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Li, B.

Publications and source records attributed to Li, B..

At least 127 records · Page 7Linked to original sources

scCotag: Diagonal integration of single-cell multi-omics data via prior-informed co-optimal transport and regularized barycentric mapping

Recent advances in high-throughput single-cell technologies have enabled characterization of cellular states across distinct omics layers, yielding complementary insights into the organization of biological systems. To jointly leverage these disparate modalities, computational methods have been developed to align non-overlapping cell populations profiled in different omics layers, a task known as diagonal integration, thereby facilitating more fine-grained biological interpretations. However, existing integration approaches unrealistically assume that all cells are alignable and treat prior-knowledge-derived feature correspondences as fully reliable, retaining them unrefined throughout the alignment process. In this work, we introduce scCotag, a co-optimal transport (COOT)-based deep learning frame-work for diagonal integration of single-cell multi-omics data, to address these limitations. scCotag first infers cell alignment and feature correspondence with prior-informed COOT in an iterative manner, and then leverages the resulting transport plans to jointly learn cell and feature embeddings via regularized barycentric mapping and graph reweighting. Systematic benchmarking on human brain, bone marrow, and blood single-cell RNA-seq and ATAC-seq datasets demonstrates the overall superior performance of scCotag over state-of-the-art methods in both cell alignment accuracy and embedding accuracy. In particular, scCotag excels in simulated imbalance scenarios where not all cells are alignable. Applying scCotag to data in postmortem brains of Alzheimers disease (AD) and non-AD (NoAD) patients further yields more fine-grained biological interpretation of regulatory mechanisms. Together, these results demonstrate that scCotag provides a robust framework for single-cell diagonal integration and regulatory inference.

bioinformatics↗

Mechanism of Lutein to meso-Zeaxanthin Isomerization by RPE65 Catalysis

The macular pigments lutein (L), zeaxanthin (Z), and meso-zeaxanthin (MZ) protect the human retina from light and oxidative stress. While L and Z are abundant in the human diet, MZ is nearly absent. We previously demonstrated MZ is derived from L in chicken embryos precisely timed with expression of RPE65. Herein, we show that RPE65 from mouse, an animal that does not concentrate MZ in the eye, catalyzes L to MZ isomerization similarly as found for RPE65 from chicken and human, when expressed in cultured cells. Co-expression with xanthophyll-binding proteins had no impact on MZ yield. L and MZ both fit deep into the tunnel accessing the non-heme iron center, with strain evident for the 3R,6R {varepsilon} ring of L. Interestingly, a negatively charged Glu148, found along substrate tunnel, highly conserved among carotenoid cleavage dioxygenases, and which is critical for eye health, could be replaced by a neutral, isosteric residue (Gln) without impacting MZ yield. We propose that L to MZ isomerization proceeds by a neutral, radical transition state that differs from the carbocation encountered during retinoid isomerization. These findings extend our mechanistic understanding for macular carotenoid metabolism and should be considered when developing therapeutic interventions that act via RPE65.

biochemistry↗

From features to slice: parameter-cloud modeling of spatial transcriptomics for simulation and 3D interpolatory augmentation

Computational simulation and data augmentation of spatial transcriptomics (ST) are essential for quantitative benchmarking, reproducibility, and methodological innovation. Yet current models often lack flexibility in controlling spatial and transcriptional heterogeneity, fail to capture higher-order gene dependencies, and rarely extend to three-dimensional or alignment-aware contexts. Here we present FEAST, a computational infrastructure that models ST data within a parameter cloud - a latent manifold encoding gene-level mean, variance, and sparsity. By sampling and perturbing this manifold, FEAST generates high-fidelity synthetic slices with tunable spatial and transcriptional variation, enabling systematic evaluation of clustering, deconvolution, and spatial alignment algorithms. Beyond two dimensions, FEAST performs 3D parameter-cloud interpolation guided by optimal transport and benchmark alignment, reconstructing continuous tissue architectures while preserving molecular coherence. Together, these capabilities establish FEAST as a foundational platform for standardized benchmarking, data augmentation, and 3D reconstruction in spatial transcriptomics. The source code and tutorials for FEAST are publicly available at https://github.com/maiziezhoulab/FEAST, and can be installed via Pypi at https://pypi.org/project/FEAST-py/.

