Biology subjects

Miller, R.

Publications and source records attributed to Miller, R..

At least 19 recordsLinked to original sources

bioRxiv2026

Intrastriatal injection of amplified Lewy body dementia (LBD) fibrils in mice results in accumulation of fibrils with LBD structure and neurodegeneration

The defining neuropathologic feature of Lewy body dementia (LBD) and Parkinson's disease (PD) is alpha-synuclein (Asyn) fibril accumulation in Lewy bodies (LBs) and Lewy neurites (LNs). Recently, we developed a method to amplify Asyn fibrils derived from human post-mortem LBD brain tissue and determined that the structure of amplified Asyn fibrils is highly similar to the structure of fibrils directly isolated from LBD brain tissue. We examined whether injection of these fibrils into humanized Asyn mice recapitulates pathologic Asyn fibril accumulation with human LBD Asyn fibril structure. We utilized mice containing a null allele in the mouse Snca gene and a human SNCA transgene. Intrastriatal injection of amplified Asyn fibrils derived from 3 different LBD autopsy cases resulted in cytoplasmic and neuritic phospho-Asyn accumulation in the striatum, cortex, and amygdala bilaterally at 3mpi. Fibril injections produced a reduction in the number of dopaminergic neurons in the substantia nigra, with LB-like inclusions in surviving neurons, and produced impaired sensorimotor function as measured on the accelerating rotarod test. Fibril accumulation increased 3.6-fold between 1 and 3 months post-injection (mpi) and was relatively stable between 3 and 5 mpi. We amplified fibrils from mouse brain tissue 5 mpi and collected single-particle cryo-EM data. High resolution 3D structural information could not be obtained because of the lack of helical twist, but comparison of the cryo-EM 2D class averages with those derived from amplified fibrils prepared from human LBD tissue revealed multiple matching classes, indicating that the Lewy fold is maintained in the fibrils that accumulate in mouse brain tissue. Our findings demonstrate an approach to model the accumulation of Asyn fibrils with LBD structure in mice, providing a platform for investigating disease mechanisms, imaging biomarkers, and therapeutics targeting Asyn in PD/LBD.

neuroscience↗

bioRxiv2026

Multi-Tissue Metabolomic Signatures of Five Longevity Interventions Converge on Ergothioneine and Lipid Remodeling in Male UM-HET3 Mice

The pace of aging can be delayed by mutations, dietary manipulations, and drugs, yet the metabolic mechanisms underlying longevity interventions remain poorly understood. Here we present a multi-tissue metabolomic analysis of male UM-HET3 mice treated from 4 to 12 months of age with five validated longevity interventions: rapamycin, acarbose, 17-estradiol, canagliflozin, or caloric restriction. Using a feature-stabilized XGBoost pipeline applied to seven tissues, we show that metabolomic profiles can identify treated mice as likely recipients of a lifespan-extending intervention well before survival differences emerge. A leave-one-intervention-out procedure confirmed that models trained on any four interventions successfully classified mice from a fifth, unseen intervention, implying shared metabolic alterations across mechanistically distinct treatments. The most influential metabolites -- defined as the minimum set explaining 50% of cumulative model gain -- differed substantially across tissues. Only ergothioneine, a dietary antioxidant, ranked highly in more than two tissues: it was elevated by all five interventions in plasma and brain, and by four of five in muscle. Enrichment analyses further identified coordinated remodeling of lipid classes in plasma, perigonadal fat, and kidney. These findings reveal tissue-specific metabolic reprogramming shared across mechanistically distinct longevity interventions and, pending validation against interventions that do not extend lifespan, suggest a path toward metabolomic screening of candidate anti-aging drugs.

