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Straub, T.

Publications and source records attributed to Straub, T..

At least 19 recordsLinked to original sources

FLRT cell adhesion molecules counteract protocadherin-10-dependent cell segregation during cortical neuron migration

The processes that underlie the folding of the cerebral cortex of large mammals and the evolution of the smooth cortex of modern rodents are incompletely understood. Recent evidence highlighted adhesion-controlled neuronal migration as an important mechanism. Genetic deletion in mice of FLRT1 and FLRT3 cell-adhesion proteins perturbed cortical neuron migration, lead to region-specific neuronal clustering, and introduced cortical folds into the normally smooth mouse cortex, suggesting that Increased expression of FLRT protein may have contributed to cortex smoothening. Here, we identify protocadherin-10 (PCDH10) as a key regulator of neuronal clustering in this context. Using single-cell transcriptomics and spatial proteomics, we describe a cortical neuron subtype in which loss of FLRT1/3 triggers local upregulation of PCDH10 in a specific cortical region. Elevated PCDH10 drives homophilic adhesion and the formation of cell patches that segregate from FLR1/3-deficient migrating neurons. This segregation leads to discrete clustering of Flrt1/3-mutant neurons and correlates with the emergence of cortical folds. Consistent with an evolutionary role, comparative expression analysis showed reciprocal expression of FLRT3 and PCDH10, with high FLRT3 and low PCDH10 expression in the smooth mouse cortex, and the opposite pattern in the developing human cortex. Together, our findings reveal how the antagonism between different classes of cell adhesion molecules affects cortical neuron migration patterns, contributes to region-specific cortical folding and may have promoted evolutionary smoothening of the rodent cortex.

neuroscience↗

Aquarius RNA helicase Protects Pluripotent Stem Cell Identity

Pluripotent stem cells must reconcile rapid replication with a highly dynamic transcriptional program, creating an inherent susceptibility to transcription-replication conflicts (TRCs). We demonstrate that embryonic stem cells (ESCs) operate in a "resilient" replication mode, tolerating high genomic traffic through the constitutive upregulation of R-loop and TRC resolution pathways. Through a targeted functional screen, we identify the RNA helicase Aquarius (AQR) as an essential safeguard of this state. AQR depletion downregulates these resolution factors, collapsing this stress-resistant program and driving ESCs into unstable, heterogenous states, marked by increased transcriptional entropy and cell-to-cell noise. Mechanistically, we show that key identity-defining genes are preferentially located within R-loop and TRC prone regions, making them uniquely vulnerable to AQR depletion. Our findings establish AQR as a critical governor of transcriptional fidelity, demonstrating that genomic resilience is fundamental to maintaining pluripotent cell identity.

cell biology↗

Periweaning diet-induced activation of an IFNγ-mediated regulatory circuit promotes the homeostasis of cytotoxic CD8+ T Cells

Balancing pathogen defence with maintaining tolerance to environmental antigens, such as food or commensals, in neonates is essential for survival and the establishment of life-long immune homeostasis. Instructed by environmental signals type 1 conventional dendritic cells (cDC1) drive either T cell tolerance or immunity. Here, we uncover an interferon (IFN)-{gamma}-driven regulatory circuit in early life that relays dietary cues to spleen cDC1. Loss-of-function demonstrates that IFN{gamma}-mediated STAT1-signaling induces an immunogenic maturation program in spleen cDC1 that instructs cDC1 to expand effector memory CD8 T cells. This program emerges during weaning, when IFN{gamma} production from lymphocytes rises, it occurs in germ-free mice and remains responsive to dietary intervention in adult mice. During the transition from breastfeeding to solid food at weaning, this circuit relays dietary information to spleen cDC1 to shape the effector phenotype of food-antigen specific CD8+ T cells in a feed-forward manner, allowing cDC1 to recalibrate the T cell pool at the moment of nutritional independence.

immunology↗

Diversity of stomatal and cuticular structures affect microbial colonization in temperate forest tree species