bioinformatics↗

Comparative evaluation of reference genomes and cell-type annotation frameworks for single-nucleus transcriptomic analysis in apple

Single-nucleus RNA sequencing (snRNA-seq) provides a powerful tool to profile cell-type-specific transcriptional programs in woody fruit trees. Yet its application in Malus domestica has been limited by heterogeneous reference genomes and the lack of validated marker genes. Here, we first established a comprehensive set of marker genes corresponding to 29 plant cell types and identified their orthologs in apple. Using these resources, we systematically evaluated six publicly available apple genome assemblies as references for snRNA-seq analysis. Our results demonstrate that reference genome selection has a major impact on mapping performance and downstream cell type annotation, with the GDDH13 assembly consistently producing the highest transcript counts per nucleus and the greatest number of identifiable transcriptional clusters (15 clusters representing 11 distinct cell types across 35,557 high-quality nuclei). To achieve robust cell type annotation, we integrated classical marker gene-based annotation with two cross-species computational frameworks -- Orthologous Marker Gene Groups (OMGs) and XSpeciesSpanner. These three approaches enabled confident identification of 11 major cell types in apple seedlings and provided cross-validation for ambiguous or previously unannotated clusters. Pseudotime analysis reconstructed the developmental trajectories of both procambial and stomatal lineages, revealing their progressive transcriptional transitions and enabling the delineation of key molecular programs underlying vascular differentiation and early guard-cell formation. Together, this work provides a reference-quality single-nucleus transcriptomic atlas for apple and underlined the critical impact of genome choice and multi-method cell-type annotation on single-cell studies in non-model crops.

bioinformatics↗

Stimuli-Expressed Protein Qubits for Quantum Quantification of Heavy Metals in Natural Water

Environmental heavy metal pollution poses a severe threat to ecological security and human health, necessitating the development of highly sensitive, specific, and rapid detection technologies. Traditional fluorescent biosensors are often constrained by high background interference and limited sensitivity, while single-target detection systems struggle to meet the demands of complex environmental monitoring. This study constructs a novel magnetically modulated photonic quantum biosensing system based on the CadC-T7 gene circuit, replacing the traditional fluorescent reporter gene EGFP with the magnetically modulated fluorescent protein MagLOV - a quantum biomaterial. To enhance its magnetic modulation performance, the mutant with the highest lock-in amplitude during the directed evolution (R10, MagLOV 2) was selected; Simultaneously, systematic parameter optimization was conducted to determine the optimal magnetic modulation frequency (0.05 Hz), the heavy metal stimulation duration (the 2-hour group exhibited the most significant intra-group variation), and the bacterial concentration at the onset of the heavy metal stimulation (controlled at OD600 = 1 to prevent fluorescence quenching due to culture turbidity). The system demonstrates excellent multi-analyte detection capabilities for lead chloride, lead nitrate, and cadmium chloride across a concentration range of 10-8 to 10-4 M. Compared to conventional steady-state photometric methods based on EGFP and MagLOV 2, the lock-in analysis of MagLOV 2 achieves detection sensitivities enhanced by 1[~]2 orders of magnitude, with detection limits as low as 2.4 nM for cadmium chloride, 4.8 nM for lead nitrate, and 9.9 nM for lead chloride. Langmuir isotherm fitting results demonstrate high quantitative accuracy (R2 > 0.950), with particularly outstanding fitting performance for cadmium chloride (R2 = 0.999) and lead nitrate (R2 = 0.995).

synthetic biology↗

IRCAS: a novel end-to-end approach to identify, rectify and classify comprehensive alternative splicing events in a transcriptome without genome reference

