Biology subjects

Zhang, Z.

Publications and source records attributed to Zhang, Z..

At least 217 records · Page 12Linked to original sources

bioRxiv2026

Precision mapping and molecular contextualization of surgical outcome epicenters in temporal lobe epilepsy

AO_SCPLOWBSTRACTC_SCPLOWTemporal lobe epilepsy is the most common drug-resistant epilepsy, with surgical resection offering the primary path to seizure freedom. Despite standardized approaches, a substantial proportion of patients experience seizure recurrence, and the neurobiological substrates underlying these divergent outcomes remain unclear. We applied an individualized normative modeling framework to multimodal preoperative MRI data in a group that subsequently underwent surgical resection, to characterize patient-specific structural deviations and identify disease epicenters. Patients who became seizure-free exhibited spatially coherent abnormalities localized to the hippocampus and ipsilateral association regions, anchored in agranular limbic territories and enriched for genes linked to calcium-dependent signaling. Non-seizure-free patients, on the other hand, showed a more heterogeneous and distributed pattern of deviations, consistent with a "temporal-plus" network organization, and broader neuromodulatory dysregulation. Crucially, overlap between resected tissue and network-defined epicenters was closely associated with seizure freedom, independent of total resection volume. These findings provide a multiscale framework for precision surgical planning, shifting the focus from standardized tissue removal to targeted disconnection of patient-specific pathological hubs.

neuroscience↗

bioRxiv2026

Exploring genetic, expression and regulatory patterns of parental alleles in Muscovy duck (Cairina moschata) using haplotype-resolved assemblies

BackgroundAlthough the biological mechanism for heterosis has been debated for a long time, heterosis is widely utilized to increase the global productivity of crops and livestock. Recently, the mechanism has been well characterized in crops and livestock with a male-heterogametic XY system due to genomic assembly advancements, especially the availability of haploid genomes. However, the biological mechanism for heterosis remains unclear in poultry possessing the female-heterogametic ZW system. ResultsHere, we assembled chromosome-level diploid and haploid genomes of the Muscovy duck. We developed an efficient and cost-effective method to assemble 12 variation graph-haploid Muscovy duck genomes from three full-sibling pairs with high quality using short-read Illumina sequences. We further characterized genetic, expression and regulatory patterns of parental alleles at multiple scales. We found that maternal haploid genomes generally had more open chromatin organization and higher accessibility, and higher levels of gene expression, while showing similar DNA methylation levels when compared to paternal haploid genomes. In contrast, the female paternal Z chromosome showed the most, and the male paternal Z chromosome presented more, relaxed chromatin organization and chromatin accessibility, and gene expression compared to the male maternal Z chromosome. Thus, the ZW system largely relies on compensation and balance to regulate gene expression on the sex Z chromosome. Moreover, we identified non-Mendelian regions covering 0.26% of the genome ([~]3.18 Mb). These regions contained lower gene density, GC content, and repeat sequence frequency, but were enriched for DNA motifs bound by transcription factors, likely leading to a compacted chromatin structure and lower chromatin accessibility. ConclusionsOur work here provides a comprehensive profile of parental alleles genetic, expression and regulatory patterns in the female-heterogametic ZW system, and might be useful for the utilization of heterosis in poultry.

genomics↗

bioRxiv2026

ATP-independent unfolding of ubiquitin by Ufd1 initiates Cdc48/p97-mediated substrate processing

The Cdc48 ATPase (p97 or VCP in mammals) cooperates with its cofactors Ufd1 and Npl4 to extract polyubiquitinated proteins from membranes or multi-subunit complexes, promoting their proteasomal degradation. A ubiquitin molecule in the chain is unfolded in an ATP-independent manner and initiates substrate translocation through the central pore of the ATPase. How ubiquitin is unfolded remains unclear. Here, we demonstrate that the UT3 domain of Ufd1 interacts simultaneously with two Lys48-linked ubiquitins and unfolds one of the ubiquitins by binding its C-termianl {beta}-strand into a cleft. This unfolded ubiquitin is subsequently captured by Npl4 and Cdc48/p97. Experiments in vitro and in cells show that unfolding-defective mutants of Ufd1 indeed compromise Cdc48/p97 function. Our results explain how simple protein-protein interactions cause the unfolding of the remarkably stable ubiquitin. We also establish a linkage-specific recognition that involves ubiquitin unfolding.

