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Biology subjects

Evans, B.

Publications and source records attributed to Evans, B..

At least 19 recordsLinked to original sources

A foundational model for joint sequence-function multi-species modeling at scale for long-range genomic prediction

Genomic prediction and design require models that integrate local sequence features with long-range regulatory dependencies spanning hundreds of kilobases to megabases. Existing approaches have made substantial progress along complementary axes: supervised sequence-to-function models achieve high accuracy for specific assays and organisms, self-supervised genomic foundation models learn transferable representations from large-scale sequence data, and conditional generative models enable principled sequence design guided by functional objectives. However, these strengths are typically realized in isolation--across distinct model classes, architectures, and training regimes--limiting the ability to combine long-context, base-resolution prediction, functional modeling, and controllable generation within a single efficient framework that generalizes across organisms and modalities. Here we introduce Nucleotide Transformer v3 (NTv3), a multi-species foundation model that unifies representation learning, functional-track and genome-annotation prediction, and controllable sequence generation within a common backbone. NTv3 uses a U-Net-like architecture to enable single-base tokenization and efficient modeling of contexts up to 1 Mb. We pre-train NTv3 on 9 trillion base pairs from OpenGenome2 using base-resolution masked language modeling, followed by post-training with a joint objective that integrates continued self-supervision with supervised learning on [~]16,000 functional tracks and annotation labels from 24 animal and plant species. After post-training, NTv3 achieves state-of-the-art accuracy for functional-track prediction and genome annotation across species, outperforming leading sequence-to-function and foundation-model baselines on established benchmarks and on the new NO_SCPLOWTVC_SCPLOW 3 BO_SCPLOWENCHMARKC_SCPLOW, a controlled downstream fine-tuning suite in a standardized 32 kb input / base-resolution output setting. We further show that NTv3 consolidates a shared regulatory grammar across tasks, enabling coherent long-range genome-to-function inference and variant-associated remodeling. Finally, we fine-tune NTv3 into a controllable generative model via masked diffusion language modeling and use it to design enhancer sequences with specified activity levels and promoter selectivity. We validate these designs experimentally, showing that generated enhancers recapitulate the intended activity stratification and achieve the desired promoter-specific activation in cellulo. We release the NTv3 model family together with code and practical cookbooks for long-context training, multispecies post-training, fine-tuning, interpretation, and sequence design.

genomics↗

Locomotion Selectively Amplifies Sensitizing Neurons in Primary Visual Cortex

Sensory processing in the cortex reflects both adaptation to external stimuli and changes in internal state. To investigate how these processes interact in layer 2/3 of mouse V1 we combined calcium imaging, optogenetics and circuit modelling. We find that locomotion preferentially increases the responses of pyramidal cells (PCs) that sensitize during visual stimulation compared to those that depress. A model explains this differential modulation through: (i) variations in the strength of PV and SST connectivity to individual PCs, (ii) broad locomotion-dependent weakening of PC and PV synapses, and (iii) reduced SST inhibition targeting sensitizing PCs. Differences in PV:SST input ratios across sensitizing and depressing PCs can be reproduced by a random-connectivity model based on measured interneuron densities, connection probabilities and synaptic strengths. Thus, stochastic variation in local inhibitory connectivity can explain much of the functional heterogeneity across PCs, while state-dependent modulation of inhibitory synapses reorganizes this balance during locomotion to bias cortical population activity towards sensitizing dynamics. The apparently paradoxical combination of increased PC response but decreased synaptic strength is consistent with a state-dependent gating mechanism that boosts signals leaving V1 while simultaneously preventing disruption of the local excitatory-inhibitory balance required for stable computation.

neuroscience↗

A spatial long-read approach at near-single-cell resolution reveals developmental regulation of splicing and polyadenylation sites in distinct cortical layers and cell types.