Alternative splicing (AS) is a fundamental post-transcriptional mechanism that amplifies proteomic diversity and enables adaptive responses across eukaryotes. Current AS detection methods rely heavily on reference genomes, limiting their applicability to non-model organisms. Existing reference-free approaches suffer from inaccurate splice site prediction and treat detection and classification as separate processes, resulting in cascading errors. We present IRCAS, an integrated end-to-end framework for reference-free AS analysis, comprising three modules: identification, rectification, and classification. IRCAS employs colored de Bruijn graphs for AS detection, an attention-based CNN for splice site rectification, and a hybrid Graph Neural Network combining GAT and Transformer layers for classification. Evaluation across four species demonstrates substantial improvements: splice site accuracy increased to 92-96% versus 50-55% for existing methods, and end-to-end accuracy reached 83.4% compared to 41.2% for the previous best method. IRCAS establishes a new benchmark for reference-free AS detection in non-model organisms. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=167 SRC="FIGDIR/small/689457v1_fig1.gif" ALT="Figure 1"> View larger version (42K): org.highwire.dtl.DTLVardef@1162531org.highwire.dtl.DTLVardef@e3c2e2org.highwire.dtl.DTLVardef@10f9032org.highwire.dtl.DTLVardef@c3cb5e_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOFig 1.C_FLOATNO Workflow for construction and application of IRCAS. IRCAS is composed of three parts: identification, rectification, classification. (A) Workflow for reference-free AS identification from a raw transcriptomic data. First, according to the input transcripts, we apply BLAST all versus all alignment for preliminary screen. Then we adopt the MkcDBGAS Graph construction strategy. A cDBG was constructed from two sequences using a specified k-mer size. Based on bubble topologies, bubbles were classified into 5 types: SNV-induced, four AS-induced, MX-induced, AL-induced, AF-induced. (B)Workflow for AS position offset rectification and reconstruction of cDBG. Input transcript pairs are converted into a single sequence that includes two virtual nucleotides denoting the splicing start and end sites. SUPPA, a reference-based method, is utilized to determine the true splicing sites. The sequence is encoded into an nx6 vector using one-hot encoding. The offset between the true and predicted splicing sites is calculated and encoded as the ground truth. An attention-based convolutional neural network (CNN) rectification model is trained on these data to predict the offset, enabling the reconstruction of the cDBG with corrected splicing positions. (C) Workflow for 4 types AS classification. For each cDBG, node features, edge features, and global features are extracted. These features are integrated into distinct layers of a graph attention network (GAT)-Transformer hybrid model. This architecture enables high-precision classification of four types of AS events. (D)Workflow for end-to-end application of IRCAS. Transcriptomic data from any species lacking a reference genome is processed by IRCAS, enabling the classification of seven AS types with high accuracy. C_FIG

bioinformatics↗

A Syngeneic Immunocompetent Mouse Model of Gallbladder Cancer Reveals Tumorigenesis and Therapeutic Response

PurposeGallbladder cancer (GBC) is a rare but highly lethal malignancy with one of the poorest prognoses among cancers of the digestive system cancers. However, current experimental models of GBC face critical limitations, particularly their dependence on immunodeficient hosts, which precludes the investigation of the tumor microenvironment. We aimed to establish a novel syngeneic immunocompetent mouse model that recapitulates human GBC and enables the investigation of tumor-immune interactions. Experimental DesignWe engineered a murine cell line (mGBC1-ZH) from normal mouse gallbladder organoids expressing Kras and Trp53 (encoding mouse p53) mutations. Tumorigenic potential of mGBC1-ZH was evaluated by subcutaneous and orthotopic implantation. RNA-Seq and WES was used to demonstrate its characterization and similarity with human GBC. Immunohistochemistry, CCK8, and transwell assays wereused to investigate the role of CXCL5 in GBC. Therapeutic responses to standard first-line chemotherapeutic agents was evaluated in the syngeneic GBC model. ResultsThis model supports both subcutaneous and orthotopic tumor growth in immunocompetent C57BL/6J hosts while preserving hallmark features of human GBC, including biliary epithelial differentiation (CK7+/CK19+), aggressive histopathology, and an immunosuppressive "cold" tumor microenvironment. Genomic characterization revealed recurrent chromosomal instability and copy number alterations mirroring human GBC. Comprehensive transcriptomic profiling revealed profound gene expression changes during the model development, and resembled transcriptional features between mGBC1-ZH and human GBC cell lines and samples. CXCL5 was found to be upregulated in human GBC, and promote proliferation, metastasis, and neutrophils infiltration of GBC in the syngeneic mouse model. Functional validation of the syngeneic GBC model demonstrated therapeutic responsiveness to frontline chemotherapeutics gemcitabine and cisplatin with significant in vivo tumor regression. ConclusionsIn summary, we establised a novel syngeneic GBC mouse model, overcoming the limitations of traditional models by enabling studies in immunocompetent hosts. This model provides valuable insights into the molecular evolution from normal gallbladder cells to transformed cancer cells and establishes a robust platform for both mechanistic studies and therapeutic development.

