bioRxiv Science⌕ Search

Biology subjects

Robbins, J.

Publications and source records attributed to Robbins, J..

16 recordsLinked to original sources

Structure-free, site-resolved contrastive learningextends small-molecule discovery beyond the reachof structure-based modeling

Virtual screening asks which molecules, among an enormous space of drug-like chemistry, are worth synthesizing and testing against a protein target. Most modern methods answer by building and scoring an explicit three-dimensional pose through molecular docking, or the co-folding models that now approach experimental accuracy. Building these poses presumes a well-defined pocket, and the non-orthosteric, cryptic, and intrinsically disordered sites where unexplored ligandability lies offer none. There, these methods fail to generalize. Here we present Ptarmigan-1, a contrastive model that co-embeds the residues of a protein with candidate small molecules in a shared latent space, from sequence and two-dimensional chemistry alone, and without ever constructing a pose. Engagement reduces to the proximity of precomputed embeddings. Freed from the pose, Ptarmigan-1 trains directly on chemoproteomic and bioactivity data of mixed resolution, scores a compound in ten milliseconds rather than the tens of seconds a co-folding model demands, and resolves each prediction to the residues a compound engages. On well-folded, orthosteric targets it performs comparably to a collection of co-folding and docking models, and on covalent, cryptic, and disordered sites it matches or exceeds them. It localizes reversible and covalent inhibitors to the pockets they engage, even for targets withheld from training, and screens the entire human proteome against a library of 3.4 billion compounds in under a day. By decoupling molecular recognition from structure, Ptarmigan-1 recasts virtual screening as a reusable index that continuously improves as data accumulate.

molecular biology↗

CpG island density predicts CBP/p300 dependency across 3D chromatin clusters

RNA Polymerase II (Pol2) organizes transcription through higher-ordered chromatin clusters that integrate promoter and enhancer interactions to coordinate gene expression. Nevertheless, the features that distinguish unique classes of Pol2-mediated clusters remain to be defined. Here, we identify two distinct classes of Pol2-mediated clusters: one enriched for CpG islands, promoter-promoter interactions, and housekeeping gene expression, and another characterized by high CBP/p300 occupancy, enhancer-promoter looping, and lineage-defining (LD) transcriptional programs. Acute inhibition of CBP/p300 catalytic activity leads to rapid loss of acetylation at enhancers and preferential downregulation of LD genes, resulting in impaired cellular proliferation and activation of apoptotic programs. Integrative machine learning modeling reveals that cluster strength, RNA half-life, and CpG island content as strong predictors of genes sensitive to CBP/p300 inhibition. Together, these findings clarify enhancer-addiction and vulnerability to CBP/p300 inhibition.

cancer biology↗

OCcAM: A tool for rapidly assessing impacts of offshore wind farms on seabirds

Offshore wind energy generation has a key role in reducing reliance on fossil fuels and mitigating climate change. However, it also has the potential to negatively impact the environment, including seabird populations. Quantifying and minimising impacts of wind projects on seabirds is necessary for ensuring compliance with environmental legislation and safeguarding populations. However, the timescales and complexity involved in assessment processes can limit the extent to which estimates can be leveraged towards balancing environmental impact and energy generation. OCcAM is a simple, user-friendly and transparent industry-developed tool allowing rapid assessment of impacts on seabirds from offshore wind developments. Mortality rates of five seabird species are estimated using two UK industry-standard models: the Band collision risk model which predicts fatal collisions with turbine blades, and the matrix-based displacement approach that predicts mortalities associated with distributional responses to the presence of a wind farm. The tool supports simultaneous analysis of up to three input parameter sets and predictions can be expressed as a percentage of a focal population. OCcAM allows auditable analyses to be run quickly and easily with no requirement for specific technical expertise, making it accessible to all stakeholders. It presents a variety of opportunities to facilitate ornithological assessment at strategic to project-specific scales. Here we demonstrate three potential applications of OCcAM: (1) predicting the cumulative impact of Scottish offshore wind farms upon protected seabird populations, (2) updating fatality estimates calculated for worst-case consented values using as-built input parameters and (3) optioneering of project design parameters relative to seabird risk.

