Biology subjects

Evans, C.

Publications and source records attributed to Evans, C..

At least 19 recordsLinked to original sources

bioRxiv2026

Color-dependent foraging in C. elegans integrates chromoprotein photosensitization with bacterial metabolic cues

Animals rely on color to navigate complex environments, yet how eyeless organisms use chromatic information to guide food choice remains poorly understood. Here, we show that Caenorhabditis elegans exhibits robust color dependent foraging driven by microbial chromophores, preferentially consuming red while avoiding blue chromoprotein expressing bacteria across bacterial backgrounds and wild isolates. This discrimination persists in darkness and independently of photoreceptor, revealing a mechanism beyond canonical photoreception. Purified chromoproteins and bacterial metabolite fractions independently reproduce preference, demonstrating complementary chromatic and post ingestive metabolic cues. Mechanistically, blue chromoproteins generate singlet oxygen, producing oxidative stress and remodeling bacterial tryptophan and pterin metabolism, whereas red food promotes serotonin production and feeding-associated neuropeptide signaling. Disrupting serotonin biosynthesis or neuropeptide processing abolishes color preference. Together, our findings reveal a previously unrecognized, novel sensory strategy in which wavelength-selective pigment photochemistry transforms microbial color into metabolic information that is integrated through gut brain neuroendocrine signaling to guide foraging behavior in an eyeless animal.

molecular biology↗

bioRxiv2026

Spatially pooling photon information enables photon-efficient quantitative imaging

Quantitative fluorescence imaging techniques such as fluorescence lifetime imaging microscopy and hyperspectral imaging infer molecular contrast from photons distributed across spatial pixels and temporal or spectral channels. In the few-photon regime, however, conventional pixel-wise analysis discards the spatial relationships imposed across neighboring pixels by the microscope point-spread function (PSF). Here we show that this spatially distributed information can be recovered without prior knowledge of emitter positions, spatial support or component assignments. We introduce SPOOL (Spatially Pooled Optical Observation Likelihood), a training-free Poisson inverse framework that jointly recovers source-space amplitudes and quantitative contrast by combining the PSF with temporal-decay or spectral-response dictionaries. For an isolated source, the attainable precision gain is governed by a dimensionless optical quantity: the PSF width expressed in detector pixels. The predicted gain therefore scales with optical sampling rather than with the physical origin of the contrast. The model predicts that lifetime-precision gain scales approximately linearly with the number of pixels spanning the PSF full width at half maximum, a scaling reproduced by Monte Carlo simulations. At one detected photon per foreground pixel, the reconstruction reduces lifetime dispersion sixfold in fluorescent-bead experiments and decreases the lifetime root-mean-square error relative to a high-photon reference from 1.19 to 0.45 ns in dual-labeled cells. The same framework transfers unchanged to hyperspectral imaging, recovering spectral contrast from generic emission bands without prior fluorophore spectra.

biophysics↗

bioRxiv2026

SMART-NeuroDx: A Reagent-Free Multianalyte Biosensor Platform with Machine-Learning Readout for Point-of-Care Dementia Screening

Differentiating overlapping dementia pathologies, such as frontotemporal dementia and Alzheimers disease, calls for the simultaneous measurement of several blood biomarkers, yet electrochemical sensors remain predominantly single-target and dependent on labels and sample preparation. Here we report SMART-NeuroDx, a reagent-free electrochemical array that quantifies pTau217, GFAP, pTau181, and NfL directly from unprocessed plasma and serum in under 35 minutes. Four surface-confined redox-active molecularly imprinted polymer and aptamer-MIP recognition matrices are electropolymerized onto a four-working-electrode porous-gold printed-circuit array, using potential-assisted electrostatic gating keyed to each targets isoelectric point to prevent cross-channel template contamination during synthesis. Label-free Faradaic responses from the redox-active polymer backbone are acquired on a custom battery-powered STM32G4 handheld potentiostat and converted to concentration from five voltammetric features using cross-validated Random Forest and XGBoost regressors. The handheld unit reproduced the baseline fidelity of a commercial benchtop workstation and resolved sub-picogram pTau217 (limit of detection 0.087 pg mL-{superscript 1}) across marker-appropriate dynamic ranges, with inter-chip relative standard deviation at or below 5.34% and coefficients of determination of 0.94 to 0.99 against reference concentrations. Each channel retained selectivity in plasma and serum against competing neurological and inflammatory proteins, with non-specific signal deviation held below 10% by the hydrated PyPEG interfacial shell. The platform establishes reagent-free, simultaneous four-analyte neurodegeneration sensing on a single point-of-care device.