Throughout their life cycle, tree leaves are subject to colonization and degradation by microorganisms, including fungi, bacteria, and algae. These relationships co-evolved with chemical properties, leaf shape, and surface structures. Here we quantified leaf abaxial surface texture features based on variables extracted from scanning electron microscopic images, resulting in an abstract quantitative texture complexity score. This complexity score was used to test functional hypotheses such as growth habitat preferences and microbial colonization patterns. Here we show that the evolution of leaf surface texture complexity traits correlated with anatomical features such as stomatal density and leaf orientation and correlated with Ellenberg temperature habitat indicator. Furthermore, increasing leaf surface texture complexity was found to be negatively correlated with plant pathogen richness (broad-leaved species) or lichenization (conifers), suggesting protection effects. Moreover, we found a negative correlation of leaf surface texture complexity with fungal and bacterial specialists. Our results highlight leaf surface texture complexity as a key, previously underappreciated trait shaping microbial diversity and leaf-microbe interaction patterns. This opens promising avenues for future research on plant-microbe co-evolution, trait-based ecosystem modeling, and the potential use of surface traits in forest management and disease resistance strategies.

plant biology↗

Catalytic activity of KMT5B promotes cilia tuft formation without affecting chromatin accessibility of ciliary genes

Multiciliated cells are a specialized cell type found in the brain, reproductive tract and respiratory tract of mammals. Multiciliated cells resembling those of the mammalian lung can also be found on the surface of the epidermis of tadpole stage Xenopus embryos, making the frog an ideal model organism to study this cell type. KMT5B/suv4-20h1 is a histone methyltransferase that writes the dimethyl mark on histone 4, lysine 20 (H4K20). We have previously shown that the multiciliated cells of embryos lacking KMT5B have depleted cilia and a reduced actin cap, and that knockdown of both KMT5B and KMT5C/suv4-20h2 leads to downregulation of ciliogenic genes. Here we further tease out the independent function of KMT5B in multiciliogenesis, and show that single knockdown of KMT5B, but not KMT5C leads to aberrant transcription and the downregulation of cilia genes. This phenotype is ameliorated by overexpression of catalytically active PHF8, an H4K20me1 demethylase, while a hormone inducible variant of multicilin (MCI), the master regulator of cilia tuft formation, has no effect. Notably, the expression of key transcription factors of ciliogenesis is unaffected by KMT5B depletion, and the transcriptional effect of KMT5B depletion dominates the response to ectopic multicilin. Finally, ATAC seq analysis in animal caps reveals that knockdown of KMT5B results globally in very few peaks with differential activity and does not compact chromatin at ciliary genes. Taken together, this suggests that KMT5B regulates multiciliated cells through an alternative pathway to the canonical MCI-driven multiciliogenic program.

developmental biology↗

Organoid-based modeling of platinum resistance identifies KRT17 as both a response mediator and biomarker for targeted therapy in ovarian cancer

Variable platinum responses drive high mortality in high-grade serous ovarian cancer (HGSOC), but to date, there is no molecular approach to define resistance levels for clinical decision-making. Here, we developed the organoid drug resistance assay (ODR-test) with patient-derived organoids from our ovarian cancer biobank and found that all HGSOC patients develop molecular resistance under exposure to carboplatin, although with varying clinical implications. Sustained phenotypic reprogramming and cellular plasticity under carboplatin pressure emerged as a conserved mechanism irrespective of the basal resistance level. Transcriptional and proteomic analyses revealed changes in cell adhesion and differentiation in post-platinum lines as adaptive responses that drive the increase in resistance. We identified Keratin 17 (KRT17) as a mediator of developing platinum resistance and validated its function by CRISPR/Cas9 and overexpression. Additionally, we found that KRT17 expression status (K-score) is a significant negative prognostic histopathological biomarker in a large cohort (N=384) of advanced HGSOC patients. In organoids, increased KRT17 levels enhanced sensitivity to PI3K/Akt inhibitors Alpelisib and Afuresertib, highlighting the potential of KRT17 as a stratification biomarker for targeted therapies.