Molecular Biology↗

bioRxiv2026

Geroprotective interventions preserve trabecular bone during ageing in female mice

Geroprotectors extend lifespan and improve several aspects of healthspan, yet their effects on skeletal ageing remain poorly understood. They hold potential advantages over current bone-targeted osteoporosis therapies, as they may simultaneously improve bone, neuromuscular function, and vision, thereby reducing the risk of falls, the major cause of fractures. Here we examined, for the first time, the long-term effects of rapamycin, acarbose, and 17-estradiol, administered at lifespan-extending doses on trabecular and cortical bone architecture in male and female UM-HET3 mice measured with micro-computed tomography at 12 and 22 months of age. Bayesian modelling analysis reveals that all interventions produced responses in trabecular bone in females at 22 months. These effects were driven mainly by increases in trabecular number, with little evidence for changes in trabecular thickness. In contrast, treatment effects in males were generally negligible. Cortical responses were modest. Moderate increases in cortical area fraction were observed in females treated with rapamycin or 17-estradiol at 22 months, whereas cortical thickness remained largely unchanged, suggesting a geometrical rather than anabolic effect. Interestingly, geroprotectors strongest skeletal responses in females contrasts with the predominantly male-biased lifespan extension reported for acarbose and 17-estradiol, suggesting differential mechanisms mediating lifespan extension and bone structure preservation.

pharmacology and toxicology↗

bioRxiv2026

Large-scale brain dynamics are organized by a directional coordination hierarchy

The cortical hierarchy is a central organizing feature of brain structure and function, yet whether it is continuously expressed in spontaneous neural activity remains unresolved. Here, we show that resting-state brain dynamics are organized along a directional coordination axis that recapitulates hierarchical information flow. Focusing on the directional dynamics of interregional coordination, we identify three recurrent coordination regimes: a feedback-dominated mode in which transmodal cortex leads sensory systems, a feedforward-dominated mode reflecting the reverse, and an integrative mode of balanced bidirectional exchange. These modes define a low-dimensional coordination landscape that replicates across four independent cohorts, establishing directional structure as a stable property of the adult brain. In schizophrenia, this landscape is selectively reorganized: feedback-dominated coordination becomes less persistent, integrative coordination becomes more persistent, and global dynamics shift toward faster convergence and reduced entropy, reflecting a loss of directional constraint and dynamical flexibility. These alterations track symptom severity, cognitive performance, and medication exposure. Mediation analyses suggest that medication-related reductions in integrative coordination are statistically routed through feedforward dynamics, consistent with dopaminergic modulation of recurrent cortical loops as a candidate mechanism. Together, these findings identify directional functional dynamics as a fundamental and clinically informative axis of large-scale brain organization.

neuroscience↗

bioRxiv2026

The effect of long-term management on wild pig (Sus scrofa x domesticus) populations across the southeastern United States

O_LIThe management of invasive species often emphasizes removals to manage populations. However, evaluating the success of this management technique remains challenging, especially at large scales. Understanding the relationship between removal intensity and population growth is essential for determining when management achieves desired outcomes. C_LIO_LIWe used management removal data (removal resources [e.g. trapping] and relative effort [trap nights]) to estimate population density, demographic structure, and growth rates of invasive wild pigs (Sus scrofax domesticus) across a large landscape. From the management data and population estimates, we inferred population trajectories in the absence of removals and quantified the proportion of the population removed by the most widely used methods to control wild pigs. We then compared observed removal intensities and population growth rates to predict expected population trajectories immediately after management occurs. C_LIO_LIResults suggest substantial spatial and temporal variation in wild pig growth rates and variation in the effectiveness of removal efforts. Additionally, removing wild pigs at higher densities had a greater effect on limiting population growth than removals conducted at lower densities, though both are important. However, on large properties, removal intensity was often insufficient to offset population growth, indicating that management effort does not scale to large areas. C_LIO_LIThese results demonstrate how removal data and population modeling can provide robust inference on population dynamics and management effectiveness, offering a scalable framework for evaluating and improving invasive species control programs. We also discuss the current limitation of how effort is defined for different large-mammal removal techniques, and offer potential solutions for a more complete definition, such as going beyond trap nights and including constraints on personnel, equipment, and logistics. C_LI

ecology↗

bioRxiv2026

A new mRNA antigen vaccine induces potent B and T cell responses and in vivo protection against SARS-CoV-2

