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Hannah, R.

Publications and source records attributed to Hannah, R..

15 recordsLinked to original sources

Self-supervised learning for a gene program-centric view of cell states

Single-cell omics has extended the biological interrogation of cell state from examining the expression of individual genes to unbiased profiling of tens of thousands of genes at once. However, extracting biological insights from such high-dimensional data remains challenging. To enable downstream analyses, many computational approaches compress cell state into a single latent representation. This can obscure the structure of underlying gene programs (GP), defined as coordinated sets of biologically related genes, such as signalling pathway response modules or transcription factor targets. Here, we present Tripso, a self-supervised transformer deep learning model which learns multiple GP-specific embeddings from predefined GPs, while also enabling the discovery of novel, data-driven GPs. Tripso facilitates principled comparisons across development, disease, and experimental systems. Firstly, in a dataset of human hematopoietic cells spanning prenatal development through adulthood and aging and including newly generated data, Tripso resolved age-specific GP patterns, including elevated JAK-STAT activity in pediatric hematopoietic cells and postnatal shifts in IKZF1 GP activity during B cell differentiation. Secondly, leveraging Tripso GP embeddings and comparing in vivo to in vitro data, we hypothesized and experimentally validated that inhibition of the SEC61 translocon improved maintenance of hematopoietic stem cells in culture. Finally, Tripsos capacity for data-driven GP discovery revealed a previously uncharacterized tissue-resident memory T cell GP with increased activity in atopic dermatitis. Its spatial co-localization with sebaceous gland-associated immune niches was demonstrated in spatial transcriptomic and proteomic data. Thus, by moving beyond single embeddings of cellular states, Tripso enables interpretable and actionable discoveries, demonstrating how GP-centric modelling can generate hypotheses with substantial biomedical relevance. By anchoring cellular representations in meaningful GPs, Tripso establishes a principled and biologically grounded framework towards the development of interpretable virtual cell models.

bioinformatics↗

Predicting how perturbations reshape cellular trajectories with PerturbGen

A major challenge in biology is predicting how cells transition between states over time and how perturbations disrupt these transitions. Understanding such dynamics is critical for identifying interventions that reverse pathological programs or reprogram cells toward desired states. Although recent computational approaches can predict single-cell perturbation responses in silico, they cannot predict responses across dynamic cell trajectories, for example how early perturbations reconfigure later cell states. To address this gap, we introduce PerturbGen, a generative foundation model trained on over 100 million single-cell transcriptomes that predicts perturbation responses along cellular trajectories. PerturbGen predicts how genetic perturbation at source state shapes downstream states, alters gene programs and trajectories across time, for example in differentiation or disease progression. We apply PerturbGen to three newly generated multi-condition human single-cell datasets spanning immune responses, hematopoiesis and skin development. In an in vivo immune challenge, PerturbGen predicts that knocking out an IL1B signal in myeloid cells attenuates later cytokine-interferon programs, with downstream changes consistent with a reversal of IL-1{beta} stimulation signature. In hematopoiesis, anchoring perturbation-induced programs to human genetics enables simulation of monogenic blood disorders, recapitulates established disease-associated biology whilst systematically revealing lineage-specific programs, including in lineages where this was not previously possible. In skin organoids, PerturbGen predicted that Wnt activation enhances stromal differentiation recapitulating the trajectory observed in human prenatal skin, findings that were functionally validated by experimentally activating Wnt signaling. Together, PerturbGen extends modeling of gene perturbations from static to dynamic cellular systems. We envision PerturbGen enabling the creation of in silico, trajectory-aware perturbation atlases and virtual cells across diverse biological scenarios, supporting optimization of disease models and prioritization of candidate molecular interventions for therapeutic discovery.

