Biology subjects

Davies, J.

Publications and source records attributed to Davies, J..

68 records · Page 4Linked to original sources

bioRxiv2020

LY-CoV555, a rapidly isolated potent neutralizing antibody, provides protection in a non-human primate model of SARS-CoV-2 infection

SARS-CoV-2 poses a public health threat for which therapeutic agents are urgently needed. Herein, we report that high-throughput microfluidic screening of antigen-specific B-cells led to the identification of LY-CoV555, a potent anti-spike neutralizing antibody from a convalescent COVID-19 patient. Biochemical, structural, and functional characterization revealed high-affinity binding to the receptor-binding domain, ACE2 binding inhibition, and potent neutralizing activity. In a rhesus macaque challenge model, prophylaxis doses as low as 2.5 mg/kg reduced viral replication in the upper and lower respiratory tract. These data demonstrate that high-throughput screening can lead to the identification of a potent antiviral antibody that protects against SARS-CoV-2 infection. One Sentence SummaryLY-CoV555, an anti-spike antibody derived from a convalescent COVID-19 patient, potently neutralizes SARS-CoV-2 and protects the upper and lower airways of non-human primates against SARS-CoV-2 infection.

immunology↗

bioRxiv2020

Enhanced BCR signalling inflicts early plasmablast and germinal centre B cell death

It is still not clear how B-cell receptor (BCR) signalling intensity affects plasma cell and germinal centre (GC) B cell differentiation. We generated C{gamma}1Cre/+Ptpn6fl/fl mice where SHP-1, a negative regulator of BCR signalling, is deleted rapidly after B cell activation. Although immunisation with T-dependent antigens increased BCR signalling, it led to plasma cells reduction and increased apoptosis. Dependent on the antigen, the early GC B cell response was equally reduced and apoptosis increased. At the same time, a higher proportion of GC B cells expressed cMYC, indicating increased GC B cell - Tfh cell interactions. While GC B cell numbers returned to normal at later stages, affinity maturation was suppressed in the long term. This confirms that BCR signalling not only directs affinity dependent B cell selection but also, without adequate Tfh cell help, can inflict cell death, which may be important for the maintenance of B cell tolerance.

immunology↗

bioRxiv2020

Opposing macroevolutionary and trait-mediated patterns of threat and naturalization in flowering plants

Due to expanding global trade and movement, new plant species are establishing in exotic ranges at increasing rates while the number of native species facing extinction from multiple threats grows. Yet, how species losses and gains globally may together be linked to traits and macroevolutionary processes is poorly understood. Here we show that, adjusting for diversification rate and age, the proportion of threatened species across flowering plant families is negatively related to the proportion of naturalized species. Moreover, naturalization is positively associated with climate variability, short generation time, autonomous seed production, and interspecific hybridization, but negatively with age and diversification; whereas threat is negatively associated with climate variability and hybridization, and positively with biotic pollination, age and diversification. Such a pronounced signature of naturalization and threat across plant families suggests that both trait syndromes have coexisted over deep evolutionary time and that neither strategy is necessarily superior to the other.

ecology↗

bioRxiv2020

The fiber knob protein of human adenovirus type 49 mediates highly efficient and promiscuous infection of cancer cell lines using a novel cell entry mechanism

The human adenovirus (HAdV) phylogenetic tree is diverse, divided across seven species and comprising over 100 individual types. Species D HAdV are rarely isolated with low rates of pre-existing immunity, making them appealing for therapeutic applications. Several species D vectors have been developed as vaccines against infectious diseases where they induce robust immunity in pre-clinical models and early phase clinical trials. However, many aspects of the basic virology of species D HAdV, including their basic receptor usage and means of cell entry, remain understudied. Here, we investigated HAdV-D49, which previously has been studied for vaccine and vascular gene transfer applications. We generated a pseudotyped HAdV-C5 presenting the HAdV-D49 fiber knob protein (HAdV-C5/D49K). This pseudotyped vector was efficient at infecting cells devoid of all known HAdV receptors, indicating HAdV-D49 uses an unidentified cellular receptor. Conversely, a pseudotyped vector presenting the fiber knob protein of the closely related HAdV-D30 (HAdV-C5/D30K), differing in four amino acids to HAdV-D49, failed to demonstrate the same tropism. These four amino acid changes resulted in a change in isoelectric point of the knob protein, with HAdV-D49K possessing a basic apical region compared to a more acidic region in HAdV-D30K. Structurally and biologically we demonstrate that HAdV-D49 knob protein is unable to engage CD46, whilst potential interaction with CAR is extremely limited by extension of the DG loop. HAdV-C5/49K efficiently transduced cancer cell lines of pancreatic, breast, lung, oesophageal and ovarian origin, indicating it may have potential for oncolytic virotherapy applications, especially for difficult to transduce tumour types. ImportanceAdenoviruses are powerful tools experimentally and clinically. To maximise efficacy, the development of serotypes with low pre-existing levels of immunity in the population is desirable. Consequently, attention has focussed on those derived from species D, which have proven robust vaccine platforms. This widespread usage is despite limited knowledge in their basic biology and cellular tropism. We investigated the tropism of HAdV-D49, demonstrating it uses a novel cell entry mechanism that bypasses all known HAdV receptors. We demonstrate, biologically, that a pseudotyped HAdV-C5/D49K vector efficiently transduces a wide range of cell lines, including those presenting no known adenovirus receptor. Structural investigation suggests that this broad tropism is the result of a highly basic electrostatic surface potential, since a homologous pseudotyped vector with a more acidic surface potential, HAdV-C5/D30K, does not display a similar pan-tropism. Therefore, HAdV-C5/D49K may form a powerful vector for therapeutic applications capable of infecting difficult to transduce cells.