biophysics↗

bioRxiv2026

Integrative Multi-omics Analysis of the Human Skeletal Muscle Response to Endurance or Resistance Exercise: Findings from the Molecular Transducers of Physical Activity Consortium (MoTrPAC)

The Molecular Transducers of Physical Activity Consortium (MoTrPAC) was established to systematically characterize the molecular basis of the health benefits of exercise. Here, we present the integrative, multi-omics response of human skeletal muscle to acute endurance (EE) and resistance (RE) exercise. Distinct temporal responses were observed, with changes in ATAC-seq, phosphoproteome, and metabolome occurring before changes in the transcriptome and proteome. These distinct temporal multi-omic dynamics were used to identify transcriptional regulatory hubs converging around MEF2A and NFIC regulation of autophagy, angiogenesis and metabolism. Further, early RE-specific phosphoproteome signatures counteracted epigenetic modifications and downregulated transcripts involved in protein turnover. Additional findings include suppression of HIPK2/3 kinase signatures linked to the acute exercise regulation of sarcomeric proteins TTN, NEB, ANKRD2 and LMOD2. Our data demonstrate distinct temporal regulation across the multi-omic landscape of human skeletal muscle, with EE and RE eliciting common and unique molecular signatures.

systems biology↗

bioRxiv2026

Constructing a single-objective oblique plane microscope (OPM) for fast, multi-colour, high-resolution volumetric fluorescence imaging

Oblique plane microscopy (OPM) is a light sheet microscopy technique that uses a single high numerical aperture (NA) objective for both illuminating the sample and collecting emission fluorescence from a tilted plane within the specimen. OPM has become indispensable in biological and biomedical research, providing rapid, high-resolution volumetric fluorescence imaging of live cells and tissues while minimising phototoxicity and photobleaching. It also overcomes the sample mounting challenges associated with conventional light sheet microscopes that require two orthogonally placed objectives. However, the application of OPM has been limited by the complex design and the intricate optical alignment and characterisation needed, particularly with the remote-refocusing system (RFS) in the emission path. This protocol offers a detailed, step-by-step guide for constructing an OPM setup using commercially available components and for characterising its performance to ensure optimal imaging quality. We aim to deliver the unique merits of OPM to researchers in life science and medicine, enabling them to visualise the spatiotemporal organisation of key biomolecules, structures, and cells in 3D at high resolutions.

biophysics↗

bioRxiv2026

gSV: a general structural variant detector using the third-generation sequencing data

Structural variants (SVs) are major contributors to genome diversity and disease susceptibility, particularly in cancer. Although third-generation sequencing technologies have substantially improved SV detection sensitivity, accurate detection of complex SVs remains challenging due to fragmented and heterogeneous alignment signals, as well as the dependence of many existing methods on predefined variant models. In this paper, we propose gSV, a general SV detector that integrates alignment-based and assembly-based approaches with the maximum exact match (MEM) strategy, with particular emphasis on resolving SVs with complex or atypical alignment signatures. Without predefined assumptions about SV types, gSV captures diverse variant signals, enabling the detection of SVs that are usually missed by conventional tools. Benchmarking using both simulated datasets and real long-read sequencing data demonstrates that gSV achieves improved sensitivity and overall detection performance compared with current state-of-the-art SV callers, particularly for simple and complex SV events with complex alignment patterns. Unique SV discoveries in four breast cancer cell lines, particularly in cancer-associated genes, demonstrate the potential biological relevance of gSV-enabled discoveries. Furthermore, analysis of a breast cancer cohort from the Chinese population highlights the utility of gSV for population-scale genomic studies. Collectively, gSV provides a unified framework for comprehensive SV discovery in both research and clinical genomics settings. O_TEXTBOXKey PointsO_LIExisting structural variant (SV) detection tools are limited in resolving SVs with complex alignment patterns due to their reliance on predefined variant models. C_LIO_LIgSV integrates alignment-based and assembly-based evidence using a maximum exact match (MEM) strategy, enabling capture of diverse and complex SV signals. C_LIO_LIBenchmarking on simulated and real long-read sequencing datasets demonstrates that gSV achieves competitive performance on canonical SV classes and improved sensitivity for complex SV patterns. C_LIO_LIApplication of gSV to breast cancer cell lines and a population-scale breast cancer cohort reveals previously unresolved SVs in cancer-associated genes, highlighting its utility in genomic and clinical studies. C_LI C_TEXTBOX