cancer biology↗

Mitochondria redistribution organizes the immunosuppressive tumor ecosystem

Hostile conditions in the tumor microenvironment restrict cellular respiration, yet mitochondrial metabolism remains indispensable for tumor growth and the activity of immunosuppressive cells. How tumor ecosystems sustain mitochondrial output has been unclear. Here, we show that cancer cells resolve this paradox by acting as hubs of intercellular mitochondrial redistribution. Using mitochondrial reporter systems, we demonstrate that cancer cells import host-derived mitochondria, integrate them into their endogenous network, and subsequently relay these hybrid organelles to neighboring immune cells. Mitochondria redistribution reprograms recipient neutrophils, macrophages, and CD4+ T cells into highly suppressive states but drives CD8+ T cell exhaustion. Within cancer cells, fusion of incoming mitochondria induces filamentous P5CS assembly, enhances biosynthetic output, and enables the refurbishment of damaged organelles into fully functional units. Disrupting mitochondrial redistribution collapses the immunosuppressive ecosystem and impairs tumor growth. Thus, cancer cells do not hoard resources but orchestrate a redistribution program that fortifies their own metabolic resilience, derails anti-tumor immunity, and sustains immunosuppressive partners. HIGHLIGHTSO_LITumor cells regulate their ecosystem by redistributing mitochondria C_LIO_LIRedistributed mitochondria expand immunosuppressive cells but exhausts CD8+ T cells C_LIO_LIMitochondria fusion within cancer cells, which precedes redistribution, optimizes metabolic output by triggering conformational changes in P5CS C_LIO_LIMitochondria fusion allows cancer cells to incorporate and refurbish seemingly incompetent host-derived mitochondria, improving efficiency in the tumor ecosystem C_LI

cancer biology↗

Neurogenesis Leads Early Development in Zebrafish

Vertebrate early neurogenesis is a highly conserved process fundamental to brain function and the emergence of intelligence. However, the cellular dynamics bridging gastrulation and organogenesis remain elusive due to observational challenges. We developed a live-cell imaging platform for transgenic zebrafish that provides, for the first time, a continuous reconstruction of early neurogenesis across subcellular to organismal scales. Our analysis reveals that neurogenesis is a precisely orchestrated process. Neuronal cell bodies initially coalesce into discrete, linearly arranged clusters extending from the brain along the spinal cord. From these hubs, axons radiate outward to innervate the central nervous system and peripheral tissues, including the yolk sac surface. A primary pioneer neuron projects from the brain, coursing ventrally in parallel to the body axis. Secondary neurons then interconnect, forming a pervasive network that is subsequently refined through selective axonal apoptosis. The emergence of frequent Ca{superscript 2}{square} flashes only after structural maturation indicates that functionality is contingent upon an established scaffold. We also observe concurrent material transport and a slow, directional flow of Ca{superscript 2}{square} along axons, suggesting complementary signaling modalities. Furthermore, neurogenesis exhibits precise spatiotemporal coupling with histogenesis, particularly with the developing lateral line and vasculature. Our work, with refined spatial and time resolution, defines the kinetic pathway of early neurogenesis and underscores the critical interplay of subsystems in embryogenesis, offering fundamental insights for neural health and bio-inspired intelligence.