bioinformatics↗

TMS-Evoked Corticospinal Beta Oscillations in Humans Recorded from Muscles

BackgroundTranscranial magnetic stimulation (TMS) can entrain oscillatory brain activity, offering a promising approach to study motor-related neural oscillations such as beta rhythms. However, how TMS-induced corticospinal oscillations are generated, propagated, and related to endogenous activity remains unclear, partly due to limitations in brain recording techniques. Recording muscle activity provides an alternative and physiologically grounded window into corticospinal dynamics. MethodsWe investigated whether subthreshold TMS over the motor cortex induces corticospinal oscillatory activity detectable in muscles, and whether these responses share neural generators with endogenous beta rhythms. Single-pulse subthreshold TMS was applied over the motor cortex in healthy participants while electromyography (EMG) was recorded from tonically active muscles. Stimulation intensity, coil orientation, and stimulation site were systematically varied. Concurrent electroencephalography (EEG) was used to assess cortical responses and corticomuscular transmission. In additional experiments, advanced EMG techniques were employed to track motor neuron pools and characterize how TMS-evoked oscillations are transmitted at the motor unit level. ResultsTMS elicited a robust and short-latency increase in beta-band activity in the EMG. The analysis of the elicited muscle responses and the comparison of results across different TMS configurations indicate that the beta responses resulted from activation of inhibitory interneurons in the targeted primary motor cortex. Importantly, the characteristics of cortico-muscular coherence and beta projection to the muscles indicate that the elicited beta responses with TMS have same cortical sources as endogenously generated beta activity. ConclusionsThese findings demonstrate that muscle recordings provide a sensitive and physiologically meaningful readout of TMS-induced corticospinal beta oscillations.

neuroscience↗

Global motor system suppression as the primary mechanism of human action stopping: challenging the pause-then-cancel model

The ability to stop a planned or ongoing action is fundamental to inhibitory control. A recent theory proposes that stopping involves two distinct phases: an initial global suppression of motor activity ("pause") followed by a selective cancellation of the targeted action. However, the necessity of a second "cancel" stage remains debated. We tested whether global suppression alone is sufficient to stop movement by analysing electromyography from task-relevant agonist and antagonist muscles, alongside transcranial magnetic stimulation measures of global motor suppression from task-irrelevant muscles, during a stop-signal task in adult human participants of both sexes. In Experiment 1, reanalysis of ballistic finger movements revealed that agonist muscle offset consistently preceded behavioural stopping, aligning with the time course of global suppression. In Experiment 2, we extended these findings to whole-arm reaching movements, demonstrating that global motor suppression persisted beyond the termination of muscle activity when stopping prevented movement initiation, but disengaged in time for antagonist activation used to interrupt movements once they had begun. These findings challenge the pause-then-cancel model, instead supporting a single-stage global suppression framework. They also suggest that the global suppression mechanism is not a rigid, top-down stopping mechanism but rather part of a broader motor control system that flexibly adjusts movement commands based on task demands. Significance StatementA recent theory suggests that stopping an action involves a two-stage process: first pausing movement execution, then selectively cancelling specific motor commands. Our study challenges this view, demonstrating that global motor suppression alone is sufficient to terminate motor commands. Using electromyography and transcranial magnetic stimulation during two distinct motor tasks, we show that suppression fully curtails agonist motor commands before behavioural stopping, without requiring a distinct cancellation step. These findings reshape our understanding of action stopping and provide a foundation for exploring the pathophysiology of movement and neuropsychiatric disorders associated with impaired behavioural suppression.

neuroscience↗

Distinct impacts of sodium channel blockers on the strength-duration properties of human motor cortex neurones

Voltage-gated sodium channels (VGSCs) are essential for regulating axonal excitability in the human brain. While sodium channel blockers are known to modulate neuronal excitability, their in vivo effects on the human cortex remain poorly understood. Here, we employed novel transcranial magnetic stimulation (TMS) measures to investigate the effects of sodium channel blockers, carbamazepine and lacosamide, on the strength-duration behaviour of human cortical neurones, serving as an index of sodium channel function. Our data showed that single doses of both medications elevated resting motor thresholds compared to placebo, indicating reduced excitability; however, their impacts varied according to TMS pulse width. Carbamazepine raised thresholds proportionally across all pulse widths, whereas lacosamide disproportionately influenced thresholds for long-duration pulses. Crucially, lacosamide reduced the strength-duration time constant and increased rheobase, while carbamazepine had minimal effects on both. These results reveal subtle and unexpected differences in cortical neurone behaviour following VGSC-blocking medication administration. Lacosamides response aligns with the proposed mechanism of sodium conductance blockade, while carbamazepines effects suggest distinct VGSC interactions or potential off-target effects. Our findings advance the understanding of VGSC-blocking medication interactions in the human cortex and underscore the importance of employing specific TMS measures to gain deeper insights into medication mechanisms of action in vivo. Such measures could serve as valuable adjuncts in medication development and patient monitoring, enhancing understanding of medication action in clinical settings.

neuroscience↗

Delayed Trp53 activation protects Dnmt3a-mutant hematopoietic stem cells from inflammatory attrition.

