Biology subjects

Zhang, J.

Publications and source records attributed to Zhang, J..

At least 235 records · Page 13Linked to original sources

bioRxiv2026

Serine Stabilizes SLC7A11 and Enhances Cystine Influx to Protect Against Acute Pancreatitis

Lethal sterile inflammatory diseases are linked to amino acid metabolism, but the role of serine remains unclear. Here, we show that dysregulated serine metabolism and reduced plasma serine levels correlate with disease severity of acute pancreatitis (AP) in patients and mouse models. Elevating serine levels via exogenous serine supplementation or pancreatic phosphoglycerate dehydrogenase (PHGDH) overexpression mitigates pancreatic injury, whereas a serine deprivation diet or pancreatic PHGDH knockdown exacerbates AP. Serine prevents cell death and oxidative stress in pancreatic acinar cells, human induced pluripotent stem cells-derived pancreatic organoids and mouse pancreatic tissue. Serine enhances cysteine and glutathione biosynthesis primarily by promoting solute carrier family 7 member 11 (SLC7A11)-dependent cystine uptake rather than by serving as a direct substrate. Mechanistically, the E3 ubiquitin ligase NEDD4 mediates ubiquitination and degradation of SLC7A11, whereas serine binds to NEDD4 and thereby inhibits SLC7A11 degradation. Similarly to serine, pharmacological inhibition of NEDD4 alleviates lipid peroxidation and pancreatic injury. These findings identify serine as a critical signaling regulator of SLC7A11 stability and oxidative stress, and provides a new therapeutic strategy for AP and associated sterile inflammatory disorders. HighlightsAcute pancreatitis (AP) is linked to abnormal serine metabolism and serine depletion. Serine prevents cell death in AP acinar cells, human pancreatic organoids and mice. Serine promotes SLC7A11-dependent cystine uptake and glutathione levels in acinar cells. Serine reduces NEDD4-mediated ubiquitination of SLC7A11. In briefSerine protects against cell death and pancreatic injury in acute pancreatitis by stabilizing SLC7A11 through disruption of NEDD4-mediated ubiquitination in acinar cells.

pharmacology and toxicology↗

bioRxiv2026

Reactivation of a TAL1 progenitor cell enhancer region by non-coding somatic variants in T-lineage acute lymphoblastic leukemia

Aberrant activation of TAL1, a key oncogenic driver, defines a major subgroup comprising [~]30% of childhood T-lineage acute lymphoblastic leukemias (T-ALLs). We and others have shown that somatic non-coding mutations within upstream and intronic cis-regulatory regions of TAL1 contribute to transformation by creating binding sites for MYB and other transcription factors. Here we investigated cis-regulatory mechanisms mediated by somatic mutations occurring in an intergenic region located 29 kilobase pairs downstream of the canonical TAL1 transcription initiation site, implicated in 6% of TAL1-expressing T-ALLs. These somatic variants include i) complex indels resulting in de novo MYB transcription factor binding sites (TFBSs) and ii) internal tandem duplications (ITDs) encompassing canonical MYB TFBSs. Chromatin immunoprecipitation sequencing (ChIP-seq) revealed binding of the TAL1 core regulatory circuit (CRC) transcription factors MYB, GATA3, and RUNX1, resulting in enhancer activity mediated by sequences with the mutant allele. Strikingly, ChIP-seq peaks for the repressive H3K27me3 mark and the active H3K27ac mark co-existed across TAL1 regulatory sequences but enriched for different haplotypes. TAL1 transcription from the mutant haplotype initiated from a promoter located within exon 4 of the canonical TAL1 transcript, resulting in a short isoform normally expressed by hematopoietic stem cells (HSC). Interestingly, neither the isoform expression nor the enhancer activity could be predicted by the sequence-to-function deep learning artificial intelligence (AI) model AlphaGenome, emphasizing the importance of experimental validation. Our findings indicate that selection for cis-regulatory, non-coding variants leads to reactivation of enhancers normally active in HSC but silenced in differentiated lineages during normal hematopoietic cell development.

cancer biology↗

bioRxiv2026

Comparative single-cell transcriptomic roadmap of mammalian fetal ovarian development