Genome-wide single-cell and spatial long-read approaches have gained traction, but mostly lack single-cell resolution - and yield limited read lengths. Here, we introduce spatial ISOform sequencing (Spl-ISO-Seq), which reveals exons and polyadenylation sites from long reads with near-single-cell resolution. Spl-ISO-Seq selects long cDNAs and doubles to triples read lengths compared to standard preparations. Adding a highly specific software tool (Spl-ISOquant) and comparing human post-mortem pre-puberty samples of the visual cortex (8-11 years) to post-puberty samples (16-19 years), we find that cortical layers harbor stronger splicing and poly(A)-site regulation than the adjacent white matter, with enrichment of multiple protein-domain types. For oligodendrocytes however, developmental splicing changes are stronger in white matter. Among cortical layers, layer 4 has the most developmental changes in alternative-exon inclusion in excitatory neurons and in poly(A) sites. We also find many repeat elements, especially ERV1 long terminal repeats downstream of developmentally-regulated layer 4 exons. Overall, alternative splicing changes are linked to synapses - specifically at the post-synapse. Age-linked splicing changes in layers 1-3 and 4 are associated with autism spectrum disorder but not with schizophrenia, amyotrophic lateral sclerosis and Alzheimers disease. These results root developmental splicing changes during puberty and the resulting protein changes in specific layers and cell types. More generally, our new technologies enable new observations for any complex tissue.

genomics↗

Genome-Wide Associations within Diverse Wild Apple Germplasm for Postharvest Blue Mold Resistance to Penicillium expansum

Post-harvest disease caused by the blue mold fungus, Penicillium expansum, accounts for a substantial proportion of economic losses in United States apple industry. Multiple modes of entry in the apple supply chain, plus emerging fungicide resistance, limit the current and long-term viability of using chemical controls alone. Previous phenotypic screens of Malus accessions in the USDA-ARS apple germplasm have identified varying levels of blue mold disease resistance in some wild apple accessions and hybrids. These wild apple species contain reservoirs of genetic resistance that can be integrated into apple breeding programs to complement the previously identified qM-Pe3.1 marker from M. sieversii. We sought to identify these novel loci by combining historical phenotypes of the USDA-ARS wild apple germplasm with low-pass genomic sequencing to perform association mapping. Multi-locus mixed models identified five single nucleotide polymorphisms (SNPs) significantly associated with reduction of post-harvest rot under high concentration of P. expansum inoculum, and one SNP associated under low inoculum concentration. Within a 25,000 base pair window of these SNPs, we found candidate genes encoding proteins with known pathogen immune response and defense roles, such as a Cobra-like 7, flavin monooxygenase, LRR receptors, PR5-like receptor kinase, and a putative resistance protein RGA3. We present these loci as targets for identifying accessions with beneficial alleles that can be targeted for fine mapping and use in Malus breeding programs to achieve M. domestica lines with natural post-harvest rot resistance.

genetics↗

Random connectivity generates inhibitory microcircuits that decorrelate adaptation in visual cortex

Inhibitory neurons are fundamental to sensory processing in the cortex but the rules governing their connections with excitatory neurons are unclear. Are pyramidal cells with different functions generated through specific or random connections? We used two-photon imaging, optogenetics and modelling to investigate opposing forms of adaptation in layer 2/3 of mouse visual cortex. We find that a slow modulatory drive acts differentially depending on the relative strength of inputs that individual pyramidal cells receive from parvalbumin-positive and somatostatin-positive interneurons. The number of depressing and sensitizing pyramidal cells could be explained quantitatively by the simplest connectivity rule - all inhbitory synapses made randomly. The functional heterogeneity of the pyramidal cell population therefore begins with general statistics of the connectome - interneurons are much sparser and connect with probabilities far less than one. The resulting "patchwork" of inhibitory microcircuits causes modulatory inputs to strongly decorrelate pyramidal cells on the behavioural time-scale of seconds.

neuroscience↗

Multiplex PCR reveals population structure in an inbred communal bird.