The SARS-CoV-2 mRNA vaccine provides effective protection against viral infection and severe disease by inducing efficient adaptive immunity. However, vaccine efficacy is decreased against emerging variants, and immune memory is relatively short-lived. Here, we added new T cell epitopes to the RBD (receptor-binding domain) mRNA vaccine and identified a SARS-CoV-2 membrane epitope that significantly improved vaccine-induced immunity and protection in vivo. That new vaccine, designated G1-C, induced 8.2-fold higher levels of RBD-specific antibodies than did RBD and enhanced spike-specific T cell and B cell responses. Remarkably, the G1-C modulated hematopoietic stem cell (HSC) differentiation and increased levels of B and NK cells by regulating multiple signaling pathways in bone marrow potentially via Fos, Klf4, and Klf6 transcription factors. Altogether, these findings identify a new vaccine candidate to control viral infection by affecting the lymphoid-myeloid lineage bias and suggest the potential role of T cell epitopes in vaccine design and development.

immunology↗

bioRxiv2026

Development of cognition in corvids

Corvids - members of the crow family - exhibit some of the most sophisticated cognitive abilities outside the primate lineage, yet the developmental origins of many of these abilities remain poorly understood. Here, we present a systematic review of the past 20 years (from 2005) of empirical research on corvid cognitive development, synthesising evidence across core/ foundational, social and physical cognitive domains. Using a structured search strategy and detailed coding framework, we identified 47 relevant studies spanning 16 corvid species. We evaluate and discuss developmental trajectories, species/ taxa-level variation and methodological robustness across studies. For within and between-taxa comparisons, we particularly focus on the best represented abilities in the coded sample: 1) object permanence and caching; 2) tool-use/ manufacture; 3) object manipulation and play; and 4) gaze following. Corvid developmental patterns show both parallels and divergences from those documented in primates and other taxa. However, the existing corvid evidence base is constrained by small samples, captive biases, limited longitudinal data and under-representation of key cognitive abilities, such as executive function, causal reasoning, self-control, metacognition, spatial memory and social learning. We outline critical gaps and future directions, emphasising the need for comparative, longitudinal and ecologically grounded approaches, including the science of magic and Theory of Mind, to better understand how early-life cognition shapes later behaviour, cognition and fitness in this model avian family.

animal behavior and cognition↗

bioRxiv2025

The Nicotinamide Salvage Pathway is a Metabolic Vulnerability of High-Risk MDS Stem Cells

High-risk myelodysplastic syndrome (HR-MDS) is a malignant clonal disorder originating in hematopoietic stem and progenitor cells (HSPCs). The current standard of care for HR-MDS patients is hypomethylating agents; however, the response rate is poor. There is thus a need to explore vulnerabilities of HR-MDS HSPCs for better clinical outcomes. We demonstrate that HR-MDS HSPCs have significant upregulation of metabolic proteins required for glycolysis, citric acid cycle, and oxidative phosphorylation. Consistently, we see increased oxygen consumption rate in HR-MDS HSPCs compared to healthy, suggesting an increased metabolic rate. Corroboratively, compared to healthy HSPCs, HR-MDS HSPCs have increased abundance of mitochondrial complex I proteins, which are NADH dehydrogenases, and crucial for energy production. Therefore, we investigated whether HR-MDS HSPCs are functionally reliant on NAMPT, the rate-limiting enzyme in the nicotinamide salvage pathway of NAD anabolism. NAMPT inhibition significantly decreased NAD(H) in HR-MDS HSPCs. Consequently, NAMPT inhibition reduced the oxygen-consuming capacity of HR-MDS-HSPCs compared to healthy. Importantly, NAMPT inhibition significantly impaired the self-renewal and colony-forming potential, increased cell death and reduced disease burden specifically of HR-MDS HSPCs, compared to healthy controls. Collectively, our data suggest that NAMPT is selectively required for the function and survival of HR-MDS HSPCs representing a promising therapeutic target.

cancer biology↗

bioRxiv2025

spaTransfer: transfer learning for single-cell and spatialtranscriptomics data using non-negative matrix factorization