Early ovarian development establishes the cellular basis of female reproduction, yet its cellular composition and developmental dynamics remain poorly characterized across mammalian species. Here, we generated a cross-species single-cell transcriptomic atlas of early female gonadal development in cattle (E38-E112), human (PCW6-16), and mouse (E11.5-E18.5), integrating 107,930 cells across comparable developmental windows, spanning sex determination and early ovarian differentiation. We identified 11 shared gonadal cell types across three species, including germ cells and granulosa cells, and uncovered a previously undescribed bovine-specific cell population with steroidogenic features, suggesting species differences in ovarian somatic lineage development. Epigenetic regulators exhibited dynamic activation in germ cells and granulosa cells. Developmental trajectory analysis revealed shared and species-specific genes associated with germ cell and granulosa cell development, including conserved dynamically expressed genes such as TFAP2C and ZCWPW1 in germ cells and FOS and JUNB in granulosa cells. Cross-species cell type classification using a machine learning support vector machine (SVM) model revealed transcriptionally conserved cell types, such as immune cells and germ cells, while granulosa cells showed substantial cross-species divergence, associated with extracellular matrix-related gene expression. Together, our study provides a comparative framework for understanding conserved and divergent mechanisms of early ovarian development across mammals.

developmental biology↗

bioRxiv2026

IL-36γ Drives Th9 cell Differentiation via IκBζ to Sustain Antitumor Immunity

Th9/Tc9 cells polarized with TGF-{beta} and IL-4 exhibit superior antitumor efficacy, prompting extensive efforts to optimize their differentiation protocols and augment therapeutic potency. In the current study, we identified IL-36{gamma} as a potent cytokine that synergizes with TGF-{beta} to robustly drive Th9 differentiation both in the presence and absence of IL-4. IL-36{gamma}-programmed Th9 cell subsets exhibited phenotypic and transcriptional profiles identical to classic Th9 cells. Mechanistically, IL-36{gamma} drove Th9 cell differentiation through amplifying key signaling pathways such as STAT6, STAT5, and NF-{kappa}B that are essential for classic Th9 programming. Notably, we uncovered a novel regulatory axis wherein IL-36{gamma} upregulates the transactivation factor I{kappa}B{zeta}, which directly governs Th9 lineage specification. In in adoptive cell therapy (ACT) models, IL-36{gamma}-polarized Th9 cell subsets demonstrated enhanced antitumor efficacy, attributable to their sustained persistence, reduced exhaustion markers and stem-like/memory properties. Collectively, this study elucidates a previously unrecognized I{kappa}B{zeta}-dependent mechanism underpinning Th9 differentiation and highlights the translational potential of IL-36{gamma}-engineered Th9 cells as a valuable ACT strategy for refractory tumors.

immunology↗

bioRxiv2026

Guidance for high-quality functional gene embeddings from large language models

Large language models (LLMs) are increasingly used to generate gene embeddings, yet systematic benchmarks of prompting strategies and practical guidance for obtaining biologically meaningful representations remain limited. Here we present GEbench, an evaluation framework for assessing LLM-derived gene embeddings across different tasks, prompting strategies, and LLM architectures. GEbench revealed that embedding quality depends primarily on whether the input text contains explicit functional information, rather than on sparse gene identifiers or model size. Identifier-based embeddings showed weak biological organization, whereas embeddings derived from functional descriptions consistently achieved stronger functional separation and predictive performance. Notably, Self-Des, which extracts embeddings from model-generated gene function descriptions, enabled locally deployable LLMs to generate high-fidelity representations that approach the quality of expert-curated databases. Genome-scale analyses further supported these findings, indicating that explicit functional descriptions are an effective design principle for generating high-quality gene embeddings from LLMs.

bioinformatics↗

bioRxiv2026

Natural Variations of Spring Wheat Nitrogen and Carbon Assimilation under Different Inorganic Nitrogen Forms and CO2 levels

Wheat (Triticum aestivum L.) production depends upon nitrogen fertilization and may be threatened by the climate conditions anticipated in the next few decades. The natural variation in the ability of wheat to assimilate carbon and different inorganic nitrogen forms, nitrate (NO3-) and ammonium (NH4+), into vegetative growth remains unexplored. Here, we evaluated growth under either NO3- or NH4+ as a sole nitrogen source for 875 spring hexaploid wheat accessions that represent the genetic diversity within the global germplasm. These accessions varied over 8-fold in vegetative biomass but grew similarly under moderate levels of either nitrogen form. At high, potentially toxic concentrations of NH4+, however, they lost approximately 20% of their biomass. We characterized the influence of changing CO2 levels in bi-parental Nested Association Mapping populations. Genetic backgrounds determined wheat biomass responses to CO2 enrichment and nitrogen form. Genome-Wide Association and linkage mapping identified certain loci as consistently associated with biomass accumulation under different nitrogen forms and CO2 levels. These results will assist breeding efforts to develop food crops that are resilient to the worlds climate changes.