ecology↗

Native, Spatiotemporal Profiling of the Global Human Regulome

The regulome, comprising transcription factors, cofactors, chromatin remodelers, and other regulatory proteins, forms the core machinery by which cells interpret signals and execute gene expression programs. Despite its central role in development, disease, and drug response, the regulome remains largely uncharted at scale due to its dynamic, low-abundance, and chromatin-associated nature. Here, we present a method for scalable, regulome profiling for global, compartment-resolved quantification of native regulome proteins. By enriching DNA- and chromatin-associated proteins and profiling them using high-throughput, label-free DIA mass spectrometry, regulome profiling captures chromatin-associated proteins across 36 human cell lines and thousands of perturbations. The resulting Regulome Atlas recovers nearly 60% of known human transcription factors, reveals lineage-specific TF localization, and distinguishes active nuclear engagement from latent, unbound states. We demonstrate that regulome profiles resolve acute immune pathway activation prior to transcriptional changes, identify previously unrecognized drug-induced regulome responses, and enable proteome-scale readouts of compound target engagement and complex remodeling. This work establishes a foundational resource for decoding the regulatory proteome and provides a blueprint for integrating regulome data into next-generation models of cellular behavior.

cell biology↗

Ruxolitinib clears CRYAB p.Arg120Gly aggregates through the ubiquitin-proteasome system

RationaleProtein accumulation is a hallmark of many neurodegenerative and muscular diseases. Desmin-related (cardio-) myopathy (DRM), a well-studied model for cardiac muscle protein accumulation, is an autosomal dominant-inherited disease presenting with progressive muscle weakness, reduced quality of life, and shortened life span. To date, DRM patients are treated symptomatically and there is no causal treatment available. Independent of the genetic cause, most DRM patients display intracellular accumulation of desmin and its chaperone B-crystallin (CRYAB). We previously conducted an unbiased high-throughput screen to identify novel effectors that reduce cardiomyocyte aggregate levels and found that downregulation of Janus kinase 1 (JAK1) resulted in lower aggregate load in neonatal mouse cardiomyocytes. ObjectiveIn this study, we tested if the approved JAK inhibitor ruxolitinib ameliorates the disease phenotype in rodent and human CRYAB p.Arg120Gly DRM models. Methods and ResultsWe found that the mRNA levels of Jak1 and Stat3 were higher than any other JAK-signal transducer and activator of transcription (STAT) family members in neonatal rat ventricular myocytes (NRVMs) and human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). The approved JAK1/2 inhibitor ruxolitinib and the JAK1 inhibitors solcitinib, upadacitinib, and filgotinib prevented accumulation of and cleared pre-existing CRYAB p.Arg120Gly protein aggregates in NRVMs and hiPSC-CMs. Importantly, the knockdown of Jak1 and Stat3, but not Jak2 resulted in fewer aggregates. Moreover, ruxolitinib, Jak1 or Stat3 siRNA treatment enhanced the ubiquitin-proteasome system (UPS)-mediated degradation. Blocking UPS function blunted the effect of ruxolitinib or Jak1 siRNA on CRYAB p.Arg120Gly accumulation. RNAseq of NRVMs treated with Jak1 siRNA extracts revealed higher gene expression of important muscle E3 ubiquitinating enzymes. Knockdown of the E3 ligase Asb2 (Ankyrin Repeat And SOCS Box Protein 2) abolished the effect of ruxolitinib on CRYAB p.Arg120Gly aggregates. In DRM mice, phospho-STAT3 levels were markedly higher than in non-transgenic (NTG) mice with age. Ruxolitinib treatment or Jak1 knockout prevented cardiac dysfunction and reduced CRYAB p.Arg120Gly aggregate load in DRM mice. ConclusionIn this study, we uncovered the previously unknown effect of the approved drug ruxolitinib to enhance UPS-mediated degradation and prevent protein aggregates in cardiomyocytes.

pharmacology and toxicology↗

Effects of satellite-linked telemetry tags on humpback whales in the Gulf of Maine: photographic assessment of tag sites

Hundreds of large whales have been tracked using consolidated (Type-C) satellite tags, yet there have been few studies on their impacts on whale health. In 2011, we initiated the first study designed to evaluate the effects of these tags in a baleen whale. Between 2011 and 2018, we tagged 79 North Atlantic humpback whales in the Gulf of Maine. We initially deployed commonly-used tags with an articulation between the anchor and transmitter (n=35, 2011-2012). However, evidence of breakage prompted the development and use of more robust, integrated tags (n=45). Tagged individuals were photographed immediately prior to, during and up to 11 years after tagging. They were re-encountered on an average of 41.3 days (SD=44.3), yielding 2,971 photographed sightings through 2022. An objective scoring system was developed to characterise tag site tissue responses based on photographs and to identify risk factors for prolonged healing. The initial tissue response to tagging was minimal, followed by skin loss around the tag, sometimes a degree of swelling, occasional extrusion of blubber, changes in skin colour, local depression formation, tag loss and skin healing over the tag site, sometimes with a depression remaining. At last sighting, most non-integrated and integrated tag sites exhibited small shallow skin depressions (58.8% and 66.7%, respectively). Some exhibited deeper depressions with differing adjacent skin coloration (26.5% and 15.6%, respectively) or barely detectable marks (11.8% and 15.6%, respectively). Mild swellings occasionally persisted at the tag site, but this was uncommon for both tag designs (2.9% and 2.2%, respectively). More severe tissue responses were associated with non-integrated tags and placements lower on the body. This study highlights the importance of using robust tag designs to minimise negative effects from Type-C tags. Furthermore, because tag placement was shown to affect outcome, precision equipment, experienced taggers and vessel operators are critical for optimal deployments.