Hematopoietic stem cells (HSCs) accumulate somatic mutations over time, some conferring a fitness advantage that can lead to clonal hematopoiesis (CH). Mutations in DNMT3A, particularly at hotspot R882, are the most prevalent in CH and carry an increased risk of acute myeloid leukemia (AML). Although DNMT3A R882 mutations are linked to global DNA hypomethylation, the mechanisms underlying their selective advantage remain unclear. Here, we show that Dnmt3a-R882H mutant HSCs exhibit resilience under inflammatory and genotoxic stress. During IL-1{beta}-induced emergency granulopoiesis, Dnmt3a R882H/+ HSCs uncouple increased proliferation from stem cell exhaustion. In contrast, wild-type HSCs rapidly progress to terminal differentiation. We link this phenotype to a delayed activation of the p53-p21-DREAM axis, that allows mutant HSCs to avoid attrition, despite increased replication stress. Similarly, mutant HSCs exhibit delayed Trp53 activation following irradiation, but eventually recover a physiological Trp53 response. Analysis of patient data reveals shared phenotypic features between DNMT3A and monoallelic TP53 mutations in CH and myeloid neoplasms, highlighting potential functional similarities. Collectively, these findings suggest that the expansion of DNMT3A-mutant clones is affected by impaired TP53 signaling, which confers resilience against stressors. Therapeutic strategies targeting inflammatory pathways or the p53-p21-DREAM axis may reduce DNMT3A-CH expansion and/or progression and its associated risks.

molecular biology↗

Rate of force development is correlated with corticospinal excitability during explosive voluntary contractions.

ObjectiveTo investigate the relationship between rate of torque development (RTD) and corticospinal excitability (denoted by motor-evoked potential; MEP) during explosive voluntary contractions. Also, to assess differences in MEP and silent period duration between different phases of explosive contraction and at maximum voluntary contraction (MVC) plateau. MethodsIn 14 adults, quadriceps muscle MEP and silent period duration were measured at [~]45 (early), [~]115 (middle), and [~]190 ms (late) from EMG onset during knee-extensor isometric explosive contractions, and at MVC plateau with superimposed transcranial magnetic stimulation (TMS). RTD was measured immediately prior to early-phase MEP during the same contractions, and these two variables were correlated across separate contractions, within participants, via repeated measures correlation (RmCorr). RTD was also measured in explosive contractions without TMS over early-, middle-, and late-phases and correlated to MEP averaged across the corresponding and preceding phases, via Pearsons r correlations, assessing relationships across participants. MEP and silent period duration were compared (ANOVA) between the different phases and at MVC plateau. ResultsMEP and RTD were correlated across separate contractions within participants (RmCorr r=0.43), and in the middle phase across participants (Pearsons r=0.56). MEP and RTD were not correlated in other phases (r[≥]0.09). Silent period duration increased throughout the different phases of contraction and up to MVC plateau (ANOVA, p=0.001) but MEP remained constant (ANOVA, p=0.42). ConclusionThe correlations between MEP and RTD suggests corticospinal excitability is an important determinant of RTD. During rapid torque development and up to MVC plateau, corticospinal inhibition increases but excitability remains constant.

physiology↗

Test-retest reliability of TMS motor evoked responses and silent periods during explosive voluntary isometric contractions.

PurposeThis study assessed the intraclass correlation coefficient (ICC) and coefficient of variation (CV) test-retest reliability of TMS responses MEPs and silent periods at early, middle, and late phases of the rising time-torque curve during explosive voluntary contractions. We also investigated how the number of consecutively averaged measurements over 3 to 15 separate contractions influenced reliability. MethodsOn two separate occasions 3-7 days apart, 14 adults completed 48 isometric explosives (1-s) contractions of the knee extensors, superimposed with TMS. The TMS elicited an MEP and silent period in the quadriceps muscles at 45 (early), 115 (middle), or 190 ms (late) during each contraction. TMS was also superimposed at the plateau of 15 separate MVCs. Test-retest ICC and CV were calculated for consecutively averaged MEPs and silent periods. ResultsNo one condition was more reliable than another. For MEP amplitude, in all conditions except the explosive late phase, ICCs generally increased, and CV decreased, with an increase in the number of averaged contractions, and were >0.50 ICC and <15% CV within 7 contractions. For silent period, ICCs and CVs were unaffected by the number of consecutively averaged contractions and remained >0.50 ICC and <10% CV. ConclusionTest-retest reliability of TMS responses is comparable between phases of explosive contraction and at the plateau of MVC. To enhance MEP reliability, with the increases he number of averaged contractions of except the explosive late phase, we recommend studies average data across > the 3-5 contractions typically reported in the literature for studies using TMS during MVCs.