neuroscience↗

MERFISH+, a large-scale, multi-omics spatial technology resolves the molecular holograms of the 3D human developing heart

Hybridization-based spatial transcriptomics technologies have advanced our ability to map cellular and subcellular organization in complex tissues. However, existing methods remain constrained in gene coverage, multimodal compatibility, and scalability. Here, we present MERFISH+, an enhanced version of Multiplexed Error-Robust Fluorescence in Situ Hybridization (MERFISH), which integrates chemical probe anchoring in protective hydrogels with high-throughput microfluidics and microscopy. This optimized design supports robust and repeated hybridization cycles across an entire centimeter-scale tissue sample. MERFISH+ allowed to simultaneously quantify over 1,800 genes and resolve the 3D organization of chromatin loci and their associated epigenomic marks in developing human hearts. Using a generative integration framework for spatial multimodal data (Spateo-VI), we harmonized these MERFISH+ transcriptomic and chromatin data to reconstruct a 3D spatially-resolved multi-omic atlas of the developing human heart at subcellular resolution capturing 3.1 million cells across 34 distinct populations. This 3D atlas provides a holistic view of an entire organ enabling the characterization of 3D cellular neighborhoods and transcriptional gradients of substructures such as the descending arteries. Thus, MERFISH+ offers a robust, large-format platform for spatial multi-omics that enables high resolution mapping of gene expression at subcellular resolution and the characterization of cellular organization within 3D organs. One Sentence SummaryMERFISH+ is an spatial multi-omics platform that integrates hydrogel-based probe anchoring, automated high-throughput microfluidics, and large-format multimodal data production to enable comprehensive, subcellular resolution mapping of gene expression and chromatin organization across millions of cells within complex developing human organs. HighlightsO_LIMERFISH+ expands MERFISH capabilities to measure >1,800 genes and at whole-organ 3D imaging scale C_LIO_LICombines chemical probe anchoring with high-throughput volumetric microscopy and microfluidics C_LIO_LIGenerates a 3D molecular atlas of a developing human heart with > 3.1 million cells at subcellular resolution C_LIO_LIIntroduces Spateo-VI, a novel generative framework integrating 3D multimodal datasets C_LI

genomics↗

Signal peptidase complex mediates rotavirus VP7 processing and virion assembly

For viruses that replicate in the proximity of or bud at the endoplasmic reticulum (ER) associated membranes, proper processing of their glycoproteins is critical for successful infection. Rotavirus outer capsid protein VP7 is an ER-resident protein. However, its N-terminal signal peptide is removed by an unknown proteolytic mechanism. In this study, we leveraged tandem affinity purification followed by high-resolution mass spectrometry to profile host proteins that interact with VP7. We identified members of the signal peptidase complex (SPC) family as important host factors that facilitate rotavirus infection. CRISPR knockout or siRNA knockdown of distinct SPC subunits resulted in significant decrease in infectious rotavirus titers in a viral strain- and cell type-independent manner. While viral transcription, translation, and replication were not altered in the absence of SPC, we observed formation of abnormal viral particles by transmission electron microscopy (TEM) in SPCS1 knockout cells. Mechanistically, loss of SPC proteins led to inefficient cleavage of VP7 signal peptide and severely impaired the final steps of virion maturation and assembly. Additionally, we identified residue E256 within VP7 as a key site for SPC binding. An E to R mutation abolished VP7 interaction with SPC and subsequently led to reduced viral infectivity. Taken together, these findings define SPC as a novel regulator of VP7 maturation and rotavirus assembly and highlight its role as a novel cellular target for potentially broad-spectrum antiviral therapeutic development. Author summaryPrevious reports demonstrated that SPC is involved in the replication cycles of several members of the flavivirus family and human T-cell leukemia virus type 1 (HTLV-1). As an ER-resident outer capsid protein, rotavirus VP7 must undergo proper post-translational modifications, including the cleavage of its N-terminal signal peptide, to be functionally incorporated into mature virions. However, the processing mechanism remains unknown. For the first time, our study identifies SPC as an essential regulator of rotavirus assembly by mediating the cleavage of the VP7 signal peptide. Loss of SPC impairs VP7 signal peptide cleavage and maturation, thereby disrupting correct virion assembly and reducing infectious particle production. Using Alphafold3, we predicted the VP7 residue E256 to be at the binding interface with SPC complex. Experimentally, mutation of glutamic acid to arginine (E256R) substantially weakens this interaction and results in reduced viral propagation. Our findings unveil a novel post-translational checkpoint in rotavirus regulated by SPC and underscore the promise of SPC as a broad-spectrum antiviral target, especially for rotavirus, flavivirus, and HTLV-1, whose viral glycoproteins and structural proteins require SPC processing for proper maturation.