bioinformatics↗

bioRxiv2026

Exercise modulation of the alternative splicing landscape in human tissues

The diverse health benefits of exercise are associated with multi-organ molecular responses. Alternative RNA splicing (AS) is an important determinant of transcriptome and proteome diversity. We profiled the temporal effects of acute endurance and resistance exercise on the AS landscape of human skeletal muscle, adipose tissue, and blood, and studied regulatory mechanisms through integrated multi-omic analyses. We identified 5102 distinct differential AS (DAS) events, with the majority modifying protein-coding sequence (89%) and being independent of altered RNA expression (67%). Endurance and resistance exercise induced differing patterns of AS alterations with divergent temporal trajectories. We inferred the DAS-associated RNA-binding and DNA-binding proteins. In skeletal muscle, where DAS events were the most abundant, DAS genes were enriched for muscle structure- and RNA splicing-related processes, and splicing machinery components were regulated at the protein phosphorylation, RNA, and AS levels. These findings implicate AS regulation as a major mediator of the responses to exercise.

systems biology↗

bioRxiv2026

Myofibroblast lineage mapping and inhibiting subretinal fibrosis by targeting SMAD3 and MRTF pathways via microRNA-24 functional study

Subretinal fibrosis underlies the end-stage pathogenesis of retinal diseases including age-related macular degeneration (AMD). It can disrupt retinal structure and eventually lead to legal blindness by generating contractile force, fibrotic scarring, subretinal hemorrhage, and retinal detachment. Myofibroblasts are the predominant cells critically involved in subretinal fibrosis, however, the cellular contribution to myofibroblasts remains unclear. Here we demonstrate that multiple cell lineages, including macrophages, endothelial cells (EC), retinal pigment epithelial (RPE) cells and pericytes, significantly contribute to myofibroblasts in a laser-induced subretinal fibrosis model. We found microRNA miR-24 is significantly downregulated in the plasma of wet AMD patients. Overexpression of miR-24 represses epithelial-mesenchymal transition (EMT), endothelial-mesenchymal transition (EndMT), and the resulting fibrosis by regulating TGF-{beta}/SMAD3 and PAK4/LIMK2/MRTF pathways. Consistently, a combination of SMAD3 and MRTF inhibitors show superior efficacy to individual inhibitors in repressing fibrosis in vitro and laser-induced subretinal fibrosis in vivo. Together, these suggest the contribution of multiple cell-types in myofibroblast transformation in subretinal fibrosis, and repression of miR-24-regulated TGF-{beta}/SMAD3 and PAK4/LIMK2/MRTF pathways in multiple cell types holds therapeutic potential for treating subretinal fibrosis in AMD and other fibrotic disorders.

molecular biology↗

bioRxiv2026

Improved prediction of virus-human protein-protein interactions by incorporating network topology and viral molecular mimicry

The protein-protein interactions (PPIs) between viruses and human play crucial roles in viral infections. Although numerous computational approaches have been proposed for predicting virus-human PPIs, their performances remain suboptimal and may be overestimated due to the lack of benchmark dataset. To address these limitations, we first constructed a carefully curated benchmark dataset, ensuring non-overlapped PPIs and minimum sequences similarity of both human and viral proteins in the training and test sets. Based on this dataset, we developed vhPPIpred, a machine learning-based prediction method that not only incorporated sequence embedding and evolutionary information but also leveraged network topology and viral molecular mimicry of human PPIs. Comparative experiments demonstrated that vhPPIpred outperformed five state-of-the-art methods on both our benchmark dataset and three independent datasets. vhPPIpred also achieved high computational efficiency, requiring relatively low runtime and memory. Finally, vhPPIpred was demonstrated to have great potential in identifying human virus receptors, and in inferring virus phenotypes as the virus-human PPIs predicted by vhPPIpred can be used to effectively infer virus virulence. In summary, this study provides a valuable benchmark dataset and an effective tool for virus-human PPI prediction, with potential applications in antiviral drug discovery, host-pathogen interaction research and early warnings of emerging viruses.