plant biology↗

bioRxiv2026

Inhibition of oogenic JNK preserves fertility and ovarian hormones during DNA-damaging cancer therapy

Primary ovarian insufficiency (POI) and related infertility, early menopause, and endocrine disorders due to hormonal deficiency are major side effects in young female cancer patients undergoing cancer therapy. Current strategies preserving the fertility and hormonal functions of the ovary remain imperfect due to concerns of feasibility, efficacy, or safety. Herein, we identified c-Jun N-terminal kinase (JNK) as a pivotal regulator of the DNA damage response (DDR) signaling in oocytes of primordial follicles in response to DNA-damaging cancer therapy. Using pharmacological JNK inhibition and a genetically modified mouse model with oocyte-specific JNK deletion, together with histological, bioinformatic, and molecular approaches, we demonstrated that JNK inhibition prevented chemotherapy-induced oocyte apoptosis and POI, and preserved long-term reproductive cycles and fertility. Mechanistically, JNK was activated in response to chemotherapy-induced DNA damage in oocytes of primordial follicles, causing activation of transcription factor TAp63 and subsequent oocyte apoptosis, ultimately resulting in diminished ovarian reserve and POI. A more clinically relevant breast cancer-bearing mouse model revealed that JNK inhibition preserved the ovarian reserve without compromising anti-cancer efficacy of chemotherapy. Together, our study identifies oocyte-intrinsic JNK as a promising target for developing ovarian protectants and safeguarding reproductive health and fertility in young female cancer survivors.

pharmacology and toxicology↗

bioRxiv2026

Spatial Multi-Omics Reveals Extracellular Matrix Remodeling and VSMC Phenotypic Switching in Moyamoya Disease

BackgroundMoyamoya disease (MMD) is a progressive cerebrovascular disorder characterized by steno-occlusive lesions and intimal hyperplasia. Although vascular smooth muscle cell (VSMC) phenotypic switching is implicated in its pathogenesis, the precise spatial interplay between extracellular matrix (ECM) remodeling and local metabolic alterations within the distinct vascular microenvironments remains unknown. MethodsSuperficial temporal artery (STA) samples from patients with MMD and controls were analyzed by histology, immunofluorescence, spatial transcriptomics, spatial proteomics, and spatial metabolomics. Single cell RNA sequencing was used to profile the cellular landscape of STA tissues. To functionally validate the identified pathway, human brain vascular smooth muscle cells (HBVSMCs) were stimulated with fibronectin 1 (FN1), and patient derived induced pluripotent stem cell smooth muscle cells (iPSC-SMCs) were generated for migration and protein expression assays following ITGA5 silencing or focal adhesion kinase (FAK) inhibition. ResultsMMD STA samples exhibited marked intimal hyperplasia with medial thinning and intimal accumulation of -SMA positive cells. Spatial transcriptomic and proteomic analyses identified an intimal remodeling program characterized by increased FN1, EFEMP1, fibronectin, ITGA5, and FAK, together with reduced MYH11. FN1 stimulation promoted smooth muscle cell migration, ECM associated protein expression, and FAK phosphorylation, whereas ITGA5 knockdown or FAK inhibition attenuated these effects. Patient derived MMD iPSC-SMCs showed similar abnormalities, including enhanced migration, increased FAK activation, reduced contractile markers, and increased ECM associated proteins. Spatial metabolomics and integrated multi-omics analyses further revealed that these changes were coupled to a metabolically depleted intimal niche enriched for reduced acyl-CoA related metabolites. ConclusionsIntegrated spatial multi-omics identifies coupled ECM remodeling and metabolic alteration in the hyperplastic intima of MMD. Within this context, the FN1-ITGA5-FAK axis emerges as a plausible mediator of smooth muscle remodeling that warrants further validation. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=165 SRC="FIGDIR/small/721225v1_ufig1.gif" ALT="Figure 1"> View larger version (90K): org.highwire.dtl.DTLVardef@3f6e19org.highwire.dtl.DTLVardef@554d49org.highwire.dtl.DTLVardef@451dd9org.highwire.dtl.DTLVardef@1aac1e9_HPS_FORMAT_FIGEXP M_FIG C_FIG

neuroscience↗

bioRxiv2026

A bifidobacterial enzyme orchestrates ecology and function of infant gut bacterial community