microbiology↗

bioRxiv2020

The effect of delayed processing on ovarian tissue stored for fertility preservation

BackgroundOvarian tissue cryopreservation (OTC) is important for fertility preservation and conservation. Delay in OTC may be required for transport or workflow management, however little is understood about the effect of processing delay on the tissue. ObjectiveTo determine whether a delay of 24-48 hours to OTC affects primordial follicle (PF) health. MethodsOvaries (n=6 sheep) were processed immediately or after storage at 4{degrees}C (24h, 48h). Tissue was fixed fresh, after cryopreservation or 10-day xenotransplantation. Morphological assessment of follicle health and development was performed. FindingsA total of 1541 follicles were analysed. A 24h processing delay did not impact PF health in fresh or cryopreserved tissue. In fresh tissue a 48h delay had an adverse effect on follicle health (OR=2.47, 95% CI 1.29-4.71). Interestingly, a 48h delay resulted in cryopreserved tissue being less likely to be graded as unhealthy compared to control (OR=0.56, 95% CI 0.36-0.87). There was no difference in PF health or development across groups following xenotransplantation. ConclusionOvarian tissue can be stored for up to 48 hours prior to cryopreservation with no net impact on PF health.

physiology↗

bioRxiv2020

Recalibrating the Epigenetic Clock: Implications for Assessing Biological Age in the Human Cortex

Human DNA-methylation data have been used to develop biomarkers of ageing - referred to epigenetic clocks - that have been widely used to identify differences between chronological age and biological age in health and disease including neurodegeneration, dementia and other brain phenotypes. Existing DNA methylation clocks are highly accurate in blood but are less precise when used in older samples or on brain tissue. We aimed to develop a novel epigenetic clock that performs optimally in human cortex tissue and has the potential to identify phenotypes associated with biological ageing in the brain. We generated an extensive dataset of human cortex DNA methylation data spanning the life-course (n = 1,397, ages = 1 to 104 years). This dataset was split into training and testing samples (training: n = 1,047; testing: n = 350). DNA methylation age estimators were derived using a transformed version of chronological age on DNA methylation at specific sites using elastic net regression, a supervised machine learning method. The cortical clock was subsequently validated in a novel human cortex dataset (n = 1,221, ages = 41 to 104 years) and tested for specificity in a large whole blood dataset (n = 1,175, ages = 28 to 98 years). We identified a set of 347 DNA methylation sites that, in combination optimally predict age in the human cortex. The sum of DNA methylation levels at these sites weighted by their regression coefficients provide the cortical DNA methylation clock age estimate. The novel clock dramatically out-performed previously reported clocks in additional cortical datasets. Our findings suggest that previous associations between predicted DNA methylation age and neurodegenerative phenotypes might represent false positives resulting from clocks not robustly calibrated to the tissue being tested and for phenotypes that become manifest in older ages. The age distribution and tissue type of samples included in training datasets need to be considered when building and applying epigenetic clock algorithms to human epidemiological or disease cohorts.

genomics↗

bioRxiv2019

Single cell transcriptomic analysis identifies Langerhans cells immunocompetency is critical for IDO1- dependent ability to induce tolerogenic T cells.