microbiology↗

TOMM40 suppression promotes neuronal cholesterol imbalance and molecular and behavioral phenotypes of Alzheimer's disease

INTRODUCTIONWhile the APOE4 allele is a major risk factor for Alzheimers disease (AD), the role of TOMM40--an adjacent gene involved in mitochondrial protein import--is not known. METHODSMice, human iPSC-derived neurons (iNeurons), and human brain tissue were used for study of animal cognition, cholesterol metabolism, mitochondrial function, and gene expression. RESULTSTOMM40 knockdown (KD) impaired memory in mice and increased cholesterol and A{beta} 42 in mouse brains and human iNeurons. KD disrupted mitochondria-endoplasmic reticulum contact sites (MERCs), causing mitochondrial dysfunction and promoting reactive oxygen species that led to activation of LXRB (NR1H2), upregulation of APOE and LDLR. and increased cellular cholesterol and A{beta} 42 independent of APOE4. Human brain transcriptomics showed reduced TOMM40 expression that correlated with cholesterol regulatory gene expression, amyloid burden, and clinical AD diagnosis. DISCUSSIONTOMM40 is a novel mediator of AD pathology through dual effects on MERCs that regulate cholesterol homeostasis and mitochondrial function. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=163 SRC="FIGDIR/small/685963v1_ufig1.gif" ALT="Figure 1"> View larger version (31K): org.highwire.dtl.DTLVardef@e3e3e2org.highwire.dtl.DTLVardef@1861f67org.highwire.dtl.DTLVardef@77a813org.highwire.dtl.DTLVardef@1bb9a83_HPS_FORMAT_FIGEXP M_FIG C_FIG

molecular biology↗

Distinct and compensatory roles of Stag1 and Stag2 in post-mitotic genome refolding

The three-dimensional architecture of the eukaryotic genome is largely shaped by the cohesin complex, which contains either Stag1 or Stag2 subunits. Although both subunits contribute to chromatin organization, their specific functions in de novo loop formation during post-mitotic genome refolding remain elusive. Here, we leverage the mitosis-to-G1 transition to dissect their individual roles. We found that Stag1 depletion has a negligible impact on post-mitotic genome restructuring or transcription reactivation. In contrast, Stag2 orchestrates chromatin remodeling in a manner that is both cell cycle stage-specific and chromatin context-dependent. During early-G1, Stag2 preferentially associates with euchromatin, where it drives the rapid formation of small chromatin loops. This facilitates prompt promoter-enhancer (P-E) contact formation and enables efficient transcription activation. As the nuclear concentration of Stag2 increases by late-G1, it progressively suppresses large loops, likely due to its shorter chromatin residence time and its potential to competitively displace the more extrusion-capable Stag1-associated cohesin. Mechanistically, Stag2-mediated loop extrusion is constrained by CTCF-bound barriers, rather than by genomic travel distance. Although Stag2 associates rapidly with euchromatin in early-G1, its recruitment to heterochromatin is delayed until late-G1. Simultaneous depletion of both Stag proteins results in a synergistic loss of virtually all structural loops and a more severe disruption of transcription than that caused by individual deletions. Together, these results establish Stag2 as the principal regulator of post-mitotic genome reorganization among Stag paralogs, mediating spatiotemporal control of chromatin architecture, while Stag1 provides compensatory support to ensure functional robustness.