Human milk oligosaccharides (HMOs) are abundant and structurally diverse glycans that shape the development of infant gut microbiota. Yet, how individual HMOs and bacterial genes drive the community assembly remain elusive. Here, we reconstructed an eight-member infant Bacterial Community (iBaCo) from representing dominant taxa in human infant feces. When individual HMOs were the sole carbohydrate source, they showed deterministic effects on the iBaCo composition and metabolic output. Notably, the tetramer HMO lacto-N-tetraose (LNT), in spite of its identical monomer composition as lacto-N-neotetraose (LNnT), showed a strong effect on maintaining Bifidobacterium breve abundance in iBaCo, whereas LNnT did not. Monoculture growth profiling, proteomics, enzymatic kinetic assay, and molecular docking revealed that {beta}-galactosidase D4BMY8 and the relevant downstream pathways are induced by LNT and that D4BMY8 has substrate preference on LNT over LNnT, enabling a faster growth of Bi. breve and accumulation of acetate and lactate in LNT compared to LNnT. Metabolic flux analysis indicated that the substrate-preference of {beta}-galactosidase D4BMY8 drives the skewed energy cost toward lactate/acetate metabolic output. Finally, the D4BMY8-encoding gene lacZ5 is widely spread in all isolated Bi. breve genomes, but divergently distributed in infant metagenome-assembled Bi. breve genomes. Together, we demonstrated that a single enzyme-substrate interaction could orchestrate the composition and metabolic function of an infant bacterial community, which may contribute to the assembly of dynamic infant gut microbiota. Our integrative approach provides a mechanistic framework for understanding the interaction between diet, microbial community, and infant gut health.

microbiology↗

bioRxiv2026

TMEM106B C-terminal fragments drive nucleocytoplasmic transport failure and TDP-43 mislocalization in the aging human brain

TMEM106B is a lysosomal membrane protein and major genetic modifier of multiple neurodegenerative diseases, including frontotemporal lobar degeneration, Alzheimers disease, and amyotrophic lateral sclerosis. Proteolytically generated C-terminal fragments of TMEM106B assemble into amyloid fibrils that accumulate in the brains of individuals with neurodegenerative disease and in cognitively normal aged adults, yet how these fibrils produce neuronal dysfunction has remained unclear. Here, we show that cytosolic and lysosome-directed TMEM106B C-terminal fragments (CTF and gCTF) form detergent-insoluble amyloid aggregates, drive redistribution of endogenous TDP-43 from the nucleus to the cytoplasm, and accelerate neuronal death. Unbiased proximity proteomics identified the inner nuclear membrane LAP1-TorsinA axis as a fragment-specific interactome, and co-immunoprecipitation confirmed a direct physical interaction between gCTF and LAP1 that was not observed with full-length TMEM106B. Fragment expression disrupted Lamin B1 organization, mislocalized the nuclear import machinery KPNB1 and RanGAP1, and impaired importin-dependent nuclear transport in primary cortical neurons. Critically, neurons harboring endogenous TMEM106B fibrillar pathology in aged human frontal cortex exhibited the same phenotypes, namely disrupted Lamin B1 and LAP1 localization and cytoplasmic redistribution of TDP-43, whereas fibril-negative neurons from the same cases and younger control tissue retained intact nuclear envelope organization. These findings define TMEM106B proteinopathy as an upstream driver of nuclear envelope disruption and nucleocytoplasmic transport failure, linking a widespread feature of brain aging to a central mechanism of neurodegeneration.

neuroscience↗

bioRxiv2026

Genetic compensation in β-actin mutants occurs independently of mutations that destabilize β-actin mRNA