physiology↗

Maintenance of haematopoietic stem cells by JAK inhibition and increased tyrosine-unphosphorylated STAT5

Normal and malignant hematopoietic stem cells (HSCs) are controlled by extracellular cues including cytokine signalling through the JAK/STAT pathway. Here, we show that STAT5-deficient HSCs exhibit an unusual phenotype: while reduced multi-lineage repopulation and reduced self-renewal are commonly associated with overproliferation and exhaustion, they are instead associated with reduced cell-cycle progression and increased differentiation in STAT5-deficient HSCs. Mechanistic studies show that unphosphorylated-STAT5 (uSTAT5) contributes to this phenotype by constraining HSC differentiation, promoting HSC maintenance and upregulating transcriptional programs associated with stemness. The JAK1/2 inhibitor ruxolitinib increases levels of uSTAT5, constrains differentiation and proliferation of murine HSCs, promotes their maintenance and upregulates transcriptional programs associated with stemness. Ruxolitinib also enhances clonogenicity of normal human HSPCs, CALR-mutant murine HSCs and HSPCs from patients with myelofibrosis. Our results therefore reveal a previously unrecognized role for uSTAT5 in controlling HSC function, highlight JAK inhibition as a strategy for enhancing HSC function and provide insights into the failure of JAK inhibitors to eradicate myeloproliferative neoplasms.

cell biology↗

Multi-organ functions of yolk sac during human early development

The yolk sac (YS) represents an evolutionarily-conserved extraembryonic structure that ensures timely delivery of nutritional support and oxygen to the developing embryo. However, the YS remains ill-defined in humans. We therefore assemble a complete single cell 3D map of human YS from 3-8 post conception weeks by integrating multiomic protein and gene expression data. We reveal the YS as a site of primitive and definitive haematopoiesis including a YS-specific accelerated route to macrophage production, a source of nutritional/metabolic support and a regulator of oxygen-carrying capacity. We reconstruct the emergence of primitive haematopoietic stem and progenitor cells from YS hemogenic endothelium and their decline upon stromal support modulation as intraembryonic organs specialise to assume these functions. The YS therefore functions as three organs in one revealing a multifaceted relay of vital organismal functions as pregnancy proceeds. One Sentence SummaryHuman yolk sac is a key staging post in a relay of vital organismal functions during human pregnancy.

developmental biology↗

Temporally-precise disruption of prefrontal cortex informed by the timing of beta bursts impairs human action-stopping

Human action-stopping is thought to rely on a prefronto-basal ganglia-thalamocortical network, with right inferior frontal cortex (rIFC) posited to play a critical role in the early stage of implementation. Here we sought causal evidence for this idea in experiments involving healthy human participants. We first show that action-stopping is preceded by bursts of electroencephalographic activity in the beta band over prefrontal electrodes, putatively rIFC, and that the timing of these bursts correlates with the latency of stopping at a single-trial level: earlier bursts are associated with faster stopping. From this we reasoned that the integrity of rIFC at the time of beta bursts might be critical to successful stopping. We then used fMRI-guided transcranial magnetic stimulation (TMS) to disrupt rIFC at the approximate time of beta bursting. Stimulation prolonged stopping latencies and, moreover, the prolongation was most pronounced in individuals for whom the pulse appeared closer to the presumed time of beta bursting. These results help validate a model of the neural architecture and temporal dynamics of action-stopping. They also highlight the usefulness of prefrontal beta bursts to index an apparently important sub-process of stopping, the timing of which might help explain within- and between-individual variation in impulse control.

neuroscience↗

Temporal cascade of frontal, motor and muscle processes underlying human action-stopping

Action-stopping is a canonical executive function thought to involve top-down control over the motor system. Here we aimed to validate this stopping system using high temporal resolution methods in humans. We show that, following the requirement to stop, there was an increase of right frontal beta ([~]13 to 30 Hz) at [~]120 ms, likely a proxy of right inferior frontal gyrus; then, at 140 ms, there was a broad skeletomotor suppression, likely reflecting the impact of the subthalamic nucleus on basal ganglia output; then, at [~]160 ms, suppression was detected in the muscle, and, finally, the behavioral time of stopping was [~]220 ms. This temporal cascade confirms a detailed model of action-stopping, and partitions it into subprocesses that are isolable to different nodes and are more precise than the behavioral speed of stopping. Variation in these subprocesses, including at the single-trial level, could better explain individual differences in impulse control.