cancer biology↗

Digital Reprogramming Decodes Epigenetic Barriers of Cell Fate Changes

The fates of differentiated cells in our body can be induced to change by nuclear reprogramming. In this way, cells valuable for therapeutic purposes and disease modeling can be produced. However, the efficiency of this process is low, partly due to properties of somatic donor nuclei which stabilize their differentiated fate but also act as barriers reprogramming-associated cell fate changes. The identity of these reprogramming barriers is not fully understood. Here, we developed an artificial intelligence-based approach to model nuclear reprogramming and used it to identify the chromatin modification H3K27ac as an epigenetic barrier to reprogramming-induced cell fate changes. Using reprogramming by nuclear transfer (NT) to eggs of Xenopus laevis as a model system, we profiled chromatin modifications in differentiated cell types alongside gene expression patterns before and after reprogramming. Our model integrated the data and by leveraging model predictions, we find that genes resisting inactivation during reprogramming display chromatin modification barcodes. This revealed H3K27ac as a novel candidate barrier to NT reprogramming. Reducing H3K27ac levels using p300/CBP inhibitors before reprogramming led to an improved downregulation of genes linked to H3K27ac-modified enhancers after reprogramming. Importantly, these effects were accompanied by improved embryonic development of the resulting nuclear transfer embryos. In summary, our study identified H3K27ac as a safeguarding mechanism of cellular identities and as a reprogramming barrier during NT. Hence, the here-developed Digital Reprogramming" approach is capable of modelling and improving current cell-fate reprogramming strategies.

genomics↗

H3K4 methylation-promoted transcriptional memory ensures faithful zygotic genome activation and embryonic development

In the life of a vertebrate embryo, gene expression is initiated for the first time at zygotic genome activation (ZGA). Maternally expressed transcription factors present in the embryo are essential for this process. However, it is unknown if active chromatin modifications established in the gamete are propagated in the embryo as an epigenetic memory to support ZGA and embryonic development. Here, we provide evidence that in Xenopus laevis, H3K4 methylation provides an epigenetic memory of active chromatin states. We show that this is required for faithful zygotic genome activation and successful embryonic development. Chromatin configurations of promoters displaying high H3K4me3 intensity and breadth, alongside DNA hypomethylation and increased GC content, are propagated from the gametes to the embryo across multiple cell divisions and a transcriptionally quiescent phase in early development. We show that this transmission of H3K4 methylation is essential for precise zygotic genome activation and expression of key pioneer ZGA transcription factors Pou5f3.2 and Sox3. Finally, we demonstrate that the H3K4 methyltransferases Kmt2b and Cxxc1 ensure transcription-independent propagation of H3K4me3 and proper zygotic gene expression. In summary, our study establishes the role of H3K4 methylation in maintaining memory of active chromatin states in Xenopus embryos and reveals its importance for successful embryonic development.

developmental biology↗

Mechanical Centrosome Fracturing during Cell Navigation

The centrosome is the primary microtubule orchestrator in most eukaryotic cells, nucleating and anchoring microtubules that grow radially and exert forces on cargos. At the same time, mechanical stresses from the microenvironment and cellular shape changes compress and bend microtubules. Yet, centrosomes are membrane-less organelles, raising the question of how centrosomes withstand mechanical forces. Here we discover that centrosomes in non-dividing cells can mechanically fracture. We reveal that centrosomes experience mechanical deformations during microenvironmental confinement and navigational pathfinding by motile cells. Coherence of the centrosome is maintained by Dyrk3, preventing fracturing by mechanical forces. Centrosome fracturing impedes cellular function by generating coexisting microtubule organizing centers that compete during path navigation and thereby cause cellular entanglement in the microenvironment. Our findings show that non-dividing cells actively maintain the integrity of the centrosome to withstand mechanical forces. Given that almost all cells in multicellular organism experience forces, these results suggest that centrosome stability preservation is fundamental during development, tissue maintenance, immunology, and disease. One-Sentence SummaryPathfinding during cellular motility causes centrosome breakage counteracted by Dyrk3 activity.

cell biology↗

Interferon-induced activation state of circulating dendritic cells and monocytes triggered by yellow fever vaccination correlates with early protective antibody responses