We sampled communally breeding p[u]keko (Porphyrio melanotus melanotus, family Rallidae) populations on the North (T[a]wharanui Park) and South (Otokia Reserve) Islands of New Zealand that differ in climate and ecology. North Island populations have year-round territories, philopatry, and form kin groups, resulting in inbreeding. South Island populations have seasonal territories, high dispersal rates, and form non-kin groups, leading to outbreeding. Given behavioural evidence of inbreeding we predicted that the North Island population would exhibit lower heterozygosity and higher inbreeding coefficients than the South Island population. We hypothesized that the South Island population originated via a range expansion from the north and predicted that South Island birds would have lower allelic diversity due to founder effects. To test these predictions, we developed microsatellite primers, optimized multiplex PCRs, and genotyped breeding groups from the North and South Island. Breeding groups from North Island were genetically differentiated, whereas population structure was not detected in the South Island birds. North Island birds had higher inbreeding coefficients and levels of within group kinship, but not allelic diversity, compared to South Island birds. Our results are thus inconclusive about whether the South Island population originated via a range expansion from the north. This pilot study validated microsatellite markers and PCR methods and is the first genetic analysis of population structure and relatedness within communal breeding p[u]keko groups. These genetic tools will be used for larger-scale studies to help resolve the origins of the South Island population and provide further insights into the effects of ecology and behaviour on inbreeding, reproductive success, and population demographics in this species. P[u]keko may provide an excellent model for experimental analyses of inbreeding effects on wild avian populations without the attendant concerns that come with small, endangered populations. This work may thus inform conservation efforts, including translocations of endangered species.

genetics↗

The unknown lipids project: harmonized methods improve compound identification and data reproducibility in an inter-laboratory untargeted lipidomics study

Untargeted lipidomics allows analysis of a broader range of lipids than targeted methods and permits discovery of unknown compounds. Previous ring trials have evaluated the reproducibility of targeted lipidomics methods, but inter-laboratory comparison of compound identification and unknown feature detection in untargeted lipidomics has not been attempted. To address this gap, five laboratories analyzed a set of mammalian tissue and biofluid reference samples using both their own untargeted lipidomics procedures and a common chromatographic and data analysis method. While both methods yielded informative data, the common method improved chromatographic reproducibility and resulted in detection of more shared features between labs. Spectral search against the LipidBlast in silico library enabled identification of over 2,000 unique lipids. Further examination of LC-MS/MS and ion mobility data, aided by hybrid search and spectral networking analysis, revealed spectral and chromatographic patterns useful for classification of unknown features, a subset of which were highly reproducible between labs. Overall, our method offers enhanced compound identification performance compared to targeted lipidomics, demonstrates the potential of harmonized methods to improve inter-site reproducibility for quantitation and feature alignment, and can serve as a reference to aid future annotation of untargeted lipidomics data.

systems biology↗

COR27/28 Regulate the Evening Transcriptional Activity of the RVE8 and LNK1/2 Circadian Complex

The timing of many molecular and physiological processes in plants occurs at a specific time of day. These daily rhythms are driven by the circadian clock, a master timekeeper that uses daylength and temperature to maintain rhythms of approximately 24 hours in various clock-regulated phenotypes. The circadian MYB-like transcription factor REVEILLE 8 (RVE8) interacts with its transcriptional coactivators NIGHT LIGHT INDUCIBLE AND CLOCK REGULATED 1 (LNK1) and LNK2 to promote the expression of evening-phased clock genes and cold tolerance factors. While genetic approaches have commonly been used to discover new connections within the clock and between other pathways, here we use affinity purification coupled with mass spectrometry to discover time-of-day-specific protein interactors of the RVE8-LNK1/2 complex. Among the interactors of RVE8/LNK1/LNK2 were COLD REGULATED GENE 27 (COR27) and COR28, which were coprecipitated in an evening-specific manner. In addition to COR27/28, we found an enrichment of temperature-related interactors that led us to establish a novel role for LNK1/2 in temperature entrainment of the clock. We established that RVE8, LNK1, and either COR27 or COR28 form a tripartite complex in yeast and that the effect of this interaction in planta serves to antagonize transcriptional activation of RVE8 target genes through mediating RVE8 protein degradation in the evening. Together, these results illustrate how a proteomic approach identified time-of-day-specific protein interactions and a novel RVE8-LNK-COR protein complex that implicates a new regulatory mechanism for circadian and temperature signaling pathways.

plant biology↗

Acinetobacter baumannii sampled from cattle and pigs represent novel clones

Acinetobacter baumannii is a very important human pathogen. Nonetheless, we know very little about non-human isolates of A. baumannii. Here we determine the genomic identity of 15 cattle and pig isolates, as well their antibiotic and virulence genetic determinants, and compare them to the main human clinical international clones.

microbiology↗

Characterizing dry mass and volume changes in human multiple myeloma cells upon treatment with proteotoxic and genotoxic drugs

