionary information but are inadequate for important applications where multiple sequence alignments are not robust. Such applications include the prediction of variant effects of indels, disordered proteins, and the design of proteins such as antibodies due to the highly variable complementarity determining regions. We introduce a deep generative model adapted from natural language processing for prediction and design of diverse functional sequences without the need for alignments. The model performs state-of-art prediction of missense and indel effects and we successfully design and test a diverse 105-nanobody library that shows better expression than a 1000-fold larger synthetic library. Our results demonstrate the power of the alignment-free autoregressive model in generalizing to regions of sequence space traditionally considered beyond the reach of prediction and design.

systems biology↗