molecular biology↗

Expression and localization of NMDA receptor GluN2 subunits in dorsal horn pain circuits across sex, species, and late postnatal development

Despite being essential mediators of pain processing, the molecular identity of N-methyl-D-aspartate receptor (NMDAR) subtypes in nociceptive dorsal horn circuits is poorly understood, especially between sexes and in humans. Given the importance of GluN2 subunits in shaping NMDAR function and plasticity, we investigated the expression and localization of specific GluN2 NMDAR variants in the dorsal horn of viable spinal cord tissue from male and female rodents and human organ donors. Analysis of single-cell/nuclei sequencing datasets and quantitative reverse transcriptase polymerase chain reactions (qRT-PCR) revealed that the GluN2A (GRIN2A) and GluN2B (GRIN2B) subunits are robustly expressed in dorsal horn neurons of mice, rats and humans, with moderate expression of GluN2D (GRIN2D). Immunohistochemistry (IHC) with antigen retrieval demonstrated that GluN2A, GluN2B, and GluN2D proteins are all preferentially localized to the superficial dorsal horn of both adult rats and humans, which is conserved between males and females. Surprisingly, we found that these GluN2 NMDAR subunits are enriched in the lateral superficial dorsal horn in rats but not in humans, while presynaptic and neuronal markers are symmetrically distributed across the rat mediolateral axis. A dramatic shift in localization of GluN2A to the lateral superficial dorsal horn was observed across later postnatal development (PD21-PD90) in both male and female rats, with a corresponding change in synaptic NMDAR currents. This discovery of changes in NMDAR subunit distribution during maturation and between species will shed light on the physiological roles of NMDARs and their potential as therapeutic targets for pain. SIGNIFICANCE STATEMENTWe used complementary single-cell/nuclei analysis, immunostaining, quantitative reverse transcriptase polymerase chain reactions, RNAscope in situ hybridization, and electrophysiological approaches to compare the relative expression of N-methyl-D-aspartate receptor (NMDAR) GluN2 subunits in dorsal horn spinal cord pain circuits of mouse, rat, and human spinal cord tissue. Through these comparisons, we find that the transcripts and proteins of the GluN2A, GluN2B, and GluN2D NMDAR subunits are robustly expressed in superficial dorsal horn neurons, with conserved expression across sex but important differences in expression and localization patterns across late development and between species. These discoveries shed light on the physiological roles of NMDARs and their utility as potential therapeutic targets for pain.

neuroscience↗

Fluke-borne viruses are a risk factor for diseases caused by carcinogenic trematodes

Three trematode parasites, Schistosoma haematobium, Opisthorchis viverrini, and Clonorchis sinensis are recognized group 1 carcinogens for their roles in squamous cell carcinoma of the urinary bladder and bile duct cancer (cholangiocarcinoma). We aimed to assess the presence of fluke viruses and their association with malignancy in fluke infections. We showed that all three carcinogenic trematodes harbored related viruses. Focusing on O. viverrini, we observed virus persistence at discrete life stages and viral tropism for the fluke tegument, and we provided molecular and serological evidence of parasitized hosts exposure to viral RNA and proteins. Serological analyses confirmed seroconversion of residents in endemic sites and revealed that antibody titers and avidity against fluke-borne viruses increase in individuals with hepatobiliary diseases. Random forest models discriminated efficiently between liver-fluke-infected participants without disease and those with periductal fibrosis or cholangiocarcinoma. Thus, we show that fluke-borne viruses are risk factors and diagnostic targets for fluke-associated cancer.

microbiology↗

Insights into Temporal and Spatial Dynamics of Short Association Fiber Formation in the Human Fetal Brain