Multiple myeloma (MM) is a cancer of terminally differentiated plasma cells. MM remains incurable, but overall survival of patients has progressively increased over the past two decades largely due to novel agents such as proteasome inhibitors (PI) and the immunomodulatory agents. While these therapies are highly effective, MM patients can be de novo resistant and acquired resistance with prolonged treatment is inevitable. There is growing interest in early, accurate identification of responsive versus non-responsive patients, however limited sample availability and need for rapid assays are limiting factors. Here, we test dry mass and volume as label-free biomarkers to monitor early response of MM cells to treatment with bortezomib, doxorubicin, and ultraviolet light. For the dry mass measurement, we use two types of phase-sensitive optical microscopy techniques: digital holographic tomography and computationally-enhanced quantitative phase microscopy. We show that human MM cell lines (RPMI8226, MM.1S, KMS20, and AMO1) increase dry mass upon bortezomib treatment. This dry mass increase after bortezomib treatment occurs as early as 1 hour for sensitive cells and 4 hours for all tested cells. We further confirm this observation using primary multiple myeloma cells derived from patients and show that a correlation exists between increase in dry mass and sensitivity to bortezomib, supporting the use of dry mass as a biomarker. The volume measurement using Coulter counter shows a more complex behavior; RPMI8226 cells increase the volume at an early stage of apoptosis, but MM.1S cells show the volume decrease typically observed with apoptotic cells. Altogether, this cell study presents complex kinetics of dry mass and volume at an early stage of apoptosis, which may serve as a basis for the detection and treatment of MM cells.

systems biology↗

A novel paradigm for optimal mass feature peak picking in large scale LC-MS datasets using the isopair: isoLock, autoCredential and anovAlign

Untargeted metabolomics enables direct quantification of metabolites without apriori knowledge of their identity. Liquid chromatography mass spectrometry (LC-MS), a popular method to implement untargeted metabolomics, identifies metabolites via combined mass/charge (m/z) and retention time as mass features. Improvements in the sensitivity of mass spectrometers has increased the complexity of data produced, leading to computational obstacles. One outstanding challenge is calling metabolite mass feature peaks rapidly and accurately in large LC-MS datasets (dozens to thousands of samples) in the presence of measurement and other noise. While existing algorithms are useful, they have limitations that become pronounced at scale and lead to false positive metabolite predictions as well as signal dropouts. To overcome some of these shortcomings, biochemists have developed hybrid computational and carbon labeling techniques, such as credentialing. Credentialing can validate metabolite signals, but is laborious and its applicability is limited. We have developed a suite of three computational tools to overcome the challenges of unreliable algorithms and inefficient validation protocols: isolock, autoCredential and anovAlign. Isolock uses isopairs, or metabolite-istopologue pairs, to calculate and correct for mass drift noise across LC-MS runs. autoCredential leverages statistical features of LC-MS data to amplify naturally present 13C isotopologues and validate metabolites through isopairs. This obviates the need to artificially introduce carbon labeling. anovAlign, an anova-derived algorithm, is used to align retention time windows across samples to accurately delineate retention time windows for mass features. Using a large published clinical dataset as well as a plant dataset with biological replicates across time, genotype and treatment, we demonstrate that this suite of tools is more sensitive and reproducible than both an open source metabolomics pipelines, XCMS, and the commercial software progenesis QI. This software suite opens a new era for enhanced accuracy and increased throughput for untargeted metabolomics.

bioinformatics↗

Restoration of a pond: monitoring water quality and macroinvertebrate community succession

To determine the success of restoration programmes, knowledge of the temporal dynamics in community structure and processes is vital. The water quality and macroinvertebrate community structures of a newly created and an established pond within the same ecosystem were sampled bi-monthly over a year to monitor the development of the new pond. The water quality measures of the ponds were significantly different. Conductivity, salinity, and total dissolved solids levels were also different between the ponds. The colonisation of a macroinvertebrate community in the new pond was rapid, resulting in a 75% resemblance to the established pond by the end of the first year. The pond was colonised by non-insect taxa like Crustacea and Gastropod and then by insects. There was a significant difference in the macroinvertebrate communities of the ponds due to temporal taxonomic composition differences. The high abundance of Diplacodes spp. (perchers), Physa spp. (left-handed pond snail), and Ostracod (seed shrimp) in the new pond contributed to the difference in the community between the two ponds. Cladocera (water fleas) dominated the macroinvertebrate community, and the highest abundance was recorded in August for both ponds. Our results suggest that a newly created pond can have a comparable macroinvertebrate community to nearby established ponds within a year.