ecology↗

Before and after delisting: population dynamics of North Atlantic humpback whales over two decades in the Gulf of Maine

Whales are long-lived, slow-reproducing species that were decimated by commercial whaling. Although some populations seem close to recovery, others are challenging to assess. We studied humpback whales (Megaptera novaeangliae) in the Gulf of Maine, an area of the North Atlantic (NA) with well-documented threats. Long-term studies and mark-recapture data were used to estimate humpback whale apparent survival, abundance and population growth from 2000 through 2019. Estimates were derived from a hierarchical, Bayesian state-space model with sex, age, and random time effects on survival while accounting for individual capture probability. Abundance increased from 744 (95% CI: 726-762) in 2000 to 1,706 (95% CI: 1,639-1,771) in 2019, with 4.6% mean annual growth. However, adult males exhibited higher survival and outnumbered adult females by the end of the study. Over time, fewer calves were observed, calf survival varied and the juvenile class declined. These are rare insights into the dynamics underlying whale abundance trends and they revealed similarities to an endangered species that is declining due to environmental and human impacts. The results inform a listing change under the U.S. Endangered Species Act, a mortality event of unprecedented magnitude off the U.S and humpback whale recovery from historical whaling in the NA.

ecology↗

Caller identification and characterization of individual humpback whale acoustic behavior

Acoustic recording tags are biologging tools that provide fine scale data linking acoustic signaling with individual behavior; however, when an animal is in a group, it is challenging to tease apart calls of other conspecifics and identify which individuals produce each call. This, in turn, prohibits robust assessment of individual acoustic behavior including call rates and silent periods, call bout production within and between individuals, and caller location. To overcome this challenge, we simultaneously instrumented small groups of humpback whales on a western North Atlantic feeding ground with sound and movement recording tags. This simultaneous tagging approach enabled us to compare the relative amplitude of each call across individuals and infer caller identity though amplitude differences. Focusing on periods when the tagged animals were isolated from other conspecifics, we were able to assign caller ID for 97% of calls in this dataset. From these labeled calls, we found that humpback whale individual call rates are highly variable across individuals and groups (0-89 calls/h), with calls produced throughout the water column and in bouts with short inter-call intervals (ICI = 2.2 s). Most calls received a likely response from a conspecific within 100 s. These results are important for modelling signal detection range for passive acoustic monitoring and density estimation. Future studies can expand on these methods for caller identification and further investigate the nature of sequence production and counter-calling in humpback whale social calls. Finally, this approach can be helpful for understanding intra-group communication in social groups across other taxa. Summary statementTagging entire humpback whale social groups with sound and movement recording tags allows us to for the first time parse out call behavior within groups and understand individual acoustic behavior.

animal behavior and cognition↗

Universal baleen whale microsatellite panel for individual identification and power to detect parentage

Highly polymorphic single tandem repeat loci (STR, also known as microsatellite loci) remain a familiar, cost efficient class of markers for genetic analyses in ecology, behavior and conservation. We characterize a new universal set of ten STR loci (from 28 potential candidate loci) in seven baleen whale species, which are optimized for PCR amplification in two multiplex reactions along with a Y chromosome marker for sex determination. The optimized, universal set of STR loci provides an ideal starting point for new studies in baleen whales aimed at individual-based and population genetic studies, and facilitates data sharing among research groups. Data from the new STR loci were combined with genotypes from other published STR loci to assess the power to assign parentage (paternity) using exclusion in four species: fin whales, humpback whales, blue whales and bowhead whales. We argue that parentage studies should present a power analysis to demonstrate that the specific data are sufficiently informative to assign parentage with statistical rigor.

genetics↗

Direct estimation of genome mutation rates from pedigrees in free-ranging baleen whales

