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Biology subjects

Reynolds, T.

Publications and source records attributed to Reynolds, T..

5 recordsLinked to original sources

Scaling up spatial transcriptomics for large-sized tissues: uncovering cellular-level tissue architecture beyond conventional platforms with iSCALE

Recent advances in spatial transcriptomics (ST) technologies have transformed our ability to profile gene expression while retaining the crucial spatial context within tissues. However, existing ST platforms suffer from high costs, long turnaround times, low resolution, limited gene coverage, and small tissue capture areas, which hinder their broad applications. Here we present iSCALE, a method that predicts super-resolution gene expression and automatically annotates cellular-level tissue architecture for large-sized tissues that exceed the capture areas of standard ST platforms. The accuracy of iSCALE were validated by comprehensive evaluations, involving benchmarking experiments, immunohistochemistry staining, and manual annotation by pathologists. When applied to multiple sclerosis human brain samples, iSCALE uncovered lesion associated cellular characteristics that were undetectable by conventional ST experiments. Our results demonstrate iSCALEs utility in analyzing large-sized tissues with automatic and unbiased tissue annotation, inferring cell type composition, and pinpointing regions of interest for features not discernible through human visual assessment.

genomics↗

A Time Machine for Taxonomy

The NCBI Taxonomy Database is the primary resource for linking genomic information to taxonomic relationships, widely used across scientific disciplines and critically important to bioinformatics. This database is continuously changing as researchers discover and refine taxonomic relationships. Yet, tracking and comparing past taxonomic states is challenging due to frequent changes and the need to sift through numerous historical snapshots. To address this, we developed the Taxonomy Time Machine: a database for storing many snapshots of a taxonomic tree in a space-efficient manner. We have also created a web-based and programmatic (API) interface to make this data more accessible. This tool is capable of accurately reconstructing taxonomic lineages at any point in the history of the NCBI Taxonomy Database. We demonstrate that this tool is both perfectly accurate and significantly more efficient than loading and querying individual taxonomy snapshots, enabling its use on desktop computers as well as commodity web servers. We have made this tool available on the web (https://taxonomy.onecodex.com) as well as open source under the MIT license (https://github.com/onecodex/taxonomy-time-machine).

bioinformatics↗

'Off-pore' nucleoporins relocalize heterochromatic breaks through phase separation

Phase separation forms membraneless compartments in the nuclei, including by establishing heterochromatin "domains" and repair foci. Pericentromeric heterochromatin mostly comprises repeated sequences prone to aberrant recombination, and "safe" homologous recombination (HR) repair of these sequences requires the movement of repair sites to the nuclear periphery before Rad51 recruitment and strand invasion. How this mobilization initiates is unknown, and the contribution of phase separation to these dynamics is unclear. Here, we show that Nup98 nucleoporin is recruited to heterochromatic repair sites before relocalization through Sec13 or Nup88 nucleoporins, and downstream from the Smc5/6 complex and SUMOylation. Remarkably, the phase separation properties of Nup98 are required and sufficient to mobilize repair sites and exclude Rad51, thus preventing aberrant recombination while promoting HR repair. Disrupting this pathway results in heterochromatin repair defects and widespread chromosome rearrangements, revealing a novel "off-pore" role for nucleoporins and phase separation in nuclear dynamics and genome integrity in a multicellular eukaryote. HighlightsO_LINup88 and Sec13 recruit Nup98 to heterochromatic DSBs downstream from Smc5/6 C_LIO_LINup88, Sec13 and Nup98 promote repair focus mobilization in heterochromatin off-pore C_LIO_LINup98 excludes Rad51 from repair sites inside the heterochromatin domain C_LIO_LIPhase separation by Nup98 is required and sufficient for relocalization and Rad51 exclusion C_LI

cell biology↗

Genome-scale analysis of interactions between genetic perturbations and natural variation

Interactions between genetic perturbations and segregating loci can cause perturbations to show different phenotypic effects across genetically distinct individuals. To study these interactions on a genome scale in many individuals, we used combinatorial DNA barcode sequencing to measure the fitness effects of 7,700 CRISPRi perturbations targeting 1,712 distinct genes in 169 yeast cross progeny (or segregants). We identified 460 genes whose perturbation has different effects across segregants. Several factors caused perturbations to show variable effects, including baseline segregant fitness, the mean effect of a perturbation across segregants, and interacting loci. We mapped 234 interacting loci and found four hub loci that interact with many different perturbations. Perturbations that interact with a given hub exhibit similar epistatic relationships with the hub and show enrichment for cellular processes that may mediate these interactions. These results suggest that an individuals response to perturbations is shaped by a network of perturbation-locus interactions that cannot be measured by approaches that examine perturbations or natural variation alone.

genetics↗

Finding human gene-disease associations using a Network Enhanced Similarity Search (NESS) of multi-species heterogeneous functional genomics data

Disease diagnosis and treatment is challenging in part due to the misalignment of diagnostic categories with the underlying biology of disease. The evaluation of large-scale genomic experimental datasets is a compelling approach to refining the classification of biological concepts, such as disease. Well-established approaches, some of which rely on information theory or network analysis, quantitatively assess relationships among biological entities using gene annotations, structured vocabularies, and curated data sources. However, the gene annotations used in these evaluations are often sparse, potentially biased due to uneven study and representation in the literature, and constrained to the single species from which they were derived. In order to overcome these deficiencies inherent in the structure and sparsity of these annotated datasets, we developed a novel Network Enhanced Similarity Search (NESS) tool which takes advantage of multi-species networks of heterogeneous data to bridge sparsely populated datasets. NESS employs a random walk with restart algorithm across harmonized multi-species data, effectively compensating for sparsely populated and noisy genomic studies. We further demonstrate that it is highly resistant to spurious or sparse datasets and generates significantly better recapitulation of ground truth biological pathways than other similarity metrics alone. Furthermore, since NESS has been deployed as an embedded tool in the GeneWeaver environment, it can rapidly take advantage of curated multi-species networks to provide informative assertions of relatedness of any pair of biological entities or concepts, e.g., gene-gene, gene-disease, or phenotype-disease associations. NESS ultimately enables multi-species analysis applications to leverage model organism data to overcome the challenge of data sparsity in the study of human disease. Availability and ImplementationImplementation available at https://geneweaver.org/ness. Source code freely available at https://github.com/treynr/ness. Author summaryFinding consensus among large-scale genomic datasets is an ongoing challenge in the biomedical sciences. Harmonizing and analyzing such data is important because it allows researchers to mitigate the idiosyncrasies of experimental systems, alleviate study biases, and augment sparse datasets. Additionally, it allows researchers to utilize animal model studies and cross-species experiments to better understand biological function in health and disease. Here we provide a tool for integrating and analyzing heterogeneous functional genomics data using a graph-based model. We show how this type of analysis can be used to identify similar relationships among biological entities such as genes, processes, and disease through shared genomic associations. Our results indicate this approach is effective at reducing biases caused by sparse and noisy datasets. We show how this type of analysis can be used to aid the classification gene function and prioritization of genes involved in substance use disorders. In addition, our analysis reveals genes and biological pathways with shared association to multiple, co-occurring substance use disorders.

bioinformatics↗