bioinformatics↗

bioRxiv2026

miREA: a network-based tool for microRNA-oriented enrichment analysis

MicroRNAs (miRNAs) regulate gene expression at the post-transcriptional level, yet interpreting their function at the pathway level remains challenging. Existing enrichment analysis tools predominantly adopt network node-centric approaches that focus on gene expression profiles, neglecting the regulatory information encoded in miRNA-gene interactions (MGIs) that constitute network edges. This omission introduces analytical bias and limits biological interpretability, underscoring the need for network edge-based enrichment analysis methods that explicitly incorporate MGIs. Therefore, we present miREA, a network-based tool for miRNA enrichment analysis that leverages MGIs to characterize miRNA function at the pathway level. miREA contains five edge-based enrichment methods that integrate paired miRNA-gene expression and interactome profiles with pathway networks to perform MGI overrepresentation, MGI scoring-based, network topology-aware, and network propagation analyses. Benchmarking across multiple cancer types shows that the edge-based methods outperform node-based methods in improving sensitivity to identify relevant pathways and biological interpretability while maintaining controlled false positive rates. We further demonstrate the utility of miREA in elucidating miRNA-gene-pathway regulatory mechanisms in bladder cancer. miREA is a versatile enrichment analysis tool that provides pathway-level interpretation of human miRNA function and facilitates mechanistic hypothesis generation for experimental validation. HighlightsO_LImiREA uses information in miRNA-gene interactions for miRNA enrichment analysis. C_LIO_LImiREA contains five new edge-based algorithms that integrate expression and interactome profiles with pathway networks to characterize miRNA function. C_LIO_LIThe edge-based methods are more sensitive than node-based methods at identifying cancer-relevant pathways and more effective at identifying cancer genes and miRNAs. C_LIO_LIWe perform systematic analysis of the miREA methods to show their robustness on 16 cancer datasets. C_LIO_LIWe present a case study that demonstrates the potential of miREA to elucidate miRNA-gene-pathway regulatory mechanisms in bladder cancer. C_LI

bioinformatics↗

bioRxiv2026

Exploring the mechanism of Panax Notoginseng in the treatment of skin wound based on network pharmacology and experimental verification