Recent advances in spatially-resolved transcriptomics have enabled profiling of gene expression in a spatial context, which has led to the generation of large-scale single-cell and spatial atlases with computationally-derived cell type or spatial domain labels. An increasingly important task with these data has become the transfer of cell type or spatial domain annotations from a given reference (or source) atlas into a new target tissue or sample. The reference and target datasets could be at different resolutions or measured on different experimental platforms. Here, we present a method to perform cross-platform transfer learning that takes as input single-cell or spatial domain labels from a reference atlas or dataset and transfers the labels to a target dataset at a similar or different resolution. Specifically, we use non-negative matrix factorization (NMF) on the reference data to identify factors associated with labels of interest and project these factors into the target dataset to label each new observation. We use a multinomial model with the factors as covariates and labels as the response to predict labels in the target dataset. In contrast to existing approaches, the advantage of our approach is interpretability, without compromising on accuracy. We demonstrate the performance of the method in two human brain tissues and show that our model identifies spatially coherent domains in the target datasets with concordance of marker gene expression. We implement spaTransfer in open-source software as an R package (github.com/cindyfang70/spaTransfer).

bioinformatics↗

bioRxiv2025

Improving a removal model for evaluating density changes in a widespread invasive species

Invasive species pose a significant threat to native ecosystems. Effective management of invasive species requires accurate abundance estimates through time, particularly at large spatial scales. Traditional methods of abundance estimation from management data, such as camera trap studies and harvest data, face challenges due to variations in sampling protocols and effort standardization. We propose an extension to a previously calibrated removal model by integrating into an Integrated Population Model (IPM) and incorporating multiple removal techniques. The model utilizes management and land cover data, both of which are readily available in many invasive species systems. We evaluate the model with a simulation study, then apply it to management data on wild pigs (Sus scrofa). Simulations showed that the model performs well across a wide range of densities and population trajectories. Density estimates were the most biased when effort and density were low and sharpshooting or fixed wing aircraft were used to conduct removals. We discuss the advantages and limitations of using removal models for invasive species management, how the model performs with respect to previous removal modeling efforts, how performance might be improved through changes to data management practices, and the models potential to inform decision-making processes for invasive species management. Data availabilityThe data for this manuscript is available from Dryad (http://datadryad.org/share/gFz3ClanB-8yPB5lQ1x-dl1JqI07CP3SX92P-9UyfaU). The code for this manuscript will is available from Zenodo (10.5281/zenodo.15784366).

ecology↗

bioRxiv2025

Insights from farming Macrocystis pyrifera offshore: phenotypic analysis, genome-wide association studies, genomic selection

Seaweed farming, as a part of aquaculture, offers a sustainable alternative to modern agricultural practices; however, genetic enhancement and breeding programs for most species are underdeveloped. We aimed to advance seaweed domestication by focusing on giant kelp (Macrocystis pyrifera), the fastest-growing haplodiplontic brown alga, which has significant ecological and commercial importance. We analyzed phenotypic data from two offshore experimental farms conducted in 2019 and 2020, which involved hundreds of outplanted genetically diverse sporophytes. We found that outplanting season and farm design had significant effects on giant kelp biomass. Broad-sense heritability estimates showed moderate (0.27-0.50) genetic contributions to two phenotypes, carbon content and total biomass. Genome-wide association studies for these phenotypes resulted in three statistically significant SNPs, located near or within genes involved in carbohydrate metabolism and cytoskeletal functions. In addition, we applied genomic selection models that integrated sporophyte phenotypes and parental gametophyte genotypes. These models utilized reduced sets of GWAS-ranked SNPs obtained by a procedure based on linkage disequilibrium estimations. Model testing yielded cross-validation accuracy values of up to 0.84 and predictive accuracy values of up to 0.40, demonstrating the potential of marker-assisted breeding for phenotype improvement. Our results provide foundational genomic resources and tools for domesticating and breeding M. pyrifera, offering a basis for developing giant kelp varieties with desirable traits.

genomics↗

bioRxiv2025

Blocking somatic repeat expansion and lowering huntingtin via RNA interference synergize to prevent Huntingtons disease pathogenesis in mice

