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Biology subjects

Charrier, E.

Publications and source records attributed to Charrier, E..

3 recordsLinked to original sources

The development and validation of long-read ITS-1/5.8S/ITS-2 nemabiome metabarcoding using Oxford Nanopore Technologies (ONT) sequencing for ovine gastrointestinal nematodes and the design of primers suitable for its direct application to fecal DNA.

ITS-2 rRNA nemabiome metabarcoding is increasingly used to characterize gastrointestinal nematode (GIN) communities. While powerful, current approaches have some limitations in their flexibility and applicability to smaller-scale studies and diagnostic use. The short read lengths provided by the Illumina platform may lack discriminatory power for some closely related species and also pose a challenge for new marker selection and primer design. To address these challenges, we have developed ITS-1/5.8S/ITS-2 rRNA Oxford Nanopore Technologies (ONT) long-read metabarcoding for ovine gastrointestinal nematodes. Samples from two previous field studies, from UK and western Canadian sheep farms were used. ITS-1/5.8S/ITS-2 long-read metabarcoding showed strong concordance with prior ITS-2 data for the major GIN species in both datasets, with minor discrepancies for some low abundance taxa mainly due to differences in reference sequence database representation. We also used the PrimerTC tool to design a new primer pair, EC1 and EC2, to minimize the amplification of off-target fungal sequences derived from fecal DNA and maximize the nematode sequence read depth when ONT ITS-1/5.8S/ITS-2 metabarcoding was applied directly to ovine fecal stool DNA. In summary, ITS-1/5.8S/ITS-2 ONT long-read nemabiome metabarcoding showed good agreement with ITS-2 metabarcoding and the use of primer pair EC1/EC2 should make the approach more tolerant of fecal contamination of parasite material, with the potential for direct application to ovine fecal DNA. Overall, these new developments should make nemabiome metabarcoding more accessible, discriminating, and flexible for both research and diagnostic applications.

molecular biology↗

IL-33 drives polyfunctionality and antitumor activity of a unique ST2+ NK cell population

Despite increasing recognition of NK cell diversity, their functional heterogeneity and its relevance to cancer remain incompletely understood. Here, we show that IL-12 promotes the emergence of a distinct ST2/IL33R+ NK cell state poised for robust activation by IL-33. Human ST2 NK cells exhibit a unique transcriptional program, an intermediate differentiation state between CD56bright and CD56dim NK cells, and enhanced polyfunctionality combining proliferation, cytokine production, and cytotoxicity. ST2 NK cells were identified across human and murine tumor datasets. In murine breast cancer models, IL-33 and IL-12 cooperated to elicit potent antitumor responses dependent on IFN-{gamma} and ST2 NK cells. In cancer patients, IL-12 unleashed tumor-infiltrating ST2 NK cells' potential to produce IFN-g in response to IL-33, an IL33hi-NKhi score was associated with prolonged survival, and ST2 NK cells correlated with improved immunotherapy response. Collectively, these findings identify ST2 NK cells as a therapeutically actionable NK cell state promoting antitumor immunity.

cancer biology↗

Vimentin intermediate filaments mediate cell shape on visco-elastic substrates

The ability of cells to take and change shape is a fundamental feature underlying development, wound repair, and tissue maintenance. Central to this process is physical and signaling interactions between the three cytoskeletal polymeric networks: F-actin, microtubules, and intermediate filaments (IFs). Vimentin is an IF protein that is essential to the mechanical resilience of cells and regulates cross-talk amongst the cytoskeleton, but its role in how cells sense and respond to the surrounding extracellular matrix is largely unclear. To investigate vimentins role in substrate sensing, we designed polyacrylamide hydrogels that mimic the elastic and viscoelastic nature of in vivo tissues. Using wild-type and vimentin-null mouse embryonic fibroblasts, we show that vimentin enhances cell spreading on viscoelastic substrates, even though it has little effect in the limit of purely elastic substrates. Our results provide compelling evidence that the vimentin cytoskeletal network is a physical modulator of how cells sense and respond to mechanical properties of their extracellular environment.

biophysics↗