BackgroundWhen the skin wound defect is too large, it is difficult for the body to heal itself, and medical treatment is often needed. How to shorten the healing cycle and reduce the incidence of infection is a difficult problem faced by clinicians. Panax notoginseng(PN), a traditional Chinese medicine, can promote the absorption of inflammatory exudates, granulation tissue formation and epidermal proliferation, effectively inhibit the inflammatory reaction of wounds and promote the healing of skin wounds, but its molecular mechanism has not been fully clarified so far. Based on network pharmacology and animal experiments, this study explored the target and molecular mechanism of PN in the treatment of skin wound. MethodsThe active components and potential targets of PN were obtained from the Traditional Chinese Medicine System Pharmacology Database and Analysis Platform (TCMSP) and UniProt database, and the skin wound-related targets were obtained from the GeneCards database. The intersecting targets were filtered using Venny 2.1.0. The intersecting targets were imported into the STRING database to construct a protein-protein interaction (PPI) network. Cluster analysis was performed using the MCODE and CytoHubba plugins in Cytoscape 3.8.2 to obtain core functional network modules and the top 10 key target genes. The intersecting targets were subjected to KEGG and GO enrichment analysis using the DAVID 6.8 database (https://david.ncifcrf.gov/). The component-target-pathway network of PN in the treatment of skin wounds was constructed using Cytoscape 3.8.2 software. In the experimental verification phase, 48 Sprague-Dawley (SD) rats were randomly divided into a control group and a PN group, with 24 rats in each group. A full-thickness skin excision was performed to establish a wound model. The PN group received intraperitoneal injections of the drug, while the control group received an equivalent amount of saline. Wound area measurements were taken on days 1, 4, and 7 after model establishment. Histopathological changes in the injured area and the expression and localization of TNF-, IL-6, and IL-10 were observed through hematoxylin and eosin (HE) staining and immunohistochemical staining. Relative expression levels of the three factors were detected using quantitative real-time polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA). ResultsThis study identified 8 major active components, 156 targets, and 115 signaling pathways involved in the treatment of skin wounds in rats using PN. The top 10 core target genes included TNF, IL-6, and IL-10, primarily enriched in signaling pathways such as NF-{kappa}B, MAPK, and JAK-STAT. Animal experiments revealed that at 4 and 7 days post-injury, the wound area in the PN group was significantly smaller than that in the control group (P<0.05). HE staining showed reduced infiltration of neutrophils and inflammatory cells in the injury area at 7 days in the PN group, accompanied by more pronounced fibroblast proliferation and collagen secretion. Molecular detection indicated that TNF-, IL-6, and IL-10 positive reactants were mainly distributed in the cytoplasm and matrix of epidermal cells, inflammatory cells, and fibroblasts in the skin. qRT-PCR and ELISA results showed that TNF- expression in the PN group was significantly lower than that in the control group at 4 and 7 days (P<0.01). IL-6 expression was lower than that in the control group at all time points, peaking at 4 days and then decreasing (P<0.01). IL-10 expression was significantly lower than that in the control group at 1 and 7 days (P<0.01). ConclusionPN exhibits characteristics such as multi-component, multi-target, multi-pathway synergy, and multiple regulatory pathways in the treatment of skin wounds. It can reshape the dynamic balance of cytokine networks, including TNF-, IL-6, and IL-10, optimize the temporal progression of "inflammation initiation - repair transition - tissue remodeling", and provide a therapeutic effect of "efficient debridement - orderly repair - low scar risk" for skin wounds. It is one of the ideal natural drugs for regulating skin wound healing.

bioinformatics↗

bioRxiv2026

Tmn blocks phage spread via plasmolysis and triggers synergistic defence responses

Membrane-associated phage defences remain poorly understood. Here we characterise Tmn, a YobI-family transmembrane P-loop NTPase that protects bacteria from phage infection by establishing a plasmolysis-associated antiviral state. Upon recognising phage T2 RIIB protein, Tmn enhances ATP turnover and selectively exports Mg2+, causing rapid cytoplasmic collapse that arrests phage replication without detectable membrane depolarisation or gross leakage of the cell content. Cryo-electron microscopy shows that Tmn assembles into a decameric membrane complex with extended cytosolic arms, an uncommon architecture among P-loop NTPases. The cytosolic arms, including a solenoid-like repeat domain, mediate trigger interaction and determine specificity. In addition to its primary defence function, Tmn-driven ATP collapse activates otherwise silent ATP-depletion-sensing defences, including Gabija and Septu type I, providing a mechanistic basis for synergy among defence systems and limiting secondary phage spread. These findings demonstrate that Tmn is a membrane-integrated sensor-effector that couples phage recognition to metabolic collapse and coordinated multi-layered immunity.

microbiology↗

bioRxiv2026

15-PGDH Inhibition Overcomes Muscle Regenerative Deficit Seen With GLP1-Receptor Agonist-Induced Weight Loss

