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Biology subjects

Sounart, H.

Publications and source records attributed to Sounart, H..

4 recordsLinked to original sources

ProbeST: a custom probe design pipeline for Spatial Transcriptomics

Probe-based Spatial Transcriptomics profiles spatially-resolved transcriptomes using gene-specific probe pairs for both Formalin-fixed paraffin-embedded (FFPE) and Fresh Frozen samples. However, its applicability is restricted to human and mouse studies, due to commercial probe set availability. Here, we present ProbeST, an open-source computational pipeline for designing custom probe sets for any organism of interest. We validated ProbeST on FFPE mouse enteroids infected with Salmonella enterica serovar Typhimurium, using custom pathogen probes with the available mouse probe panel. We simultaneously detected host and pathogen transcripts, demonstrating high probe specificity and enabling identification of inflammatory response host genes Mefv, Tnf, and Anxa1 colocalizing to pathogen genes. The reproducible ProbeST workflow expands probe-based Spatial Transcriptomics to studies of non-model organisms and host-pathogen interactions.

bioinformatics↗

Miniature spatial transcriptomics for studying parasite-endosymbiont relationships at the micro scale

Several important human infectious diseases are caused by microscale-sized parasitic nematodes like filarial worms. Filarial worms have their own spatial tissue organization and to uncover this tissue structure, we need methods that can spatially resolve these miniature specimens. Most filarial worms evolved a mutualistic association with endosymbiotic bacteria Wolbachia, however, the mechanisms underlying the dependency of filarial worms on the fitness of these bacteria remain unknown. As Wolbachia is essential for the development, reproduction, and survival of filarial worms, we focused on studying a posterior region containing reproductive tissue and developing embryos of adult female Brugia malayi worms. To spatially explore how Wolbachia interacts with the worms reproductive system, we performed a spatial characterization using Spatial Transcriptomics (ST) across our region of interest. We provide a proof-of-concept for miniature-ST to explore spatial gene expression patterns in small sample types, demonstrating the methods ability to uncover nuanced tissue region expression patterns, observe the spatial localization of key B. malayi - Wolbachia pathway genes, and co-localize the B. malayi spatial transcriptome in Wolbachia tissue regions. We envision our approach to open up new scenarios for the study of infectious diseases caused by micro-scale parasitic worms.

genomics↗

Dual spatially resolved transcriptomics for SARS-CoV-2 host-pathogen colocalization studies in humans

To advance our understanding of cellular host-pathogen interactions, technologies that facilitate the co-capture of both host and pathogen spatial transcriptome information are needed. Here, we present an approach to simultaneously capture host and pathogen spatial gene expression information from the same formalin-fixed paraffin embedded (FFPE) tissue section using the spatial transcriptomics technology. We applied the method to COVID-19 patient lung samples and enabled the dual detection of human and SARS-CoV-2 transcriptomes at 55 m resolution. We validated our spatial detection of SARS-CoV-2 and identified an average specificity of 94.92% in comparison to RNAScope and 82.20% in comparison to in situ sequencing (ISS). COVID-19 tissues showed an upregulation of host immune response, such as increased expression of inflammatory cytokines, lymphocyte and fibroblast markers. Our colocalization analysis revealed that SARS-CoV-2+ spots presented shifts in host RNA metabolism, autophagy, NF{kappa}B, and interferon response pathways. Future applications of our approach will enable new insights into host response to pathogen infection through the simultaneous, unbiased detection of two transcriptomes.

genomics↗

Escape from bacterial diversity: potential enemy release in invading yellow starthistle (Centaurea solstitialis) microbiomes

O_LIInvasive species could benefit from introduction to locations with favorable species interactions. Microbiomes are an important source of interactions that vary across regions. We examine whether bacterial communities could explain more favorable microbial interactions in highly invasive populations of yellow starthistle.\nC_LIO_LIWe sequenced amplicons of prokaryotic 16S rRNA genes to characterize bacterial community composition in the phyllosphere, ectorhizosphere, and endorhizosphere of plants from seven invading populations in California, USA and eight native populations in Europe. We tested for differentiation of microbiomes by geography, plant compartment, and plant genotype.\nC_LIO_LIBacterial communities differed significantly between native and invaded ranges within plant compartments, with consistently lower diversity in plants from the invaded range. Genera containing known plant pathogens also showed lower diversity in invaded range plants. The diversity of bacteria in roots was positively correlated with plant genotype diversity within both ranges, but this relationship did not explain microbial differences between ranges.\nC_LIO_LIOur findings reveal changes in the composition and diversity of bacterial interactions in invading plants, consistent with observations of altered soil interactions in this invasion. These results call for further study of the sources of variation in microbiomes and the potential for bacteria to facilitate invasion success.\nC_LI

plant biology↗