bioengineering↗

bioRxiv2026

Cofilin controls actin network identity by sorting actin binding proteins to distinct cytoskeletal structures

Proper cell physiology requires the co-assembly of multiple actin cytoskeletal networks that are tailored for specific functions. To maintain and promote the different functions of these networks, cells decorate them with distinct types of actin binding proteins (ABPs). While various models have been proposed to explain this selective sorting of ABPs, the role of actin disassembly factors is less well understood. Here, we used inducible CRISPR interference and quantitative live-cell imaging to test how disassembly factors control the ABP composition of different networks. We found that knockdown of cofilin (Cof1), a potent and highly conserved disassembly factor, disrupts the size, organization, and ABP composition of actin networks. Specifically, defects in Cof1-mediated disassembly disrupt intracellular transport due to the assembly of overgrown and disordered branched actin networks that are inappropriately decorated by tropomyosin (Tpm1). Contrary to prevailing models of ABP sorting, these networks are co-decorated by Tpm1 and fimbrin (Sac6), and their assembly is independent of formin activity. Instead, our findings support a model wherein failure to maintain the proper architecture of branched actin networks drives mis-localization of network-specific ABPs. Together, this work demonstrates that actin disassembly factors play a critical role in maintaining cytoskeletal structure and function to regulate ABP sorting across distinct networks.

Cell Biology↗

bioRxiv2026

Physics-aware measurement-supervised deep learning enables point spread function inversion in soft X-ray tomography

Soft X-ray tomography (SXT) is an emerging modality for whole-cell 3D imaging in near-native states. However, the effective spatial resolution is limited by optical artifacts characterized by the point spread function (PSF). Standard reconstruction methods force a compromise between structural sharpness and noise, failing to fully resolve these depth-dependent artifacts. By embedding experimentally measured, depth-variant PSFs into a differentiable forward model, we demonstrate a physics-aware computational optimization that bypasses these limitations to recover high-frequency cellular ultrastructure. The structural fidelity was validated using split-tilt Fourier ring correlation (FRC), alongside an experimental bead phantom tomogram, providing supporting evidence that the recovered high-frequency features reflect genuine specimen structure rather than fabricated artifacts. Our method effectively increases FRC spatial resolution and recovers cellular ultrastructure. Furthermore, under sparse-angular subsampling, the framework maintained spatial resolution using half the projection angles, a computational proxy pointing toward the potential for reduced radiation exposure in future acquisitions. This hardware-free, computational approach offers a route toward mitigating the optical and dosimetric constraints that currently limit nanoscale soft X-ray tomography.

Cell Biology↗

bioRxiv2026

Non-enzymatic RNA Glycation is a Metabolic Sensor of Cellular Stress

Non-enzymatic RNA modifications expand the epitranscriptome, encoding a rapid and chemistry-driven response to cellular stress. While methylglyoxal, a reactive glycolytic byproduct of metabolic stress, has been shown to modify proteins and DNA, its impact on RNA has remained unexplored. Here, we identify mRNA as a dynamic substrate of MGO, whose modification is actively regulated by DJ-1 and the glyoxalase detoxification system. We show that mRNA glycation impairs translation and engages both the integrated stress response and the ribotoxic stress pathway, culminating in compromised pancreatic {beta}-cell function and reduced insulin secretion. Notably, this phenotype is alleviated by the frontline antihyperglycemic agent metformin. Together, our findings position mRNA as a direct sensor of metabolic stress and establish RNA glycation as a mechanistic link between glycolytic imbalance, translational stress and disease.