Short association fibers (SAFs) form the local scaffold of cortical connectivity, supporting early functional specialization and marking sites of neurodevelopmental vulnerability. However, their development before birth remains largely unknown. Leveraging advanced fetal diffusion MRI and a histology-validated framework, we present the first in-utero reconstruction of SAF pathways in the human brain. We tracked their volumetric and microstructural developmental trajectories in 243 fetuses spanning a critical period when the brains connectome is rapidly forming. We found that SAFs emerge before sulcal folding, initially as flat, loosely arranged pathways along the subplate-white matter interface, and later reorganize into coherent U-shaped bundles. Their maturation followed a sensorimotor-to-association gradient, paralleling cortical development. Nonlinear, bundle-specific trajectories captured multiphasic maturation, with subplate dynamics preceding increases in axonal coherence and early myelination. By filling a missing link in lifespan brain connectivity, this study establishes a prenatal reference for cortical wiring and provides tools for investigating the origins of neurodevelopmental disorders.

neuroscience↗

ACE2 Phosphorylation Modulates Angiogenesis via the Activator Protein-1

BackgroundAngiogenesis plays a crucial role in organ development. However, aberrant blood vessel growth is involved in various diseases, including tumors and neovascular eye diseases. Angiotensin-converting enzyme 2 (ACE2) is a critical enzyme regulating the health of cardiovascular system, and its post-translational modifications (PTMs) are crucial to determine ACE2 expression level and activity. Here, we studied how the PTM of ACE2 in vascular endothelial cells (ECs) affect pathological retinal neovascularization and tumor angiogenesis. MethodsThe angiogenic capabilities of ECs were assessed by tube formation, sprouting assays, and 5-ethynyl-2-deoxyuridine (EdU) and filopodia staining. EC angiogenesis was examined by poteome profiler array and aortic ring assays in vitro and by the oxygen-induced retinopathy (OIR) and tumor angiogenesis models in vivo. High-throughput screening involving data from RNA-seq, ATAC-seq, and ChIP-seq were used to explore the epigenetic and transcriptional regulations of pro-angiogenic genes regualtged by ACE2 PTMs. ResultsACE2 Ser-680 dephosphorylation in connection with Lys-788 ubiquitination increased EC angiogenic phenotype, which were manifested by aberrant vascularization in mouse OIR and tumor models. ACE2 Ser-680 dephosphorylation led to the activation of activator protein-1 (AP-1), which transactivated multiple genes involved in angiogenesis. AP- 1 inhibition mitigated such angiogenesis in vivo. ConclusionOur findings show a novel PTM mechanism of ACE2 involved in pathological angiogeneis. Specifically, ACE2 Ser-680 dephosphorylation facilitated AP-1 transactivation of the downstream pro-angiogenic genes in ECs.

cell biology↗

Uterine stromal Erbb3-Igf1 signaling is critical to functional gland development conducive to implantation

Preimplantation uterine gland differentiation, involving glandular branching, is a prerequisite for secretion of leukemia inhibitory factor (LIF), thereby establishing uterine receptivity to embryo implantation. This differentiation process is disrupted by the absence of FOXA2, a transcription factor specifically present in uterine glands. It remains unclear whether preimplantation glandular differentiation is regulated exclusively by gland-intrinsic mechanisms or is influenced by the surrounding microenvironment through mesenchymal-epithelial interactions. We show here that uterine deletion of Erbb3 leads to impaired gland branching and FOXA2 expression, as well as reduced female fertility. In contrast, females with uterine epithelial-specific deletion of Erbb3 show normal fertility, suggesting a critical role of stromal ERBB3 signaling. A transcriptomic profiling of day 3 stromal cells identified Igf1 as a candidate paracrine factor mediating the stromal-epithelial interaction. Thus, the uterine deletion of Igf1r leads to a reduced gland branching phenotype similar to that observed in Erbb3d/d uteri. These results demonstrate that ERBB3 regulates glandular branching and embryo implantation via the ERBB3-IGF1-IGF1R signaling pathway.

developmental biology↗