Human epidermal Langerhans cells (LCs) can coordinate both immunogenic and tolerogenic immune responses, creating an attractive opportunity for immunomodulation strategies. To investigate transcriptional determinants of human primary LC tolerance we applied single cells RNA-sequencing combined with transcriptional network modelling and functional analysis. Unsupervised clustering of single cell transcriptomes revealed that steady-state LCs exist in immature and immunocompetent states, and become fully immunocompetent on migration. Interestingly, LC migration, which has been shown to result in upregulation of the transcription factor IRF4, led in parallel to increased expression of a tolerogenic gene module including IDO1, LGALS1, LAMTOR1 and IL10RA, which translated to efficient induction of regulatory T cells in co-culture assays by immunocompetent LCs. Using protein expression analysis and perturbation with inhibitors, we confirmed the role of IDO1 as a mediator of LC tolerogenic responses induced during LC migration. Computational analysis of regulons and Partial Information Decomposition analyses identified IRF4 as a key driver for LC tolerogenic programmes. The predicted IRF4-regulated genes were confirmed by analysis of CRISPR-Cas9 edited LCs. These findings suggest that efficient priming of tolerogenic responses by LCs requires upregulation of a migration-coupled maturation program which is superimposed with a tolerance-inducing genomic module.

systems biology↗

bioRxiv2019

Resolving cellular systems by ultra-sensitive and economical single-cell transcriptome filtering

Single-cell transcriptomics has sensitivity limits that restrict low abundance transcript identification, affects clustering and introduce artefact. Here, we describe Constellation DropSeq (C-DropSeq), a molecular transcriptome filter that delivers two orders of magnitude sensitivity gains by maximising read utility while reducing sequencing depth and costs. The simple and powerful method is broadly compatible with library preparation routines and was demonstrated by identifying and characterizing the activation of rare dendritic cell sub-populations.

molecular biology↗

bioRxiv2018

Data from an electronic health informatics pipeline to describe clearance dynamics of Hepatitis B surface antigen (HBsAg) and e-Antigen (HBeAg) in chronic HBV infection

HBsAg and HBeAg have gained traction as biomarkers of control and clearance during monitoring of chronic hepatitis B virus infection (CHB). An improved understanding of the correlates of clearance of these proteins could help inform improvements in patient-stratified care and advance insights into the underlying mechanisms of disease control, thus underpinning new cure strategies. We collected electronic clinical data via an electronic pipeline supported by the National Institute for Health Research Health Informatics Collaborative (NIHR-HIC), adopting an unbiased approach to generating a robust longitudinal dataset for adults testing HBsAg-positive from a large UK teaching hospital over a six year period (2011-2016 inclusive). From 553 individuals with CHB, longitudinal data were available for 319, representing >107,000 weeks of clinical follow-up. Among these 319 individuals, 13 (4%) cleared HBsAg completely. HBsAg clearance rate was similar in individuals on NA therapy (n=4, median clearance time 150 weeks) vs those not on NA therapy (n=9, median clearance time 157 weeks). Those who cleared HBsAg were significantly older, and less likely to be on NA therapy compared to non-clearers (p=0.003 and p=0.001, respectively). Chinese ethnicity was associated with HBeAg positivity (p=0.025). HBeAg clearance occurred both on NA therapy (n=24, median time 49 weeks) and off NA therapy (n=19, median time 52 weeks). Improved insights into the dynamics of these biomarkers can underpin better prognostication and patient-stratified care. Our systematised approach to data collection paves the way for scaling up efforts to harness clinical data to address research questions and underpin improvements in clinical care provision.\n\nIMPORTANCEAdvances in the diagnosis, monitoring and treatment of hepatitis B virus (HBV) infection are urgently required if we are to meet international targets for elimination by the year 2030. Here we demonstrate how routine clinical data can be harnessed through an unbiased electronic pipeline, showcasing the significant potential for amassing large clinical datasets that can help to inform advances in patient care, and provide clues that inform new cure strategies. Our cohort from a large UK hospital includes adults from diverse ethnic groups that have previously been under-represented in the literature. Tracking two protein biomarkers that are used to monitor chronic HBV infection, we provide new insights into the timelines of HBV clearance, both on and off treatment. These results contribute to improvements in individualised clinical care and may provide important clues into the immune events that underpin disease control.

microbiology↗

bioRxiv2018

Disease mortality in domesticated animals is predicted by host evolutionary relationships

Infectious diseases of domesticated animals impact human well-being via food insecurity, loss of livelihoods, and human infections. While much research has focused on parasites that infect single host species, most parasites of domesticated mammals infect multiple species. The impact of multi-host parasites varies across hosts; some rarely result in death, whereas others are nearly always fatal. Despite their high ecological and societal costs, we currently lack theory for predicting the lethality of multi-host parasites. Here, using a global dataset of over 4000 case-fatality rates for 65 infectious diseases (caused by micro and macro-parasites) and 12 domesticated host species, we show that the average evolutionary distance from an infected host to other mammal host species is a strong predictor of disease-induced mortality. We find that as parasites infect species outside of their documented phy-1 logenetic host range, they are more likely to result in lethal infections, with the odds of death doubling for each additional 10 million years of evolutionary distance. Our results for domesticated animal diseases reveal patterns in the evolution of highly lethal parasites that are difficult to observe in the wild, and further suggest that the severity of infectious diseases may be predicted from evolutionary relationships among hosts.