molecular biology↗

bioRxiv2026

Modelling the persistence of post-management disturbance in Calluna vulgaris communities

O_LICalluna vulgaris is often managed in the UK by rotational burning, but this practice has recently been banned on peat with depth greater than 30-40 cm. It is unclear how then to manage the large areas of Calluna on blanket bogs used for sport shooting because without managed burning, fuel loads and wildfire risk will increase as the Calluna ages within the artificially narrow age distributions created by burn management. C_LIO_LIWe developed a model of Calluna mortality and management to understand duration and persistence of post-management effects. This allows us to assess how long it will take to reach a more natural age structure which would allow increased diversity if management ceases. C_LIO_LIOur results show that management effects persist for around 50 years depending on site-specific mortality rates. Active management may therefore be needed either to mitigate the elevated risk of severe wildfire or to speed up this transition. C_LIO_LISome studies have employed, as unmanaged analogues, Calluna stands that were last managed <50 years ago, but such studies may have unintentionally biased their results by observing Calluna still in post-management recovery leading to an over-estimation of wildfire risk associated with more natural blanket bogs. C_LIO_LISynthesis and applications: with the banning of burning as a management tool for Calluna on deep peat, alternative management is now likely needed as our model shows it could take around 50 years for the Calluna to reach a more natural age distribution. Mowing can replicate some of the effects of managed burning but requires repeated intervention and may compress the peat surface from repeated machine tracking. Rewetting and Sphagnum reintroduction may offer a more sustainable management approach to lowering Calluna fuel loads and reducing severe wildfire risk by creating wetter sub-optimal conditions for Calluna growth and thereby altering the competitive balance between Sphagnum and Calluna. Further work is needed to assess the efficacy of rewetting in controlling fuel loads and how this varies with climate and local pressures. More broadly, this work highlights the need to quantify the persistence of past management regimes to understand ecological trajectories. C_LI

ecology↗

bioRxiv2026

Adaptation of white adipocytes to cooler temperatures: impacts on energy metabolism and protein acetylation

Adipocytes throughout the body reside in distinct thermal environments. Visceral adipocytes within the body core are maintained near 37 {degrees}C, whereas those in bone marrow, subcutaneous, and dermal depots occupy cooler regions within the peripheral shell. While brown and beige adipocyte responses to cold stress are well characterized, much less is known about how white adipocytes adapt to moderately reduced temperatures below 37 {degrees}C. Our recent work revealed that cultured adipocytes exposed to 31 {degrees}C, a temperature representative of distal adipose regions, exhibit enhanced mitochondrial function, including increased substrate oxidation and ATP turnover, yet the mechanisms underlying this upregulation remain unclear. Here we show that adaptation to cool temperatures leads to a widespread decrease in protein acetylation in both undifferentiated and differentiated adipocytes, independent of nutrient status, and that this change is readily reversible upon rewarming. Subcellular fractionation and immunoblotting demonstrate that the hypoacetylation coincides with a compartment-specific enrichment of acetylated proteins within mitochondria, indicating selective remodeling of the mitochondrial acetylome. Transcriptomic and biochemical analyses reveal that these temperature-dependent changes occur without alterations in acetyltransferase or deacetylase expression, NAD concentration, or acetyl-CoA availability, suggesting regulation through alternative mechanisms affecting acetyl-CoA flux or enzyme activity. Integrative acetyl-proteomic and metabolomic profiling identifies mitochondrial enzymes, including serine hydroxymethyltransferase 2 (SHMT2) and propionyl-CoA carboxylase (PCCA), whose acetylation correlates closely with changes in associated metabolite pools. Together, these findings establish physiologically relevant cooling as a cell-autonomous regulator of mitochondrial protein acetylation and metabolic adaptation in adipocytes.