Proper maintenance of gene expression in response to mutations or environmental fluctuations is critical for cell development and survival. Recently, a novel genetic compensation mechanism was described wherein mutant mRNA decay triggers increased transcription of paralogous genes. This effect was reported for several genes, including {beta}-actin (Actb) in mouse embryonic stem cells, where Actb mRNA with a premature termination codon enhances transcription of its paralog, {gamma}-actin (Actg1), and partially rescues cytoskeletal defects. Here we show that, in both mouse and human embryonic stem cells, mutations in the ACTB gene, regardless of mutant mRNA expression, trigger genetic compensation. Furthermore, transgenic expression of mutant ACTB mRNA with a premature stop codon fails to induce genetic compensation. Depletion of the SRF or MRTF-A transcription factors, which are known to increase ACTB transcription in response to low ACTB protein levels, diminishes the genetic compensation response in ACTB mutants. These results suggest that genetic compensation in ACTB mutants is primarily mediated by a transcriptional feedback loop via SRF/MRTF-A, independent of the expression or degradation of mutant ACTB mRNA.

molecular biology↗

bioRxiv2026

TAR syndrome causal gene RBM8A is critical for embryonic bone development and proper Hedgehog signaling

RNA-Binding Motif Protein 8A (RBM8A) is a core component of the exon junction complex (EJC), which plays a fundamental role in post-transcriptional RNA regulation. Mutations in the RBM8A gene cause thrombocytopenia-absent radius (TAR) syndrome, characterized by reduced platelet number and radial defects. However, how its function contributes to bone development remains poorly understood. We found that RBM8A is highly expressed in human bone marrow-derived mesenchymal stem cells (MSCs) and mouse embryonic limb buds. To further investigate the role of RBM8A in bone development, we generated an Rbm8a conditional knockout mouse (cKO) model in the early limb bud and craniofacial mesenchyme using the Prx1-cre line, which recapitulates key features of TAR syndrome. Rbm8a deficiency in this mouse model resulted in significant defects in limb and craniofacial development, including shorter limb bones across developmental timepoints, delayed ossification, altered cranial morphogenesis, along with impairments in motor function and reduced brain size. To elucidate the molecular basis of these phenotypes, we performed RNA immunoprecipitation sequencing (RIP-seq) in embryonic limb tissue. Our analysis revealed that RBM8A preferentially associates with long, exon-rich transcripts with high isoform complexity. These transcripts are enriched in pathways related to primary cilia, RNA processing, and developmental signaling. Consistent with this, transcriptomic and enrichment analyses identified strong associations with ciliopathy-related and congenital skeletal disease gene sets. At the cellular level, Rbm8a deficiency resulted in altered expression of key chondrogenic and ossification markers in embryonic limb bone tissue, such as reduced SOX9 and increased IHH in the primary ossification center, suggesting impaired progression of endochondral ossification and disrupted Hedgehog pathway activity. Quantitative Reverse Transcription-PCR (qRT-PCR) analysis further validated selective downregulation of Hedgehog pathway targets, including Gli3 and Hhip, while other developmental genes remained unchanged, indicating pathway-specific attenuation. Together, these findings support a model in which Rbm8a regulates structurally complex transcripts that are enriched in ciliary and developmental signaling pathways, and that disruption of this process leads to selective impairment of Hedgehog signaling and skeletal development. These results provide a novel mechanistic link between RNA processing and developmental signaling and offer new insight into how defects in RNA regulation contribute to congenital skeletal abnormalities.

developmental biology↗

bioRxiv2026

OneGenomeRice (OGR): A Genomic Foundation Model for Rice

The transition of genomics to a predictive intelligence discipline is driven by the advent of genomic foundation models. While substantial progress has been observed in all-life and human-centric models, plant species, particularly for the staple crops, remains hindered by a lack of models. Here we introduce OneGenome-Rice (OGR), a rice (Oryza sativa) genomic foundation model pre-trained on a genomic dataset comprising 422 high-quality genomes of cultivated and wild rice. OGR is engineered upon a Mixture of Experts (MoE) transformer architecture with 1.25-billion parameters and supports an ultra-long context window of up to 1 million base (Mb) pairs at single-nucleotide resolution. A comprehensive benchmark demonstrated that OGR significantly outperforms existing state-of-the-art multi-plant or all-life genome models in 11 categories (e.g. motif identification, sweep detection, etc). We further demonstrated the utility of OGR in several downstream applications, such as indica-japonica subspecies introgression analysis, identification of agronomy trait-associated functional loci and prediction of gene expression from DNA sequences. These results establish OGR as a promising foundational computational infrastructure for rice functional genomics and precision breeding. The OGR and its fine-tuned models, including pretrained weights, training code and the rice genomic benchmark suit, have been fully opened.

