Biology subjects

Roth, W.

Publications and source records attributed to Roth, W..

3 recordsLinked to original sources

bioRxiv2024

MICROBIOME-MODIFIED METABOLITES PREDICT ACUTE BRAIN INJURY OUTCOME

BACKGROUNDAcute brain injury (ABI)-mediated disruption of the microbiome may potentiate inflammation and secondary brain injury (SBI). However, microbial-specific mediators and mechanisms remain unclear. METHODSThirty-five consecutive patients with ABI admitted to the neuroscience critical care unit at the University of Chicago were prospectively studied. Injury severity at hospital admission was assessed using the Injury Severity Score (ISS) and the Glasgow Coma Scale (GCS). Final neurologic function was assessed via the Glasgow Outcome Score extended (GOSe). Serum, plasma, and stool targeted metabolomics, as well as stool shotgun metagenomics, were performed on longitudinal samples collected during hospitalization. RESULTSMultivariate analysis identified microbiome-modified metabolites that were positively and negatively associated with functional outcomes after ABI. Novel identification of conjugated bile acid (BA) species and vitamin B12 precursors indicative of outcome were detected in the first collected samples (within 48 hours). Network analysis revealed greater integration of negatively associated metabolites across tissues and identified tauro-/-muricholic acid (TMCA) as central to the cross-tissue metabolomes. CONCLUSIONSMicrobiome metabolites may be useful in assessing brain injury outcomes to inform treatment. Bile acid species transformed by the gut microbiome are predictive of ABI outcome.

neuroscience↗

bioRxiv2023

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↗

bioRxiv2021

Human iPSC-derived brain endothelial microvessels in a multi-well format enable permeability screens of anti-inflammatory drugs

Optimizing drug candidates for blood-brain barrier (BBB) penetration in humans remains one of the key challenges and many devastating brain diseases including neurodegenerative diseases still do not have adequate treatments. So far, it has been difficult to establish state-of-the-art human stem cell derived in vitro models that mimic physiological barrier properties including a 3D microvasculature in a format that is scalable enough to screen drugs for BBB penetration in early drug development phases. To address this challenge, we established human induced pluripotent stem cell (iPSC)-derived brain endothelial microvessels in a standardized and scalable multi-well plate format. iPSC-derived brain microvascular endothelial cells (BMECs) were supplemented with primary cell conditioned media and grew to intact microvessels in 10 days of culturing. Produced microvessels show a typical BBB phenotype including endothelial protein expression, tight-junctions and polarized localization of efflux transporter. Microvessels exhibited physiological relevant trans-endothelial electrical resistance (TEER), were leak-tight for 10 kDa dextran-Alexa 647 and strongly limited the permeability of sodium fluorescein (NaF). Permeability tests with reference compounds confirmed the suitability of our model as platform to identify potential BBB penetrating anti-inflammatory drugs. In summary, the here presented brain microvessel platform recapitulates physiological properties and allows rapid screening of BBB permeable anti-inflammatory compounds that has been suggested as promising substances to cure so far untreatable neurodegenerative diseases.

cell biology↗