Huntingtons disease (HD) is a progressive neurodegenerative disorder with no approved therapies. Two major molecular drivers--somatic expansion of inherited CAG repeats and toxic mutant HTT (mHTT) variants--lead to neuronal dysfunction. Despite multiple trials, HTT-lowering strategies have not shown meaningful clinical benefit. Using therapeutic divalent siRNAs, we assessed the long-term impact of silencing MSH3 (a key regulator of somatic expansion), HTT, or both. In Q111 HD mice (>110 CAGs), which exhibit robust expansion, mHTT inclusions, and transcriptional dysregulation by 12 months, long-term MSH3 silencing blocked expansion, reduced inclusions, and reversed gene expression changes. HTT silencing alone had limited effect, but combined MSH3/HTT targeting synergistically eliminated inclusions and restored transcriptomic profiles. Parallel treatment in wild-type mice showed no toxicity, supporting the safety of long-term intervention. These findings position somatic expansion as a promising therapeutic target and demonstrate the potential of RNAi-based co-silencing of MSH3 and HTT as a disease-modifying strategy for HD.

neuroscience↗

bioRxiv2025

Stress induces distinct social behavior states encoded by the ventral hippocampus

A single, acute traumatic experience can result in a host of negative impacts on behavior, such as increased violence, reduced sociability, and exaggerated fear responses. Despite the large body of research on the neurobiology of stress, we have a poor understanding of how trauma rewires social circuits in the brain. To identify how social circuits are re-organized by stress, we interrogated the role of the ventral hippocampus (VH), a key node for both the orchestration of emotionally-relevant behavior and the integration of sensory information. Using a footshock-based model of traumatic stress, we established that a single exposure to a series of unpredictable, inescapable footshocks was sufficient to negatively alter social behavior, resulting in increased violence and social hesitancy. Critically, we found that stress-induced changes to social behavior engaged neural ensembles in the VH, a critical node in the regulation of emotion states. Using a virally-mediated, activity-dependent cellular tagging approach to label two neural ensembles activated by temporally distinct experiences, we were surprised to find that stress-induced violence and stress-induced social hesitancy recruited largely non-overlapping populations of cells in the VH, in contrast to higher degrees of ensemble overlap in unstressed control mice, suggesting that stress biases the brain towards stronger differentiation of distinct social states. Additionally, we found that activity of VH neurons was required for stress-induced aggression. Finally, we found that stress-induced changes to social behaviors and VH activity profiles could be reversed by the introduction of social buffering post-stress. Collectively, our findings suggest that stress drives the VH to dissociably encode specific behavioral states, rather than a single state of negative valence, consistent with a role for multiple structures and distributed ensembles in the modulation of traumatic stress.

neuroscience↗

bioRxiv2025

DNA-based delivery of incretin receptor agonists using MYO Technology leads to durable weight loss in a diet-induced obesity model

Therapeutic proteins have seen a substantial increase in clinical development and use across many disease areas. Despite their broad applicability, significant drawbacks limit access to many of these drugs, including: i) high manufacturing costs; ii) administration via time-consuming infusions; iii) frequent dosing, sometimes even daily; and iv) requirements for low temperature storage. MYO Technology was developed to overcome these barriers. The MYO Technology platform consists of therapeutic-encoding plasmid DNA (pDNA), and a proprietary medical device for intramuscular injection and delivery of electrical pulses. These pulses enable the in vivo electroporation of muscle cells and uptake of injected pDNA, leading to the production, secretion, and delivery of the therapeutic protein into peripheral circulation. MYO Technology offers several advantages over standard delivery of therapeutic proteins; pDNA manufacturing is a simpler and less specialized process compared to protein manufacturing, and pDNA is very stable and lacks most cold chain requirements. Furthermore, administration using MYO Technology takes only a few minutes, and the serum level of a therapeutic protein can potentially be maintained for many months without the need for redosing. Incretin receptor agonists (IRAs) are a class of therapeutic proteins that have recently come to prominence as powerful weight and glucose control drugs, and are used for the treatment of type 2 diabetes (T2D) and obesity. Semaglutide and tirzepatide, currently the most widely used within this class, are both potent molecules, but have a short half-life, requiring weekly administration by subcutaneous injections. Moreover, since their clinical benefits rapidly disappear upon treatment cessation, T2D and obese patients may have a life-long dependency on IRAs, and the requirement for weekly injections can negatively affect the quality of life and the adherence to therapy, as well as create a significant financial burden. Therefore, increasing the interval between injections has become one of the major goals in the field. Here, we present our preclinical studies on the delivery of IRAs with MYO Technology. Animal proof-of-concept studies demonstrate that MYO Technology-delivered IRAs are functional, and efficacious in promoting long-lasting weight and glucose control in mouse models of diet-induced obesity. Moreover, engineering the IRAs to facilitate blood-brain barrier penetration further enhances treatment efficacy, with benefits persisting beyond one year following a single administration. Together, these findings highlight MYO Technologys potential to transform care for patients with T2D and obesity by enabling long-lasting therapeutic effects with minimal dosing, ultimately improving quality of life and treatment adherence.