Glucagon-like peptide-1 receptor agonists (GLP-1 RAs), including long-acting semaglutide, are revolutionary anti-obesity therapies. However, emerging evidence indicates that weight loss may come at the expense of skeletal muscle mass, a tissue essential for mobility, metabolic regulation, and overall health. Here, we show that an inhibitor of the gerozyme 15-hydroxyprostaglandin dehydrogenase (PGDHi), which boosts PGE2 levels, increases skeletal muscle mass, strength, and regeneration in the presence of semaglutide. We find that in a high fat diet-induced mouse model of obesity, semaglutide alone induces significant loss of muscle mass, while retaining contractile function. However, muscle regeneration and recovery of strength post-injury are hindered by semaglutide. This regenerative deficit is due to impeded stem cell function, which is overcome if mice are treated with a combination of PGDHi and semaglutide. Our data show that GLP-1-mediated weight loss interferes with this key muscle-building function, which PGDHi co-treatment counteracts to promote proper muscle regeneration and restored strength.

cell biology↗

bioRxiv2026

Endocardial TIE1 synergizes with TIE2 to regulate the atrial internal muscular network assembly

Atrial cardiomyopathy is characterized by altered atrial structures, yet its underlying genetic basis remains inadequately explored. TIE1 mutations are reported in lymphedema patients, but whether they exhibit cardiac defects alongside lymphatic abnormalities remains unknown. Here, single-cell RNA sequencing showed high Tie1 and Tek expression in atrial endocardial cells. TIE1 deficiency severely disrupted atrial morphogenesis, with only minor ventricular defects. Bulk RNA-seq showed reduced transcripts for endothelial development and cardiac trabeculation in Tie1 mutants. Separate RNA-seq analysis of the atria and ventricles further confirmed these findings, revealing a more pronounced downregulation of trabecular genes, including Tek, in the atria upon Tie1 loss. Endothelial deletion of Tek impaired trabeculation, particularly in atria. Consistently, Tie1 loss combined with a single null Tek allele disrupted both atrial and ventricular trabeculation. Strikingly, atrial morphogenesis defects appeared within 48 hours of induced endothelial Tie1 loss coupled with Tek heterozygous deletion, implying a critical endocardial role in organizing the atrial muscular network. The synergy of TIE1 and TIE2 in remodeling atrial trabeculae was confirmed postnatally, whereas TIE1 insufficiency alone had no effect. Together, TIE1 is differentially required for atrial versus ventricular development, acting synergistically with TIE2 to regulate endocardial cell-coordinated assembly of atrial internal muscular network.

developmental biology↗

bioRxiv2026

Protein Phosphatase 2A Orchestrates Mitochondrial Dynamics via MAPK Signaling in heart

Heart as one high ATP consuming organ accounts for 5% of the total oxygen demands. The central question of heart health is how mitochondria fit its needs. Impaired mitochondrial dynamics (fission and fusion) have been observed in failing heart, but whether and how phosphorylation events involved in mitochondrial quality control are still imperceptive. The phosphatase 2A catalytic subunit (PP2A c) cardiac-specific knockout mouse (KO), which exhibited a hypertrophic cardiomyopathy phenotype, was studied. We profiled the pattern of morphological and functional alteration of cardiac mitochondria that appeared during postnatal development. Increased heterogeneity of mitochondria and a decreased ATP yield was displayed. Notably, a fission procedure escalated. To illustrate the protagonist of the mitochondrial dynamics, we applied a high-throughput spectrometry-based phosphoproteomic screening following by GO and KEGG pathway annotations for 788 phosphosites, accounting for 90 proteins. Results suggested that the MAPK signaling may be a predominant factor associated with those mitochondrial alternations in KO hearts. Furthermore, we identified hyperphosphorylated ERK2 accumulated into the nucleus regarding PP2Ac depletion. Consequently, Fis1 expression was accelerated at the transcriptional level which facilitated recruitment of Drp1 onto the outer mitochondrial membrane. The mitochondrial fission towards shifting led to excessed mitophagy and is considered the culprit in early mortality. These findings are indicative of the fundamental role of PP2A in mitochondrial dynamics regulation and cardiomyopathy progression. During the progression of heart failure, the phospho-regulation of ERK2 could be a novel therapeutic approach to prevent or attenuate adverse hypertrophic cardiomyopathy.

cell biology↗

bioRxiv2026

Thermal Nano-Engineering of Ginger Extracellular Vesicles for Targeted Oral Therapy of Colitis