cell biology↗

bioRxiv2026

Molecular dynamics of Brodmann Area 22 in development and autism

Challenges in verbal communication are a prominent feature of autism. However, gene regulatory programs in speech-related cortical regions remain poorly characterized. In parallel, it remains unclear whether the heterogeneous genetic factors underlying autism converge on shared neurobiological mechanisms. To address these gaps, we generated paired transcriptomic and epigenomic data from post-mortem human brain tissue across 100 donors. Here, we show that transcriptional differences in the speech-related Brodmann Area 22 in individuals with neurodevelopmental conditions, including autism, are strongest among those with a known genetic diagnosis. A similar but attenuated signature is observed in those without a genetic diagnosis. These transcriptional differences are most pronounced in neurons, with glutamatergic L4/5 intratelencephalic neurons affected across multiple modalities. Finally, multimodal analysis implicates altered RFX3-dependent networks as a central hub in autism, particularly among L4/5 intratelencephalic neurons in non-verbal individuals. Together, our study identifies regulatory architecture linking chromatin state, transcriptional output, and variation in verbal ability in autism.

neuroscience↗

bioRxiv2026

ATF4 Coordinates Transcriptomic and Structural Adaptations in Aging Muscle

Aging is associated with a progressive loss of skeletal muscle function, known as sarcopenia; however, the molecular mechanisms coordinating cellular stress responses and structural adaptations permissive of sarcopenia remain incompletely understood. In our previous studies, we found aging differentially impacted mitochondrial networks by muscle, suggesting unique stress thresholds and response activation. Here, we investigate the role of activating transcription factor 4 (ATF4), a master regulator of the integrated stress response (ISR), in aged quadriceps muscle using complementary patient and aging mouse models. Older adults exhibited a marked decrease in aerobic capacity, muscle strength, and endurance when compared with young participants. These results paralleled findings in aged mice, with significant loss of muscle mass across multiple hindlimb muscles. Ultrastructural analysis revealed substantial age-related changes in mitochondrial morphology, including increased volume, surface area, and branching index, as well as a shift toward larger, more complex mitochondria. Our data indicate that ATF4 binds directly to the promoter region of the gene encoding TFAM, suggesting a transcriptional regulatory relationship to support DNA stability. These structural and transcriptional changes likely impair oxidative capacity and drive a feed-forward cycle of mitochondrial dysfunction and ISR activation. Our findings indicate that ATF4 coordinates transcriptomic and structural adaptations in aging muscle, identifying the ISR pathway as a potential therapeutic target for preserving muscle function in older adults.

cell biology↗

bioRxiv2026

Structural Basis for Dual Peptidoglycan Hydrolysis by an E. faecium Minhovirus Tail Spike Lysin

Bacteriophages rely on breaching the bacterial cell wall as the first step of infection. We characterise ORF11, a putative tail-spike lysin from the 19 kbp Minhovirus SHEF14, a podovirus infecting Enterococcus faecium. Bioinformatic analyses indicate that ORF11 comprises four domains: a predicted glycosyl hydrolase (D1) a cysteine, histidine-dependent amidohydrolases/peptidases (CHAP, D4), separated by a helical linker (D2) and a CHAP-like domain (D3). This modular organisation is conserved among Enterococcus minhoviruses but differs markedly from analogous proteins in Copernicusvirus phage and related staphylococcal phage. ORF11 2.1 [A] crystal structure reveals an unusual dimeric assembly. The predicted glycosyl hydrolase and CHAP peptidase domains occupy opposite ends of the protein, bridged by the two other domains positioned at the dimer interface. Biochemical assays using recombinant ORF11 and LC-MS/MS confirmed dual peptidoglycan-degrading activity. ORF11 functions as both an N-acetylglucosaminidase and a D, D-endopeptidase, cleaving the bond between the D-alanine in position-4 and the D-aspartate residue at the end of the side chain. Together, these results provide the first structural description of a podovirus tail-spike lysin and demonstrate its bifunctional enzymatic activity. This dual action likely facilitates initial surface recognition and localised peptidoglycan degradation during infection of E. faecium, offering new insights into how minimal-genome phage target this clinically significant antimicrobial-resistant pathogen.

biochemistry↗

bioRxiv2026

Amoxicillin induces gut dysbiosis leading to long term suppression of type-17 immune tone in the lungs