neuroscience↗

Excitability changes in the motor cortex in preparation for self-paced and cue-guided movements: a transcranial magnetic stimulation study

In a warned reaction time (RT) task, corticospinal excitability (CSE) decreases in task-related muscles at the time of the imperative signal (preparatory inhibition). Because RT tasks emphasise speed of response, it is impossible to distinguish whether preparatory inhibition reflects a mechanism preventing premature reactions, or whether it is an inherent part of movement preparation. We used transcranial magnetic stimulation (TMS) to study CSE changes preceding RT movements and movements that were either self-paced (SP) or performed at predictable times to coincide with an external event (PT). Results show that CSE changes over a similar temporal profile in all cases, suggesting that preparatory inhibition is a necessary state in planned movements allowing the transition between rest and movement. Additionally, TMS given shortly before the times to move speeded the onset of movements in both RT and SP contexts, suggesting that their initiation depends on a form of trigger that can be conditioned by external signals. On the contrary, PT movements do not show this effect, suggesting the use of a mechanistically different triggering strategy. This relative immunity of PT tasks to be biased by external events may reflect a mechanism that ensures priority of internal predictive signals to trigger movement onset.

neuroscience↗

Characterizing Genetic Circuit Components in E. coli towards a Campylobacter jejuni Biosensor.

Campylobacter jejuni is responsible for most cases of bacterial gastroenteritis (food poisoning) in the United Kingdom. The most common routes of transmission are by contact with raw poultry. Current detection systems for the pathogen are time-consuming, expensive or inaccessible for everyday users. In this article we propose a cheaper and faster system for detection of C. jejuni using a synthetic biology approach. We aimed to detect C. jejuni by the presence of xylulose, an uncommon bacterial capsular saccharide. We characterized two sugar-based regulatory systems that displayed potential to act as tools for detection of xylulose. Using a two-plasmid reporter system in Escherichia coli, we investigated the regulatory protein component (MtlR) of the mannitol operon from Pseudomonas fluorescens. Our findings suggest that the promoter of mtlE is activated by MtlR in the presence of a variety of sugar inducer molecules, and may exhibit cross-activity with a native regulator of E. coli. Additionally, we engineered the L-arabinose transcriptional activator (AraC) of E. coli for altered ligand specificity. We performed site-specific saturation mutagenesis to generate AraC variants with altered effector specificity, with an aim to generate a mutant activated by xylulose. We characterized several mutant AraC variants which have lost the ability to respond specifically to the native L-arabinose effector. We promote this technique as a powerful tool for future iGEM teams to create regulatory circuits activated by novel small molecule ligands.

synthetic biology↗

Effects of coil orientation on Motor Evoked Potentials from Orbicularis Oris and First Dorsal Interosseus

ObjectiveThis study aimed to characterise effects of coil orientation on the size of Motor Evoked Potentials (MEPs) from both sides of Orbicularis Oris (OO) and compare these effects with those reported for First Dorsal Interosseous (FDI), following stimulation to left lip and left hand Primary Motor Cortex.\n\nMethodsUsing a 70 mm figure-of-eight coil, we collected MEPs from eight different orientations while recording from contralateral and ipsilateral OO and FDI using a monophasic pulse.\n\nResultsMEPs from OO were evoked consistently for six out of eight orientations for contralateral and ipsilateral sites. When latency and silent periods were taken into account, contralateral orientations 0{degrees}, 45{degrees}, 90{degrees}, and 315{degrees} were found to best elicit OO MEPs with a likely cortical origin. As expected, the largest FDI MEPs were recorded with an orientation of 45{degrees}, invoking a posterior-anterior (PA) current flow, from the contralateral location.\n\nConclusionOrientations traditionally used for FDI were also found suitable for eliciting OO MEPs. Individuals vary more in their optimal coil orientation for eliciting MEPs from OO than for FDI. It is recommended that researchers iteratively probe several orientations when eliciting MEPs from OO. Care must be taken however because several orientations likely induced direct activation of facial muscles.

neuroscience↗