Yellow fever vaccination provides long-lasting protection and is a unique model for studying the immune response to an acute RNA virus infection in humans. To elucidate the early innate immune events preceding the rapid generation of protective immunity, we performed transcriptome analysis of human blood dendritic cell (DC) and monocyte subpopulations before and 3, 7, 14 and 28 days after vaccination. We detected temporary upregulation of IFN-stimulated genes (ISG) in all DC and monocyte subsets on day 3 and 7 after vaccination as well as cell type specific responses and temporal dynamics. Single cell RNA sequencing revealed rapid appearance of activated DC and monocyte clusters dominated by ISGs, inflammatory chemokines and genes involved in antigen processing and presentation. This was confirmed by flow cytometric analysis in a large cohort of vaccinees. We identified SIGLEC1/CD169 upregulation as a sensitive indicator of the transient IFN-induced activation state elicited in DCs and monocytes by YF17D vaccination correlating with early protective IgM antibody responses.

immunology↗

Neuronal Hyperactivity in Neurons Derived from Individuals with Grey Matter Heterotopia

Periventricular heterotopia (PH), a common form of grey matter heterotopia associated with developmental delay and drug-resistant seizures, poses a challenge in understanding its neurophysiological basis. Human cerebral organoids (hCOs) derived from patients with causative mutations in FAT4 or DCHS1 mimic PH features. However, neuronal activity in these 3D models has not yet been invetigated. Here, silicon probe recordings revealed exaggerated spontaneous spike activity in FAT4 and DCHS1 hCOs, suggesting functional changes in neuronal networks. Transcriptome and proteome analyses identified changes in gene ontology terms associated with neuronal morphology and synaptic function. Furthermore, patch-clamp recordings revealed a decreased spike threshold specifically in DCHS1 neurons, likely due to increased somatic voltage-gated sodium channels. Morphological reconstructions and immunostainings revealed greater morphological complexity of PH neurons and synaptic alterations contributing to hyperactivity, with morphological rescue observed in DCHS1 neurons by wild-type DCHS1 expression. Overall, we provide new comprehensive insights into the cellular changes underlying symptoms of grey matter heterotopia.

neuroscience↗

Dbf4-Dependent Kinase Finetunes INO80 Function at Chromosome Replication Origins

The highly conserved Dbf4-Dependent Kinase (DDK) plays a pivotal role in the nucleus during S phase, where it directly phosphorylates the replicative helicase, the minichromosome maintenance (MCM) complex. This leads to the initiation of chromosome replication. However, aside from the MCM complex, few other targets have been identified to date, leaving DDK an understudied kinase. Here, we describe a two-pronged mass spectrometry-based approach and define the nuclear DDK-dependent phosphoproteome, which consists of approximately 400 phosphorylation events. Within this network, we found that DDK directly phosphorylates the Arp8 subunit of the multi-subunit chromatin remodeler complex INO80. Arp8 phosphorylation stabilises INO80s intramolecular complex integrity, which finetunes its nucleosome spacing activity at replication origins. This adjustment of origin chromatin architecture stimulates replication and is important for the response to replication stress. Our results represent a significant advance in our understanding of the molecular mechanisms underlying the regulation of replication origins.

biochemistry↗

Cortex Folding by Combined Progenitor Expansion and Adhesion-Controlled Neuronal Migration

Folding of the mammalian cerebral cortex into sulcal fissures and gyral peaks is the result of complex processes that are incompletely understood. Previously we showed that genetic deletion of Flrt1/3 adhesion molecules causes folding of the smooth mouse cortex into sulci resulting from increased lateral dispersion and faster neuron migration, without progenitor expansion. Here, we find that combining the Flrt1/3 double knockout with an additional genetic deletion that causes progenitor expansion, greatly enhances cortex folding. Expansion of intermediate progenitors by deletion of Cep83 results in enhanced formation of sulci. Expansion of apical progenitors by deletion of Fgf10 results in enhanced formation of gyri. Single cell transcriptomics and simulations suggest that changes in adhesive properties of cortical neurons, their proportions and densities in the cortical plate, combined with lateral dispersion during their radial migration are important folding parameters. These results identify key developmental mechanisms that cooperate to promote cortical gyrification. HIGHLIGHTSO_LICortex folding is enhanced by combining progenitor expansion and divergent migration. C_LIO_LIConcomitant expansion of intermediate progenitors results in the formation of sulci C_LIO_LIConcomitant expansion of apical progenitors results in the formation of gyri C_LIO_LIProgenitors differentially affect cortical neurons with distinct adhesive properties C_LI

neuroscience↗

Optimizing biomedical information retrieval with a keyword frequency-driven Prompt Enhancement Strategy