physiology↗

bioRxiv2025

Therapy resistance in AML is mediated by cytoplasmic sequestration of the transcriptional repressor IRF2BP2

While the development of venetoclax with azacitidine (ven/aza) has improved AML therapy, drug resistance remains a major challenge. Notably, primary ven/aza-resistant AML are frequently reliant on MCL1, however, the underlying mechanisms remain unclear. Co-immunoprecipitation of MCL1 from ven/aza-resistant AML samples coupled with mass spectrometry analysis identified the transcriptional repressor Interferon Regulatory Factor 2 Binding Protein 2 (IRF2BP2) as an MCL1 binding partner. This interaction results in cytoplasmic IRF2BP2 localization and loss of transcriptional repression within ven/aza-resistant leukemic stem cells (LSC). Consequently, ven/aza-resistant LSC have increased IRF2BP2 target gene expression, including acyl-CoA synthetase long-chain family member 1 (ACSL1), an essential rate-limiting enzyme for fatty acid oxidation (FAO). Inhibition of ACSL1 functionally impaired ven/aza-resistant LSC through a depletion of long-chain acyl-carnitine metabolites and FAO. Collectively, these data provide evidence for a previously undescribed mechanism by which MCL1 mediates IRF2BP2 cytoplasmic sequestration and consequent de-repression of ACSL1, thereby promoting ven/aza-resistance in AML.

cancer biology↗

bioRxiv2025

Long-term hunter-gatherer continuity in the Rhine-Meuse region was disrupted by local formation of expansive Bell Beaker groups

The first phase of the ancient DNA revolution painted a broad-brush picture of European Holocene prehistory, whereby 6500-4000 BCE, farmers descending from western Anatolians mixed with local hunter-gatherers resulting in 70-100% ancestry turnover, then 3000-2500 BCE people associated with the Corded Ware complex spread steppe ancestry into north-central Europe. We document an exception to this pattern in the wider Rhine-Meuse area in communities in the wetlands, riverine areas, and coastal areas of the western and central Netherlands, Belgium and western Germany, where we assembled genome-wide data for 109 people 8500-1700 BCE. Here, a distinctive population with high hunter-gatherer ancestry ([~]50%) persisted up to three thousand years later than in continental European regions, reflecting limited incorporation of females of Early European Farmer ancestry into local communities. In the western Netherlands, the arrival of the Corded Ware complex was also exceptional: lowland individuals from settlements adopting Corded Ware pottery had hardly any steppe ancestry, despite a characteristic early Corded Ware Y-chromosome. The limited influx may reflect the unique ecology of the regions river-dominated landscapes, which were not amenable to wholesale adoption of the early Neolithic type of farming introduced by Linearbandkeramik, making it possible for previously established groups to thrive, and creating a persistent but permeable boundary that allowed transfer of ideas and low-level gene flow. This changed with the formation-through-mixture of Bell Beaker using populations [~]2500 BCE by fusion of local Rhine-Meuse people (9-17%) and Corded Ware associated migrants of both sexes. Their expansion from the Rhine-Meuse region then had a disruptive impact across a much wider part of northwest Europe, including Britain where its arrival was the main source of a 90-100% replacement of local Neolithic peoples.

genomics↗

bioRxiv2025

Mapping Dynamic Metabolic Energy Distribution in Brain Networks using fMRI: A Novel Dynamic Time Warping Framework