evolutionary biology↗

bioRxiv2018

The interaction of phylogeny and community structure: linking clades’ ecological structures and trait evolution

1AimCommunity phylogenetic studies use information about species evolutionary relationships to understand the processes of community ecological assembly. A central premise of the field is that species evolution maps onto ecological patterns, and phylogeny reveals something more than species traits alone. We argue, therefore, that there is a need to better understand and model the interaction of phylogeny with species traits and community composition.\n\nInnovationWe outline a new method that identifies clades with unusual ecological structures, based around partitioning the variation of species site occupancies ({beta}-diversity). Eco-phylogenetic theory would predict that these clades should also demonstrate distinct evolutionary trajectories. We suggest that modelling the evolution of independent trait data in these clades represents a strong test of whether there is an association between species ecological structure and evolutionary history.\n\nMain conclusionsUsing an empirical dataset of mammals from around the world, we identify two clades of rodents that tend not to co-occur (are phylogenetically overdispersed), and then find independent evidence of slower rates of body mass evolution in these clades. We suggest that our approach, which assumes nothing about the mode of species trait evolution but rather seeks to explain it using ecological information, presents a new way to examine eco-phylogenetic structure.

evolutionary biology↗

bioRxiv2017

Mammal extinctions and the increasing isolation of humans on the tree of life

A sixth great mass extinction is ongoing due to the direct and indirect effects of human pressures. However, not all lineages are impacted equally. As humans, we frequently believe that we hold a unique place on Earth. Here, we show that our current impacts on the natural world risk to heighten that expectation. Evolutionary proximity to Homo sapiens emerges as a powerful predictor of extinction risk among mammals. Our analysis shows that the species most closely related to H. sapiens are exposed to a large variety of threat types and that they may also have greater intrinsic sensitivity to threats. Pruning back the tree of life around us will lead to our species being among those with the fewest close relatives. We will erase our evolutionary history, forcing its uniqueness. If no action is taken, we will lose crucial biodiversity for the preservation of Earth ecosystems, and a key reference to what makes us human.

ecology↗

bioRxiv2017

A Liquid Chromatography-Mass Spectrometry Method For Screening Disulfide Tethering Fragments

We report the refinement of a high-throughput, liquid-chromatography/mass spectrometry (LC/MS)-based screening method for the identification of covalent small-molecule binders to proteins. Using a custom library of 1600 disulfide-capped fragments targeting surface cysteine residues, we optimize sample preparation, chromatography, and ionization conditions to maximize the reliability and flexibility of the approach. Data collection at a rate of 90 seconds per sample balances speed and reliability for sustained screening over multiple, diverse projects run over a 24-month period. The method is applicable to protein targets of various classes and a range of molecular masses. Data are processed in a custom pipeline that calculates a % bound value for each compound and detects false-positives by calculating significance of detected masses ( signal significance). An example pipeline has been made available through Biovias ScienceCloud Protocol Exchange. Data collection and analysis methods for the screening of covalent adducts of intact proteins are now fast enough to screen the largest covalent compound libraries in 1-2 days.

biophysics↗

bioRxiv2016

First Indications for Long-Term Benzodiazepine and Z-drugs use in the United Kingdom

Benzodiazepines and Z-drugs (BZDs), hypnotic drugs used for insomnia and anxiety, are prescribed millions of times a year in the UK. Although guidance from the relevant regulatory authorities (NICE and BNF) indicates them only for short-term use, the evidence suggests that many patients have been taking these drugs for much longer, often for decades. At present, there are no up-to-date, evidence-based estimates of the scale of long-term BZD use in the UK, which has prevented making a strong case for the need for withdrawal services. However, data obtained recently on BZD use from a number of GP surgeries (covering nearly 100,000 registered patients) in the North of England, allow such projections to be calculated. Scaling the results to a national level suggests that there are over a quarter of a million patients in the UK using BZDs for periods far longer than recommended. The projections also suggest that nearly half this number may be willing to accept help to stop their dependency on BZDs. These results indicate a serious problem, which should be addressed by more research into the harms associated with long-term BZD use, the provision of withdrawal services, and a national helpline to support patients with BZD dependency.

pharmacology and toxicology↗