ecology↗

Systems-wide Analysis Revealed Shared and Unique Responses to Moderate and Acute High Temperatures in the Green Alga Chlamydomonas reinhardtii

Different intensities of high temperatures affect the growth of photosynthetic cells in nature. To elucidate the underlying mechanisms, we cultivated the unicellular green alga Chlamydomonas reinhardtii under highly controlled photobioreactor conditions and revealed systems-wide shared and unique responses to 24-hour moderate (35{degrees}C) and acute (40{degrees}C) high temperatures and subsequent recovery at 25{degrees}C. We identified previously overlooked unique elements in response to moderate high temperature. Heat at 35{degrees}C transiently arrested the cell cycle followed by partial synchronization, up-regulated transcripts/proteins involved in gluconeogenesis/glyoxylate-cycle for carbon uptake, promoted growth, and increased starch accumulation. Heat at 40{degrees}C arrested the cell cycle, inhibited growth, resulting in carbon uptake over usage and increased starch accumulation. Both high temperatures induced photoprotection, while 40{degrees}C decreased photosynthetic efficiency, distorted thylakoid/pyrenoid ultrastructure, and affected the carbon concentrating mechanism. We demonstrated increased transcript/protein correlation during both heat treatments, suggesting reduced post-transcriptional regulation during heat may help coordinate heat tolerance activities efficiently. During recovery after both heat treatments, transcripts/proteins related to DNA synthesis increased while those involved in photosynthetic light reactions decreased. We propose down-regulating photosynthetic light reactions during DNA replication benefits cell cycle resumption by reducing ROS production. Our results provide potential targets to increase thermotolerance in algae and crops.

cell biology↗

Grb10a knockdown in Danio rerio during early life alters growth and cardiometabolic function associated with a remodelled transcriptome

Embryonic growth trajectory is a risk factor for chronic metabolic and cardiovascular disorder. Grb10 is a negative regulator of the main pathways driving embryonic growth. This study investigates the long-term cardiometabolic consequences and transcriptomic profiles of transient disruption of grb10a expression in Danio rerio. Knockdown was associated with increased embryonic growth (+7%) and metabolic rate (+25%), and decreased heart rate (- 50%) in early life. Juvenile growth and respiratory rate were also elevated (+30% and 7-fold increase respectively). The transcriptome was permanently remodelled by this transient disruption, with dysregulation of multiple growth, cardiac, and metabolic pathways. Phenotypic alteration persisted into adulthood, resulting in a leaner body with elevated skeletal and cardiac muscle content and aerobic scope (43%). This study not only confirms for the first time that transient disruption of a single gene can result in permanent transcriptomic remodelling but correlates this remodelling with persistent alterations to the adult cardiometabolic phenotype.

developmental biology↗

Rooting the animal tree of life

There has been considerable debate about the placement of the root in the animal tree of life, which has emerged as one of the most challenging problems in animal phylogenetics. This debate has major implications for our understanding of the earliest events in animal evolution, including the origin of the nervous system. Some phylogenetic analyses support a root that places the first split in the phylogeny of living animals between sponges and all other animals (the Porifera-sister hypothesis), and others find support for a split between comb jellies and all other animals (Ctenophora-sister). These analyses differ in many respects, including in the genes considered, species considered, molecular evolution models, and software. Here we systematically explore the rooting of the animal tree of life under consistent conditions by synthesizing data and results from 15 previous phylogenomic studies and performing a comprehensive set of new standardized analyses. It has previously been suggested that site-heterogeneous models favor Porifera-sister, but we find that this is not the case. Rather, Porifera-sister is only obtained under a narrow set of conditions when the number of site-heterogeneous categories is unconstrained and range into the hundreds. Site-heterogenous models with a fixed number of dozens of categories support Ctenophora-sister, and cross-validation indicates that such models fit the data just as well as the unconstrained models. Our analyses shed light on an important source of variation between phylogenomic studies of the animal root. The datasets and analyses consolidated here will also be a useful test-platform for the development of phylogenomic methods for this and other difficult problems.