genomics↗

bioRxiv2026

Revisiting the genome assembly of Lupinus species reveals differential diploidization after a shared whole-genome duplication

Accurate genome assemblies are essential for comparative genomics, yet Hi-C-guided scaffolding can introduce structural errors that misrepresent chromosome architecture and bias evolutionary inferences. Here, we identified pervasive scaffolding errors - including artificial fusions, internal inversions, and incomplete contig mounting - in two previously published Lupinus genomes (L. cosentinii and L. digitatus) using a segmentation method based on LTR retrotransposon density. We re-assembled both genomes, producing chromosome-level references of 472.7 Mb (16 chromosomes) and 427.2 Mb (21 chromosomes), with BUSCO completeness >98.5%. Synteny validation and re-application of LTR profiling confirmed that all prior errors were resolved. Using these corrected genomes together with four additional Lupinus species and two outgroup legumes, we investigated post-polyploid evolution. Synonymous substitution rate (Ks) analysis revealed a genus-specific whole-genome duplication (WGD) event (Ks = 0.17) shared by all six Lupinus species. The proportion of WGD-derived genes varied markedly, from 60% in L. digitatus to only 36% in L. mutabilis, indicating differential diploidization. While all species retained a core set of WGD duplicates enriched in cytoskeleton organization, ion transport, and defense responses, each exhibited lineage-specific functional trajectories: cell wall modification in L. cosentinii and L. digitatus, nitrogen metabolism in L. albus and L. angustifolius, flower development in L. luteus, and stress/lipid metabolism in L. mutabilis. Our corrected assemblies provide optimal references for Lupinus comparative genomics, and our findings demonstrate that a shared WGD event can lead to both conserved and highly divergent post-polyploid fates, likely underpinning adaptive diversification within the genus.

genomics↗

bioRxiv2026

Neuronal precursor cell persistence in Ganglioglioma is associated with ECM remodeling and immune cell infiltration

BackgroundGangliogliomas (GGs) are low-grade glioneuronal tumors that frequently present with drug-resistant epilepsy. Although their indolent course contrasts with their high epileptogenic potential, the oncogenic mechanisms sustaining neuronal precursor-like populations within the tumor microenvironment remain poorly defined. MethodsWe performed spatial transcriptomic profiling on eight histologically confirmed GGs and matched healthy cortex to map the cellular and molecular architecture of the tumor microenvironment. Integrated analysis with weighted gene correlation network analysis (WGCNA) defined recurrent oncogenic programs and spatially resolved tumor-stroma interactions. ResultsEight conserved gene modules emerged, encompassing physiological cortical, reactive glial, and oncopathological programs. The latter captured extracellular matrix (ECM) remodeling, vascular-immune signaling, and persistence of immature, proliferative neuronal-like states. Spatial modeling revealed that these oncopathological programs form structured niches at the tumor-brain interface, where radial glia-derived neuronal-like tumor cells coexist with immune and stromal elements engaged in ECM turnover and cytokine signaling. ConclusionsGanglioglioma represents a hybrid glioneuronal neoplasm in which developmental neuronal programs are co-opted by tumor-associated stromal and immune cues. This convergence establishes a permissive oncogenic niche that sustains precursor-like tumor cells and provides a mechanistic basis for both the tumors benign growth and its intrinsic epileptogenicity. Key PointsO_LISpatial transcriptomics identifies reproducible transcriptional programs that define the ganglioglioma microenvironment. C_LIO_LITumor-associated regions show transcriptional programs consistent with immature neuronal states together with ECM remodelling and immune activity. C_LIO_LISingle-cell reference data indicate that immature neuronal programs in ganglioglioma resemble radial glia-derived developmental states. C_LI Importance of the StudyGanglioglioma is a low-grade glioneuronal tumor that combines benign growth with pronounced epileptogenicity, yet the molecular basis of this dual behavior remains poorly understood. Through spatial transcriptomics integrated with single-cell analysis, we reveal that ganglioglioma architecture is defined by two interacting transcriptional axes: a residual glioneuronal network and a tumoral niche enriched for extracellular-matrix, vascular, and immune programs. Within these niches, immature neuronal-like tumor cells persist in a developmentally arrested state maintained by ECM-immune signaling. This spatially organized interplay between physiological and pathological programs explains both the low oncologic aggressiveness and high excitability of these lesions. Our findings provide molecular signatures that may refine diagnostic classification within the LEAT spectrum, delineate epileptogenic zones, and identify candidate pathways for therapeutic modulation of the ganglioglioma microenvironment.