T-helper (Th)-17 lymphocytes are central mediators of adaptive type 17 immunity. Decreased type-17 signaling increases severity of infections in humans and mice. However, detrimental effects of excessive type 17 responses in autoimmune and other inflammatory diseases highlight a need for type-17 immune calibration to support beneficial host defense requirements. Mechanisms of type 17 calibration are poorly understood. A gut-lung axis has been proposed to coordinate homeostatic protection and acute host defense. Factors that acutely alter the gut microbiome are heterogeneous and include acute intestinal infections, non-infectious colitis, and medical treatments such as antibiotics. How changes in the gut microbiome affect lung immune tone during homeostasis and acute pulmonary infections are also poorly understood. Prior studies have shown that antibiotics reduce expression of IL-17-mediated host defense in the gut. Since gut microbial homeostasis influences Th17 cell numbers in both the intestine and remote tissues, we postulated that antibiotic treatment would result in gut dysbiosis and weakened type-17 host defense in the lungs. We found that amoxicillin induces significant dysbiosis that is long-lasting and that there is a long-term decrease in type-17 tone in the lungs. We also found that in mice lacking the gut mucin, Muc2, Th17 cells increased in the lungs following inflammatory challenge. These findings suggest that antibiotic-induced dysbiosis can decrease lung immune defenses for long periods of time after cessation of antibiotic treatment.

immunology↗

bioRxiv2026

Metaproteomics Reveals Functional Rewiring and Metabolic Limits in Synthetic Bacterial Consortia Degrading Complex Waste Oils

Elucidating the metabolic mechanisms underpinning complex hydrocarbon biodegradation by microbial consortia benefits from integration of community-level genomic and proteomic data. Here, we deployed a combined metagenomics and metaproteomic analysis to characterize functional dynamics within bacterial consortia enriched from hydrocarbon-contaminated soil, using military waste oil as the sole carbon source. Metagenomic profiling revealed a consortium dominated by Stenotrophomonas maltophilia alongside established hydrocarbon-degrading taxa including Pseudomonas fluorescens, Pseudomonas putida, and Delftia acidovorans. Quantitative label-free metaproteomics uncovered dynamic taxonomic restructuring and metabolic reorientation, with Delftia and Achromobacter proteins enriched during active degradation, indicating niche advantages under chronic contamination. Enzymatic mapping resolved expression of degradation pathways targeting aliphatic and aromatic substrates, with one specific consortium, BPS4, exhibiting preferential catabolism of catechol intermediates channeled toward the tricarboxylic acid cycle, supported by elevated protein abundance of oxidoreductases, dehydrogenases, and {beta}-oxidation enzymes. Gene ontology enrichment revealed heightened nitrogen cycling capacity, including upregulation of nitric oxide biosynthesis and nitrate reductase, suggesting adaptive nitrogen metabolism coupled to hydrocarbon mineralization. In-vitro batch-fed adaptation studies demonstrated limited improvements in degradation of recalcitrant fractions, particularly branched aliphatics and organophosphates, revealing intrinsic metabolic constraints within the consortium. These findings underscore the complementary utility of metaproteomic interrogation for resolving both population-level dynamics and functional metabolic capabilities, while highlighting fundamental limitations in expanding the degradative spectrum of enriched consortia through conventional adaptation strategies. This work establishes a foundation for rational ecological engineering of hydrocarbon-remediating consortia informed by metaproteomics. ImportanceWaste oils from transport and industry are complex and toxic mixtures that are difficult and expensive to treat using conventional chemical methods. Microorganisms offer a sustainable alternative, but their performance depends on how different species work together and where their metabolic limits lie. In this study, we show how advanced protein-based analysis using quantitative metaproteomics, can reveal microbial community activities whilst breaking down real-world waste oils. We enriched bacteria from contaminated soils and then identified the enzymes driving effective degradation. We investigated how communities reorganise under chemical stress, and why certain oil components remain resistant even after long-term adaptation. By uncovering both strengths and limitations of naturally enriched microbial consortia, this work provides practical insights for designing more reliable biological treatments for oil-based wastes and even contaminated environments, supporting greener approaches to pollution management and industrial waste recycling.

microbiology↗

bioRxiv2025

The role of MICOS in modulating mitochondrial dynamics and structural changes in vulnerable regions of Alzheimer's Disease