Current low germline mutation rate () estimates in baleen whales have greatly influenced research ranging from assessments of whaling impacts to evolutionary cancer biology. However, the reported rates were subject to methodological errors and uncertainty. We estimated directly from pedigrees in natural populations of four baleen whale species and the results were similar to primates. The implications of revised values include pre-exploitation population sizes at 14% of previous genetic diversity-based estimates and the conclusion that in itself is insufficient to explain low cancer rates in gigantic mammals (i.e., Petos Paradox). We demonstrate the feasibility of estimating from whole genome pedigree data in natural populations, which has wide-ranging implications for the many ecological and evolutionary inferences that rely on .

genetics↗

PHFinder: Assisted detection of point heteroplasmy in Sanger sequencingchromatograms

O_LIHeteroplasmy is the presence of two or more organellar genomes (mitochondrial or plastid DNA) in an organism, tissue, cell or organelle. Heteroplasmy can be detected by visual inspection of Sanger sequencing chromatograms, where it appears as multiple peaks of fluorescence at a single nucleotide position. Visual inspection of chromatograms is both consuming and highly subjective, as heteroplasmy is difficult to differentiate from background noise. Few software solutions are available to automate the detection of point heteroplasmies, and those that are available are typically proprietary, lack customization or are unsuitable for automated heteroplasmy assessment in large datasets. C_LIO_LIHere, we present PHFinder, a Python-based, open source tool to assist in the detection of point heteroplasmies in large numbers of Sanger chromatograms. PHFinder automatically identifies point heteroplasmies directly from the chromatogram trace data. The program was tested with Sanger sequencing data from 100 humpback whale (Megaptera novaeangliae) tissue samples with known heteroplasmies. C_LIO_LIPHFinder detected most (90%) of the known heteroplasmies thereby greatly reducing the amount of visual inspection required. PHFinder is flexible, enabling explicit specification of key parameters to infer double peaks (i.e., heteroplasmies). C_LI

bioinformatics↗

DNA methylation age studies of humpback whales

Several previous studies have described epigenetic clocks for whale and dolphin species. Here we present a novel and highly robust epigenetic clock for the humpback whale based on methylation levels measured using the Mammalian Methylation Array platform. Skin samples were obtained from 76 individuals that had been studied since their birth year and known to range in age from <1 to 39.5 years. The humpback whale clock provided a highly accurate estimate of chronological age (R=0.96, median error 2.2 years) according to a leave-one-out cross validation analysis. We applied this clock to an independent set of samples from humpback whales of unknown exact age but with sighting histories that were as long as or longer than the upper 20% of the available known-age range. Although there was a strong correlation with minimum age (R=0.89), the clock underestimated age in these older animals by a median error of at least 7.8 years. Finally, we applied the humpback clock to publicly available methylation data from beluga whales. In this data set from a different species, the humpback clock provided an age correlation of R=0.78. While a DNAm age estimator has previously been described for humpback whales, this is the first such clock shown to apply to another cetacean species as well. Our humpback whale clock built from well-studied population lends itself for understanding humpback populations that otherwise lack age data.

developmental biology↗

DNA Methylation Networks Underlying Mammalian Traits

Epigenetics has hitherto been studied and understood largely at the level of individual organisms. Here, we report a multi-faceted investigation of DNA methylation across 11,117 samples from 176 different species. We performed an unbiased clustering of individual cytosines into 55 modules and identified 31 modules related to primary traits including age, species lifespan, sex, adult species weight, tissue type and phylogenetic order. Analysis of the correlation between DNA methylation and species allowed us to construct phyloepigenetic trees for different tissues that parallel the phylogenetic tree. In addition, while some stable cytosines reflect phylogenetic signatures, others relate to age and lifespan, and in many cases responding to anti-aging interventions in mice such as caloric restriction and ablation of growth hormone receptors. Insights uncovered by this investigation have important implications for our understanding of the role of epigenetics in mammalian evolution, aging and lifespan.

evolutionary biology↗

Strong and lasting impacts of past global warming on baleen whale and prey abundance

AbstractThe demography of baleen whales and their prey during the past 30 thousand years was assessed to understand the effects of past rapid global warming on marine ecosystems. Mitochondrial and genome-wide DNA sequence variation in eight baleen whale and seven prey species revealed strong, ocean-wide demographic changes that were correlated with changes in global temperatures and regional oceanographic conditions. In the Southern Ocean baleen whale and prey abundance increased exponentially and in apparent synchrony, whereas changes in abundance varied among species in the more heterogeneous North Atlantic Ocean. The estimated changes in whale abundance correlated with increases in the abundance of prey likely driven by reductions in sea-ice cover and an overall increase in primary production. However, the specific regional oceanographic environment, trophic interactions and species ecology also appeared to play an important role. Somewhat surprisingly the abundance of baleen whales and prey continued to increase for several thousand years after global temperatures stabilized. These findings warn of the potential for dramatic, long-term effects of current climate changes on the marine ecosystem.\n\nOne Sentence SummaryThe effects of past global warming on marine ecosystems were drastic, system-wide and long-lasting.