neuroscience↗

bioRxiv2026

Serine proteases are required to activate influenza D virus haemagglutinin-esterase fusion (HEF) protein

Influenza D virus (IDV), the most recently identified member of the Orthomyxoviridae, was first isolated from pigs but cattle have been identified as the reservoir host. To date, IDV has not been confirmed to cause human disease. Like the haemagglutinin (HA) of influenza A virus (IAV) and the haemagglutinin-esterase fusion (HEF) protein of influenza C virus (ICV), the IDV HEF is produced as a precursor protein (HEF0) that must be proteolytically cleaved by host cell proteases (into HEF1 and HEF2) to gain its fusion capacity. The proteases that activate IAV HA have been extensively studied, but those responsible for activation of IDV HEF were unknown. Identifying these proteases is key to understanding early virus-host interactions and host restriction. Therefore, we generated ICV and IDV pseudotyped viruses (PVs) in HEK 293T producer cells with or without co-transfection of plasmids expressing different type II serine proteases. Subsequent transduction of swine testicular (ST) cells indicated strong activation of both ICV and IDV PVs by the human airway trypsin-like protease (HAT) and its swine homologue (swAT). Furthermore, like influenza A/Puerto Rico/8/34 (H1N1) virus, addition of exogenous protease is not essential for IDV replication in MDCK II cells, most likely due to endogenous expression of matriptase. In conclusion, our data unveil new information on host cell proteases that activate ICV and IDV HEF proteins. Importantly, the data suggest that protease specificity is not a factor in restriction of IDV replication in the human upper respiratory tract.

microbiology↗

bioRxiv2026

U-Probe: universal agentic probe design for imaging-based spatial-omics

Probe design for fluorescence in situ hybridization (FISH) underpins spatial transcriptomics, three-dimensional genome studies, and clinical diagnostics, yet remains constrained by two challenges: dependence on expert knowledge for parameter selection and quality evaluation, and the inability of existing tools to accommodate the diverse probe architectures introduced by rapidly emerging methods. Here we present U-Probe, a universal and agentic probe design platform. U-Probe employs a declarative configuration system with a directed acyclic graph (DAG)-based assembly engine that supports arbitrary probe structures from established protocols such as MERFISH, seqFISH, and MiP-seq to entirely novel architectures without code modifications. Integrated LLM-based AI agents enable conversational design workflows, allowing users to specify experimental goals in natural language and receive synthesis-ready probe sequences. We validated U-Probe in three scenarios: agent-driven MiP-seq panel design for influenza-infected mouse lung tissue, genome-tiling DNA-FISH for herpesvirus detection, and a novel RCA-based ligation probe for single-nucleotide mutation discrimination. U-Probe is available as an open-source tool with CLI, Web, and agent interfaces.

bioinformatics↗

bioRxiv2026

Space-Time Light-Sheet Microscopy

Light-sheet microscopy (LSM) has revolutionized bioimaging by delivering high-contrast volumetric resolution with minimal photodamage. Spatial wavefront shaping, used to gen{-}erate lattice and Airy light-sheets, has been particularly effective in advancing LSM be{-}yond the Rayleigh limit. Despite its broad adoption, most LSM implementations rely on rigid dual-objective geometries that complicate sample handling and impose a trade-off between imaging field of view (FoV) and axial resolution. Here, we introduce space-time light-sheet microscopy (ST-LSM), a single-objective strategy that exploits space-time (ST) correlations for the first time. ST-LSM goes beyond separate spatial or temporal modulation to jointly modulate the spatiotemporal spectral structure of a pulse. This uniquely enabled light-sheets with wavelength-scale thickness over millimeter-scale dis{-}tances. When compared to state-of-the-art approaches, ST-LSM eliminates the dual-objective constraint, expands the sample-accessible volume by 25x, and increases the FoV by 10x without sacrificing sectioning resolution. We demonstrate the versatility of ST-LSM by using a single setup to image specimens across four orders of magnitude in size, from whole roots and developing embryos, down to mammalian cells with sub-cellular axial resolution. These results position ST-LSM as an accessible and high-performance optical microscopy platform at a variety of biological scales, by translating space-time wave-packet physics into a practical imaging modality.

biophysics↗