Mitochondrial contact site and cristae organizing system (MICOS) complexes are critical for maintaining the mitochondrial architecture, cristae integrity, and organelle communication in neurons. MICOS disruption has been implicated in neurodegenerative disorders, including Alzheimers disease (AD), yet the spatiotemporal dynamics of MICOS-associated neuronal alterations during aging remain unclear. Using three-dimensional reconstructions of hypothalamic and cortical neurons, we observed age-dependent fragmentation of mitochondrial cristae, reduced intermitochondrial connectivity, and compartment-specific changes in mitochondrial size and morphology. Notably, these structural deficits were most pronounced in neurons vulnerable to AD-related pathology, suggesting a mechanistic link between MICOS disruption and the early mitochondrial dysfunction observed in patients with AD. Our findings indicate that the loss of MICOS integrity is a progressive feature of neuronal aging, contributing to impaired bioenergetics and reduced resilience to metabolic stress and potentially facilitating neurodegenerative processes. MICOS disruption reduced neuronal firing and synaptic responsiveness, with miclxin treatment decreasing mitochondrial connectivity and inducing cristae disorganization. These changes link MICOS structural deficits directly to impaired neuronal excitability, highlighting vulnerability to AD-related neurodegeneration. These results underscore the importance of MICOS as a critical determinant of neuronal mitochondrial health and as a potential target for interventions aimed at mitigating AD-related mitochondrial dysfunction.

neuroscience↗

bioRxiv2025

Disruption of Cell-Type-Specific Molecular Programs of Medium Spiny Neurons in Autism

Autism spectrum disorders (ASD) are highly heritable neurodevelopmental conditions with major contributions from rare genetic variants. Most studies have focused on cortical mechanisms; even growing evidence implicates subcortical circuits in ASD etiology. To systematically map developmental and molecular alterations beyond the cortex, we profiled lineage relationships across five brain regions in an ASD mouse model. Most prominent changes emerged in the striatum, a hub for learning and motor control. Furthermore, we performed single-nucleus multiomic profiling of human putamen from ASD and neurotypical donors revealed cell-type-specific transcriptomic and regulatory alterations. Differential expression converged on synaptic and energy metabolic dysfunctions in D1 striosome medium spiny neurons (MSNs), coupled with astrocytic remodeling of synaptic support. Gene regulatory network analysis identified EGR3 and EGR1 as key transcriptional regulators of ASD-associated programs of D1 MSNs. Together, these results establish the striatum as a central node of ASD convergence and provide a multiomic resource for dissecting its subcortical mechanisms.

neuroscience↗

bioRxiv2025

Integrative multi-omics uncovers skeletal muscle enhancer programming of cardiorespiratory fitness

Cardiorespiratory fitness (CRF) is a heritable trait associated with improved metabolic health and longevity. To identify regulatory mechanisms underlying CRF, we integrated 546 transcriptomic and epigenomic profiles from skeletal muscle of 128 genetically heterogeneous rats selectively bred for high and low running capacity, a model that mirrors CRF-associated traits in humans. Selection drove genetic convergence in coordinated skeletal muscle enhancer networks linked to lipid metabolism and angiogenesis genes. We validated thousands of these genetic effects through integration of 426 genotype, gene expression, and chromatin accessibility profiles in an independent HCRxLCR F2 population (n=147). These 972 multi-omics profiles show that CRF-associated genetic variation reshapes the chromatin landscape to support energy metabolism and oxygen delivery, offering a molecular framework for identifying targets to reduce cardiometabolic disease risk. Glossary of TermsO_LIModality/modalities: essentially a dataset. A modality can refer to one or multiple of ATAC-Seq, H3K27ac & additional histone modifications, chromatin states such as Strong Enhancer, and/or RNA-Seq C_LIO_LIPeaks: only refers to ATAC-Seq and CUT&Tag data C_LIO_LIFeatures: can collectively refer to genomic regions of one or more modality classes (genes, peaks, and/or chromatin state regions) C_LIO_LIVariant(s): SNP(s) C_LI O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/679855v2_ufig1.gif" ALT="Figure 1"> View larger version (56K): org.highwire.dtl.DTLVardef@1ee52eorg.highwire.dtl.DTLVardef@5848b7org.highwire.dtl.DTLVardef@debaforg.highwire.dtl.DTLVardef@1ec43ba_HPS_FORMAT_FIGEXP M_FIG C_FIG

genomics↗

bioRxiv2024

Failed Double-Blind Replication of Offline 5Hz-rTUS-Induced Corticospinal Excitability