Schizophrenia has long been linked to impaired coordination of brain activity, yet most frameworks overlook two key dimensions: the amplitude of brain signals and the differing timescales on which regions operate. These factors are critical in disorders where neural activity is exaggerated and slowed. In healthy adults, networks compensate for mismatched processing speeds to maintain proportionate activity, but this process is poorly understood in schizophrenia. We developed a timescale-aligned, time-resolved framework that separates temporal distortions from genuine amplitude differences, enabling measurement of amplitude balance between networks across timescales. This approach was applied to large-scale fMRI datasets, including the Human Connectome Project and a multi-site schizophrenia cohort. Patients with schizophrenia showed greater amplitude imbalance, especially during fast fluctuations, along with more frequent re-entry into unbalanced states and slower recovery to stable coordination. We further identified a flexible intermediate state that patients occupied more often, and that predicted better working-memory performance. Across cohorts, amplitude imbalance was associated with greater symptom severity and poorer reasoning ability. These findings provide a new mechanistic view of dyscoordination in schizophrenia grounded in timescale-normalized amplitude dynamics, highlight aberrant recovery of amplitude balance as a core feature of the illness, and suggest that timescale-aligned amplitude imbalance may serve as a promising target for biomarker development.

neuroscience↗

bioRxiv2025

Epidermal Langerhans Cells Drive Painful Diabetic Neuropathy in a Sex-dependent Manner

The interaction between non-neuronal cells and nerve endings in the epidermis significantly influences the development of various diseases, including Painful Diabetic Neuropathy (PDN). PDN is a common and challenging complication of diabetes, characterized by changes in skin innervation accompanied by neuropathic pain. While there is growing evidence that epidermal non-neuronal cells, such as resident immune cells, play a crucial role in the progression of PDN, the underlying mechanisms of this neuropathy remain poorly understood. In our studies, we utilized transgenic methods, pain behavioral assessments, and single-cell RNA sequencing (scRNAseq) in a clinically relevant high-fat diet (HFD) mouse model of PDN and skin biopsy samples from PDN patients, to investigate the role of epidermal Langerhans cells (LCs) in this condition. We observed an increased density of LCs in the skin of PDN male mice coinciding with the onset of mechanical allodynia. Furthermore, we found that LCs density correlated with small fiber degeneration in HFD mice and skin biopsies taken from well-characterized PDN patients compared to healthy controls. Importantly, the selective ablation of LCs using a diphtheria-toxin strategy was found to prevent nociceptive behavior in male mice. This indicates that LCs are both necessary and sufficient for the development of mechanical allodynia and spontaneous pain in HFD male mice. Interestingly, when LCs were ablated in HFD female mice, it did not prevent but rather promoted pain behavior, suggesting the existence of sex-specific mechanisms mediated by LCs. scRNAseq transcriptomic analysis of the paw epidermis from HFD mice, which included both males and females, revealed significant sex-mediated differences in the expression of specific target genes within this PDN model. In male mice, scRNAseq identified differentially expressed genes associated with axonal guidance and immune responses in LCs. Additionally, by integrating single-cell RNA data from the epidermis and dorsal root ganglia (DRG) of male mice, we uncovered altered communication between LCs and cutaneous afferents through Semaphorin-Plexin signaling pathways in PDN. These findings highlight LCs as key contributors to the development of PDN and suggest their potential as therapeutic targets for innovative treatments, particularly topical therapies aimed at modulating immune cell activity and neuroimmune communication in the skin. Our investigations of male and female mice indicate that LCs may play different roles in mechanical sensation under both normal and pathological conditions, such as PDN. This underscores the importance of considering sex differences when developing more effective treatments. BRIEF SUMMARYThe interaction between non-neuronal cells and nerve endings in the epidermis plays a significant role in the development of Painful Diabetic Neuropathy (PDN), a challenging complication of diabetes. Using transgenic methods, pain behavioral assessments, and single-cell transcriptomics in a clinically relevant high-fat diet (HFD) mouse model of PDN, and in skin tissues from PDN patients, we identified epidermal Langerhans cells (LCs) as key contributors to the development of PDN. In these studies, we revealed, for the first time, a dimorphic role for LCs in the development of mechanical allodynia and spontaneous pain in a HFD model of PDN. Single-cell RNA sequencing (scRNAseq) of the paw epidermis from both male and female HFD mice identified sex-mediated differences in the expression of specific target genes within this PDN model. In male mice, scRNAseq of LCs revealed differentially expressed genes related to immune responses and axonal guidance, including Semaphorin-Plexin signaling pathways.

neuroscience↗