BackgroundMining the enormous pool of biomedical literature to extract accurate responses and relevant references is a daunting challenge, made more difficult by the domains interdisciplinary nature, specialized jargon, and continuous evolution. Early natural language processing (NLP) approaches faced considerable hurdles in comprehending the nuances of natural language which frequently led to false answers. However, the discovery of transformer models has immensely boosted the cause by enabling researchers to create larger and more potent language models known as large language models (LLMs). These LLMs have opened up new possibilities for enhancing question-answering (QA) tasks. Even with these technological advances, the current LLM-based solutions for querying specialized domains like biology and biomedicine still have issues in generating up to date responses while preventing "hallucination" or the generation of plausible but factually incorrect responses. ResultsThis paper focuses on prompt enhancement using retrieval-augmented architecture as a strategy to guide LLMs towards generating more meaningful responses for biomedical question-answering tasks. We evaluated two prompt enhancement approaches using GPT-3 and GPT-4, examining their effectiveness in retrieving relevant information from complex landscape of biomedical literature. Our proposed approach leverages explicit signals in user queries to extract meaningful contexts from a vast pool of information, addressing the shortcomings of traditional text embedding-based prompt enhancement methods. We developed a QA bot WeiseEule (https://github.com/wasimaftab/WeiseEule-LocalHost) utilizing these prompt enhancement methods and allowing for comparative analysis of their performances. ConclusionsOur findings highlight the importance of prompt enhancement methods that utilize explicit signals in users query over traditional text embedding based counterparts to improve LLM-generated responses in specialized domains such as biology and biomedicine. By providing users complete control over the information that goes into the LLM, our approach tackles some of the major drawbacks of existing web-based chatbots and LLM-based QA systems including hallucinations and the generation of irrelevant or outdated responses.

bioinformatics↗

Genetically- and spatially-defined basolateral amygdala neurons control food consumption and social interaction

The basolateral amygdala (BLA) contains discrete neuronal circuits that integrate positive or negative emotional information and drive the appropriate innate and learned behaviors. Whether these circuits consist of genetically-identifiable and anatomically segregated neuron types, is poorly understood. Also, our understanding of the response patterns and behavioral spectra of genetically-identifiable BLA neurons is limited. Here, we classified 11 glutamatergic BLA cell clusters and found that several of them were anatomically segregated in lateral versus basal amygdala, and anterior versus posterior regions of the BLA. Two of these BLA subpopulations innately responded to valence-specific, whereas one responded to mixed - aversive and social - cues. Positive-valence BLA neurons promoted normal feeding, while mixed selectivity neurons promoted fear learning and social interactions. These findings enhance our understanding of cell type diversity and spatial organization of the BLA and the role of distinct BLA populations in representing valence-specific and mixed stimuli. HighlightsO_LIClassification of molecularly-defined glutamatergic neuron types in mouse BLA with distinct spatial expression patterns. C_LIO_LIBLALypd1 neurons are positive-valence neurons innately responding to food and promoting normal feeding. C_LIO_LIBLAEtv1 neurons innately represent aversive and social stimuli. C_LIO_LIBLAEtv1 neurons promote fear learning and social interactions. C_LI

neuroscience↗

Bruno 1 regulates cytoskeleton dynamics and a temporal splicing transition to promote myofibril assembly, growth and maturation in Drosophila flight muscle