Plant-derived extracellular vesicles (PEVs) are promising candidates for oral drug delivery, yet their clinical translation is hindered by limited targeting precision and inconsistent systemic absorption. While surface engineering can enhance tissue accumulation, strategies that preserve biocompatibility and enable scalable production remain limited. Here, we introduce a simple thermal processing approach, boiling, to structurally reconfigure ginger EVs into functionally enhanced, thermally reassembled nanoparticles (B-GEVs). The surface architecture of B-GEVs is enriched with key vesicle trafficking regulators, including V-type proton ATPase subunit G, ARF1, and {beta}-adaptin-like protein. This specific composition drives their tissue-specific accumulation in the intestine and liver and potentiates clathrin-dependent cellular uptake in intestinal cells by 8.57-fold. Beyond superior intrinsic anti-inflammatory activity through NLRP3 inflammasome suppression, B-GEVs function as an efficient oral delivery platform. When loaded with TNF- siRNA, they enable a synergistic therapy that simultaneously modulates upstream inflammation and silences key downstream mediators, showing potent efficacy in colitis. Our findings position boiling as a natural strategy for enhancing the bioactivity and targeted oral delivery potential of ginger-derived EVs.

bioengineering↗

bioRxiv2026

Neural Arming Niche in Tumor-Draining Lymph Nodes Programs CD8⁺T Cell Cytotoxicity via GZMB Norepinephrinylation

Lymph nodes are densely innervated neuro-immune hubs where naive T cells are primed and "armed" with cytotoxic machinery, yet how local neural cues in tumor-draining lymph nodes (tdLNs) set CD8 T cell effector reserves remains unclear. Here we show that tdLN-local sympathetic norepinephrine (NE) instructs CD8 T cell cytotoxic capacity by stabilizing the granzyme B (GZMB) pool through covalent norepinephrinylation. Exercise, used as a physiological perturbation, increases GZMB selectively in tdLN and tumor CD8 T cells without altering T cell abundance or cytokine output; tdLN sympathectomy abolishes these effects and reduces tdLN NE enrichment. Mechanistically, NE enters CD8 T cells and norepinephrinylates GZMB at Gln43, limiting UHRF1-mediated ubiquitination and proteasomal degradation. Genetic attenuation of norepinephrinylation destabilizes GZMB, impairs cytotoxicity, and accelerates tumor growth. Brief ex vivo NE conditioning increases GZMB reserves and improves adoptive tumor control.

cancer biology↗

bioRxiv2026

Leaf age modulates physiological and metabolic responses to contrasting nitrogen forms in Chinese fir (Cunninghamia lanceolata (Lamb.) Hook)

Atmospheric nitrogen (N) deposition is increasingly affecting global ecosystems, with nitrate contributing a growing proportion alongside ammonium. However, the interaction between N forms and leaf developmental stage in shaping physiological and metabolic strategies in Chinese fir remains poorly understood. In this study, a field experiment was conducted to explore the physiological and metabolic responses of young and old leaves to ammonium and nitrate N addition. Our findings showed that N addition enhanced photosynthetic performance in young leaves, with a stronger effect from nitrate. In contrast, old leaves exhibited limited photosynthetic response but accumulated higher non-structural carbohydrates and showed elevated N assimilation enzyme activities, particularly under nitrate addition. Phytohormone profiles varied between leaf ages, with young leaves having higher auxin levels while old leaves exhibiting increased abscisic and salicylic acid contents under N addition. Additionally, N addition induced differential reprogramming of amino acid metabolism, with age-dependent accumulation patterns. Metabolomic analysis identified key amino acids involved in coordinating carbon-nitrogen metabolism. These results highlighted the complementary metabolic strategies by young and old leaves of Chinese fir under contrasting N forms addition and emphasized the importance of considering both N form and leaf age in optimizing N management for sustainable plantation practices. HighlightsO_LINitrate enhanced photosynthesis in young Chinese fir leaves more effectively than ammonium. C_LIO_LIOld leaves prioritized C storage and N assimilation under N addition, especially nitrate. C_LIO_LIComplementary metabolic strategies between leaf ages optimized resource use under different N forms addition. C_LI

plant biology↗