IntroductionTranscranial ultrasound stimulation (TUS) is a promising new form of non-invasive neuromodulation. As a nascent technique, replication of its effects on brain function is important. Of particular interest is offline 5Hz repetitive TUS (5Hz-rTUS), originally reported by Zeng and colleagues [1] to elicit lasting corticospinal excitability increases, with large effect sizes. Material and methodHere, we conducted a pre-registered (https://osf.io/p5n4q) replication of this protocol that benefitted from three additional features: double-blind application of TUS, neuronavigation for consistent TMS positioning, and acoustic simulations to assess M1 target exposure to TUS. Changes in resting motor thresholds (rMT), motor-evoked potential (MEP) amplitude, short-interval intracortical inhibition (SICI) and intracortical facilitation (ICF) in response to TUS (5Hz-rTUS vs. sham) were measured in the right first dorsal interosseous (FDI), abductor digiti minimi (ADM) and abductor pollicis brevis (APB) muscles, with unbiased selection of participants. Transducer location was determined by the TMS-hotspot for motor representations of the right FDI, as in the original work. ResultsNo significant effects of 5Hz-TUS (vs sham) were observed. Post-hoc simulations showed considerable variability of the acoustic focus, which was outside the anatomical M1-hand area in 67% of participants - in line with the known poor correspondence of TMS-hotspot location and M1-hand area. ConclusionOur results indicate that the effect sizes of the neuromodulatory effects of 5Hz-rTUS on M1 may be more variable than previously appreciated. We suggest that double-blinding, neuronavigated TMS, individualised acoustic simulations for TUS targeting and pre-registration will aid reproducibility across studies.

neuroscience↗

bioRxiv2024

Reciprocal links between methionine metabolism, DNA repair and therapy resistance in glioblastoma

Glioblastoma (GBM) is uniformly lethal due to profound treatment resistance. Altered cellular metabolism is a key mediator of GBM treatment resistance. Uptake of the essential sulfur-containing amino acid methionine is drastically elevated in GBMs compared to normal cells, however, it is not known how this methionine is utilized or whether it relates to GBM treatment resistance. Here, we find that radiation acutely increases the levels of methionine-related metabolites in a variety of treatment-resistant GBM models. Stable isotope tracing studies further revealed that radiation acutely activates methionine to S-adenosyl methionine (SAM) conversion through an active signaling event mediated by the kinases of the DNA damage response. In vivo tumor SAM synthesis increases after radiation, while normal brain SAM production remains unchanged, indicating a tumor- specific metabolic alteration to radiation. Pharmacological and dietary strategies to block methionine to SAM conversion slowed DNA damage response and increased cell death following radiation in vitro. Mechanistically, these effects are due to depletion of DNA repair proteins and are reversed by SAM supplementation. These effects are selective to GBMs lacking the methionine salvage enzyme methylthioadenosine phosphorylase. Pharmacological inhibition of SAM synthesis hindered tumor growth in flank and orthotopic in vivo GBM models when combined with radiation. By contrast, methionine depletion does not reduce tumor SAM levels and fails to radiosensitize intracranial models, indicating depleting SAM, as opposed to simply lowering methionine, is critical for hindering tumor growth in intracranial models of GBM. These results highlight a new signaling link between DNA damage and SAM synthesis and define the metabolic fates of methionine in GBM in vivo. Inhibiting radiation-induced SAM synthesis slows DNA repair and augments radiation efficacy in GBM. Using MAT2A inhibitors to deplete SAM may selectively overcome treatment resistance in GBMs with defective methionine salvage while sparing normal brain.

cancer biology↗