Muscles undergo developmental transitions in gene expression and alternative splicing that are necessary to refine sarcomere structure and contractility. CUG-BP and ETR-3-like (CELF) family RNA binding proteins are important regulators of RNA processing during myogenesis that are misregulated in diseases such as myotonic dystrophy (DM1). Here we report a conserved function for Bruno 1 (Bru1, Arrest), a CELF1/2 family homolog in Drosophila, during early muscle myogenesis. Loss of Bru1 in flight muscles results in disorganization of the actin cytoskeleton leading to aberrant myofiber compaction and defects in pre-myofibril formation. Temporally-restricted rescue and RNAi knockdown demonstrate that early cytoskeletal defects interfere with subsequent steps in sarcomere growth and maturation. Early defects are distinct from a later requirement for bru1 to regulate sarcomere assembly dynamics during myofiber maturation. We identify an imbalance in growth in sarcomere length and width during later stages of development as the mechanism driving abnormal radial growth, myofibril fusion and the formation of hollow myofibrils in bru1 mutant muscle. Molecularly, we characterize a genome-wide transition from immature to mature sarcomere gene isoform expression in flight muscle development that is blocked in bru1 mutants. We further demonstrate that temporally restricted Bru1 rescue can partially alleviate hypercontraction in late pupal and adult stages, but it cannot restore myofiber function or correct structural deficits. Our results reveal the conserved nature of CELF function in regulating cytoskeletal dynamics in muscle development, and demonstrate that defective RNA processing due to misexpression of CELF proteins causes wide-reaching structural defects and progressive malfunction of affected muscles that cannot be rescued by late-stage gene replacement.

developmental biology↗

Single molecule MATAC-seq reveals key determinants of DNA replication origin efficiency

SummaryStochastic origin activation gives rise to significant cell-to-cell variability in the pattern of genome replication. The molecular basis for heterogeneity in efficiency and timing of individual origins is a long-standing question. Here, we developed Methylation Accessibility of TArgeted Chromatin domain Sequencing (MATAC-Seq) to determine single-molecule chromatin accessibility of specific genomic loci after targeted purification in their native chromatin context. Applying MATAC-Seq to selected early-efficient (EE) and late-inefficient (LI) budding yeast replication origins revealed large heterogeneity of chromatin states. Disruption of INO80 or ISW2 chromatin remodeling complexes leads to changes at individual nucleosomal positions that correlate with changes in their replication efficiency. We found a chromatin state with an optimal 100-115bp nucleosome-free region in combination with surrounding well-positioned nucleosomes and open +2 linker region is a strong predictor for efficient origin activation. Thus, MATAC-Seq identifies the large spectrum of alternative chromatin states that co-exist on a given locus previously masked in population-based experiments and provides a mechanistic basis for origin activation heterogeneity during DNA replication of eukaryotic cells. Consequently, our single-molecule assay for chromatin accessibility will be ideal to define single-molecule heterogeneity across many fundamental biological processes such as transcription, replication, or DNA repair in vitro and ex vivo.

biochemistry↗

Plasmacytoid dendritic cells regulate megakaryocyte and platelet homeostasis

Platelet homeostasis is essential for vascular integrity and immune defense. While the process of platelet formation by fragmenting megakaryocytes (thrombopoiesis) has been extensively studied, the cellular and molecular mechanisms required to constantly replenish the pool of megakaryocytes by their progenitor cells (megakaryopoiesis) remains unclear. Here we use intravital 2 photon microscopy to track individual megakaryopoiesis over days. We identify plasmacytoid dendritic cells (pDCs) as crucial bone marrow niche cells that regulate megakaryopoiesis. pDCs monitor the bone marrow for platelet-producing megakaryocytes and deliver IFN- to the megakaryocytic niche to trigger local on-demand proliferation of megakaryocyte progenitors. This fine-tuned coordination between thrombopoiesis and megakaryopoiesis is crucial for megakaryocyte and platelet homeostasis in steady state and stress. However, uncontrolled pDC function within the megakaryocytic niche is detrimental. Accordingly, we show that pDCs activated by SARS-CoV2 drive inappropriate megakaryopoiesis associated with thrombotic complications. Together, we uncover a hitherto unknown megakaryocytic bone marrow niche maintained by the constitutive delivery of pDC-derived IFN-.

immunology↗