evolutionary biology↗

Integrated transcriptome, DNA methylome and chromatin state accessibility landscapes reveal regulators of Atlantic salmon maturation

Despite sexual development being ubiquitous to vertebrates, the epigenetic mechanisms controlling this fundamental transition remain largely undocumented in many organisms. Through whole-methylome, whole-transcriptome and chromatin landscape sequencing, we discovered global control mechanisms as well as specific regulators of sexual maturation in Atlantic salmon. This large integrated study was based on an experimental time course that successfully sampled the period when Atlantic salmon commence their trajectory towards sexual maturation. Co-analysis of DNA methylome and gene expression changes revealed chromatin remodelling genes arid1b and smarca2 were both significantly hypermethylated and upregulated in the ovary during the onset of maturation. We also observed changes in chromatin state landscape occurred early in the transition and were strongly correlated with fundamental remodelling of gene expression. Finally, we integrated our multiomics datasets to identify trim25 and znf423 as key regulators in the pituitary that underwent 60 fold change in connectivity during the transition to sexual maturation. The study provides a comprehensive view of the spatiotemporal changes involved in a complex trait and opens the door to future efforts aiming to manipulate puberty in an economically important aquaculture species.

genomics↗

Interaction of RSC chromatin remodelling complex with nucleosomes is modulated by H3 K14 acetylation and H2B SUMOylation in vivo

Chromatin remodelling complexes are multi-subunit nucleosome translocases that reorganize chromatin in the context of DNA replication, repair and transcription. A key question is how these complexes find their target sites on chromatin. Here, we use genetically encoded photo-crosslinker amino acids to map the footprint of Sth1, the catalytic subunit of the RSC (remodels the structure of chromatin) complex, on the nucleosome in living yeast. We find that the interaction of the Sth1 bromodomain with the H3 tail depends on K14 acetylation by Gcn5. This modification does not recruit RSC to chromatin but mediates its interaction with neighbouring nucleosomes. We observe a preference of RSC for H2B SUMOylated nucleosomes in vivo and show that this modification moderately enhances RSC binding to nucleosomes in vitro. Furthermore, RSC is not ejected from chromatin in mitosis, but its mode of nucleosome binding differs between interphase and mitosis. In sum, our in vivo analyses show that RSC recruitment to specific chromatin targets involves multiple histone modifications most likely in combination with other components such as histone variants and transcription factors. Key PointsO_LIIn vivo photo-crosslinking reveals the footprint of the ATPase subunit of RSC on the nucleosome. C_LIO_LIRSC binds to H3 K14ac nucleosomes via the C-terminal bromodomain of its ATPase-subunit Sth1. C_LIO_LIRSC preferentially localizes to H2B-SUMOylated nucleosomes. C_LI

biochemistry↗

Quantification and Discovery of Acyl-ACPs by LC-MS/MS

Acyl carrier proteins (ACPs) are the scaffolds for fatty acid biosynthesis in living systems, rendering them essential to a comprehensive understanding of lipid metabolism; however, accurate quantitative methods to assess individual acyl-ACPs do not exist. A robust method was developed to quantify acyl-ACPs at picogram levels. Acyl-ACP elongation intermediates (3-hydroxyacyl-ACPs and 2, 3-trans-enoyl-ACPs), and unexpected medium chain (C10:1, C14:1) and polyunsaturated long chain acyl-ACPs (C16:3) were also identified, indicating the sensitivity of the method and that descriptions of lipid metabolism and ACP function are incomplete. Such ACPs are likely important to medium chain lipid production for fuels and highlight poorly understood lipid remodeling events in the chloroplast. The approach is broadly applicable to Type II FAS systems found in plants, bacteria, and mitochondria of animal and fungal systems because it uses a strategy that capitalizes on a highly conserved Asp-Ser-Leu-Asp (DSLD) amino acid sequence in ACPs to which acyl groups are attached. This allows for sensitive quantification using LC-MS/MS with de novo generated standards and an isotopic dilution strategy and will fill a gap in understanding, providing insights through quantitative exploration of fatty acid biosynthesis processes for optimal biofuels, renewable feed stocks, and medical studies in health and disease.

biochemistry↗