evolutionary biology↗

Fin whale (Balaenoptera physalus) mitogenomics: A cautionary tale of defining sub-species from mitochondrial sequence monophyly

HighlightsO_LIMitochondrial monophyly is commonly employed to define evolutionary significant units.\nC_LIO_LIMonophyly may be caused by insufficient sampling or a recent common ancestor.\nC_LIO_LIMitogenomic studies are generally based on few samples and prone to sampling issues.\nC_LIO_LIExpanded mitogenome sampling negates previous monophyly in fin whales.\nC_LI\n\nAbstractThe advent of massive parallel sequencing technologies has resulted in an increase of studies based upon complete mitochondrial genome DNA sequences that revisit the taxonomic status within and among species. Spatially distinct monophyly in mitogenomic genealogies, i.e., the sharing of a recent common ancestor among con-specific samples collected in the same region has been viewed as evidence for subspecies. Several recent studies in cetaceans have employed this criterion to suggest subsequent intraspecific taxonomic revisions. We reason that employing intra-specific, spatially distinct monophyly at non-recombining, clonally inherited genomes is an unsatisfactory criterion for defining subspecies based upon theoretical (genetic drift) and practical (sampling effort) arguments. This point is illustrated by a re-analysis of a global mitogenomic assessment of fin whales, Balaenoptera physalus spp., published by Archer et al. (2013) which proposed to further subdivide the Northern Hemisphere fin whale subspecies, B. p. physalus. The proposed revision was based upon the detection of spatially distinct monophyly among North Atlantic and North Pacific fin whales in a genealogy based upon complete mitochondrial genome DNA sequences. The extended analysis conducted in this study (1,676 mitochondrial control region, 162 complete mitochondrial genome DNA sequences and 20 microsatellite loci genotyped in 358 samples) revealed that the apparent monophyly among North Atlantic fin whales reported by Archer et al. (2013) to be due to low sample sizes. In conclusion, defining sub-species from monophyly (i.e., the absence of para-or polyphyly) can lead to erroneous conclusions due to relatively \"trivial\" aspects, such as sampling. Basic population genetic processes (i.e., genetic drift and migration) also affect the time to most recent common ancestor and hence the probability that individuals in a sample are monophyletic.\n\n\n\nO_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=195 SRC=\"FIGDIR/small/488130v1_ufig1.gif\" ALT=\"Figure 1\">\nView larger version (28K):\norg.highwire.dtl.DTLVardef@1e38e54org.highwire.dtl.DTLVardef@9533eborg.highwire.dtl.DTLVardef@183b5b5org.highwire.dtl.DTLVardef@eae09_HPS_FORMAT_FIGEXP M_FIG C_FIG

evolutionary biology↗

Pisces: An Accurate and Versatile Variant Caller for Somatic and Germline Next-Generation Sequencing Data

MotivationNext-Generation Sequencing (NGS) technology is transitioning quickly from research labs to clinical settings. The diagnosis and treatment selection for many acquired and autosomal conditions necessitate a method for accurately detecting somatic and germline variants, suitable for the clinic.\n\nResultsWe have developed Pisces, a rapid, versatile and accurate small variant calling suite designed for somatic and germline amplicon sequencing applications. Pisces accuracy is achieved by four distinct modules, the Pisces Read Stitcher, Pisces Variant Caller, the Pisces Variant Quality Recalibrator, and the Pisces Variant Phaser. Each module incorporates a number of novel algorithmic strategies aimed at reducing noise or increasing the likelihood of detecting a true variant.\n\nAvailabilityPisces is distributed under an open source license and can be downloaded from https://github.com/Illumina/Pisces. Pisces is available on the BaseSpace SequenceHub as part of the TruSeq Amplicon workflow and the Illumina Ampliseq Workflow. Pisces is distributed on Illumina sequencing platforms such as the MiSeq, and is included in the Praxis Extended RAS Panel test which was recently approved by the FDA for the detection of multiple RAS gene mutations.\n\nContactpisces@illumina.com\n\nSupplementary informationSupplementary data are available online.

bioinformatics↗