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Stappert, D.

Publications and source records attributed to Stappert, D..

5 recordsLinked to original sources

The core herpes simplex-1 fusion complex drives cell-to-cell spreading of pathological Tau

Neurodegenerative diseases such as Alzheimers disease (AD) are characterized by the pathological aggregation of the host-encoded protein Tau into amyloid fibrils. Pathologic protein aggregates composed of Tau are able to spread from cell-to-cell, thereby contributing to disease progression. The exact mechanisms of intercellular dissemination remain ill-defined. Mounting evidence links the herpes simplex virus-1 (HSV-1) to the aetiology of AD. HSV-1 is a prevalent neurotropic virus that replicates in epithelial cells at the site of infection, and subsequently establishes lifelong latency in the peripheral and central nervous systems. In the quiescent latent state, AD brains show elevated expression of structural viral proteins in the absence of virus production. We hypothesized that HSV-1 glycoproteins involved in viral attachment and fusion facilitate the intercellular spreading of proteopathic seeds. Using cellular models, we demonstrate that expression of the HSV-1 core fusion complex, essential for viral entry and direct cell-to-cell transmission, is sufficient to promote aggregate dissemination between cells. Moreover, anti-HSV-1 antibodies present in the cerebrospinal fluid of patients with HSV-1 encephalitis efficiently neutralize viral infection, block viral cell-to-cell transmission, and inhibit propagation of pathological Tau. Thus, latent HSV-1 infection of the brain could contribute to neurodegenerative disease progression, and antiviral antibodies may represent a potential therapeutic strategy.

neuroscience↗

Gain efficiency with streamlined and automated data processing: Examples from high-throughput monoclonal antibody production

Data management and sample tracking in complex biological workflows are essential steps to ensure necessary documentation and guarantee the reusability of data and metadata. Currently, these steps pose challenges related to correct annotation and labeling, error detection, and safeguarding the quality of documentation. With growing acquisition of biological data and the expanding automatization of laboratory workflows, manual processing of samples is no longer favorable, as it is time- and resource-consuming, is prone to biases and errors, and lacks scalability and standardization. Thus, managing heterogeneous biological data calls for efficient and tailored systems, especially in laboratories run by biologists with limited computational expertise. Here, we showcase how to meet these challenges with a modular pipeline for data processing, facilitating the complex production of monoclonal antibodies from single B-cells. We present best practices for development of data processing pipelines concerned with extensive acquisition of biological data that undergoes continuous manipulation and analysis. Moreover, we assess the versatility of proposed design principles through a proof-of-concept data processing pipeline for automated induced pluripotent stem cell culture and differentiation. We show that our approach streamlines data management operations, speeds up experimental cycles and leads to enhanced reproducibility. Finally, adhering to the presented guidelines will promote compliance with FAIR principles upon publishing.

cell biology↗

An ex vivo human model for safety assessment of immunotoxicity of engineered nanomaterials

The unique physicochemical properties of nanomaterials (NM) and engineered nanomaterials (ENM) have pushed their use in many applications ranging from medicine to the food industry, textiles, and many more fields. Thus, human exposure to NM and ENM is growing by the day. However, the current toxicity tests do not reflect the special characteristics of ENM and are not developed for ENM risk assessment. Here we propose a high-throughput cell-based assay using human peripheral blood mononuclear cells (PBMCs) that can monitor the effects of NM and ENM on cytotoxicity and innate immunity. The proposed assay is fully automated and miniaturized, with excellent assay performance parameters (Z-score >0.5), amenable for large screening campaigns in industrial setting. Immunotoxicity data for ENM safety assessment are collected in dose-response format. At different states, multiparametric readouts for cytotoxicity, and innate immunity are conducted in a combinatorial method, avoiding ENM-induced bias by endotoxin contamination. Integrating this high-dimensional data, allows (i) holistic safety assessment of immunotoxicity effects caused by ENM, classifying safe and toxic ENM phenotypes, and (ii) to deconvolve mode of action of the ENM effect on the PBMCs. As added value the data obtained can be used to troubleshoot ENM or for a safe-by-design approach in product development.

immunology↗

Evolution of a new testis-specific functional promotor within the highly conserved Map2k7 gene of the mouse

Map2k7 (synonym Mkk7) is a conserved regulatory kinase gene and a central component of the JNK signaling cascade with key functions during cellular differentiation. It shows complex transcription patterns and different transcript isoforms are known in the mouse (Mus musculus). We have previously identified a newly evolved testis specific transcript for the Map2k7 gene in the subspecies M. m. domesticus. Here, we identify the new promotor that drives this transcript and find that its transcript codes for an open reading frame (ORF) of 50 amino acids. The new promotor was gained in the stem lineage of closely related mouse species, but was secondarily lost in the subspecies M. m. musculus and M. m. castaneus. A single mutation can be correlated with its transcriptional activity in M. m. domesticus and cell culture assays demonstrate the capability of this mutation to drive expression. A mouse knock-out line in which the promotor region of the new transcript is deleted reveals a functional contribution of the newly evolved promotor to sperm motility and to the spermatid transcriptome. Our data show that a new functional transcript (and possibly protein) can evolve within an otherwise highly conserved gene, supporting the notion of regulatory changes contributing to the emergence of evolutionary novelties.

evolutionary biology↗

Fog signaling has diverse roles in epithelial morphogenesis in insects

The Drosophila Fog pathway represents one of the best-understood signaling cascades controlling epithelial morphogenesis. During gastrulation, Fog induces apical cell constrictions that drive the invagination of mesoderm and posterior gut primordia. The cellular mechanisms underlying primordia internalization vary greatly among insects and recent work has suggested that Fog signaling is specific to the fast mode of gastrulation found in some flies. On the contrary, here we show in the beetle Tribolium, whose development is broadly representative for insects, that Fog has multiple morphogenetic functions. It modulates mesoderm internalization and controls a massive posterior infolding involved in gut and extraembryonic development. In addition, Fog signaling affects blastoderm cellularization, primordial germ cell positioning and cuboidal-to-squamous cell shape transitions in the extraembryonic serosa. Comparative analyses with two other distantly related insect species reveals that Fogs role during cellularisation is widely conserved and therefore might represent the ancestral function of the pathway.

developmental biology↗