Biology subjects

Yang, Q.

Publications and source records attributed to Yang, Q..

At least 37 records · Page 2Linked to original sources

bioRxiv2026

NFYA regulates spermatogenesis through multifaceted chromatin interactions

During spermatogenesis, germ cells undergo extensive chromatin remodeling to promote highly diverse and stage-specific transcriptional programs that drive differentiation, meiosis, and spermiogenesis. However, conventional loss-of-function approaches do not allow systematic interrogation of gene function across spermatogenesis in vivo. Here, we introduce a protein-degron strategy coupled with intra-seminiferous tubule injection (dTAG-IS) to enable rapid, simultaneous, and stage-resolved functional investigation of NFYA during spermatogenesis. Acute NFYA-depletion induces apoptosis in differentiating spermatogonia (diff-Spg), pachytene spermatocytes (pSpc), and round spermatids (rStd), revealing an essential requirement across spermatogenesis. Mechanistically, NFYA exhibits stage-specific chromatin-binding patterns and directly regulates distinct transcriptional programs, including cell cycle progression and histone assembly in diff-Spg; chromosome organization and DNA damage response in pSpc; and antioxidative stress programs in rStd. Together, our study underscores dTAG-IS as a versatile strategy for in vivo loss-of-function studies during spermatogenesis and establishes NFYA as a multifaceted regulator that drives stage-specific transcriptional programs to ensure spermatogenesis progression.

developmental biology↗

bioRxiv2026

Medulloblastoma Forms Symbiotic Metabolic Partnerships with Macrophages to Establish Leptomeningeal Metastases

Leptomeningeal metastases are the primary source of morbidity and mortality for pediatric medulloblastoma patients. Due to limited surgical sampling of metastases in patients, little is understood of the mechanisms of metastasis. Here, we identify biologically distinct quiescent small metastases (designated as micrometastases) and mitotically active larger metastases (macrometastases). Macrometastases are more metabolically active than micrometastases and contain higher levels of lipids, particularly cholesterol. Macrometastases secrete CXCL12, which attracts lipid-laden macrophages into the tumor. Lipid-laden macrophages upregulate the cholesterol transporter ABCG1, promoting the efflux of free cholesterol, which is then taken up by tumor cells via the HDL receptor SCARB1. CXCL12-driven macrophage recruitment and exogenous cholesterol are sufficient and necessary to drive progression of medulloblastoma leptomeningeal metastases in vivo. High fat diets drive metastatic progression in vivo. Dietary or pharmacological interventions targeting the CXCL12-SCARB1-cholesterol axis represent therapeutic strategies to either prevent or treat medulloblastoma leptomeningeal metastases.

cancer biology↗

bioRxiv2026

Deciphering complete archaic introgression sequences in modern human genomes

Genetic introgression from archaic hominins has profoundly reshaped the genetic diversity and adaptive potential of modern humans, yet the full catalog of introgressed sequences, particularly those residing in structurally complex regions has remained elusive. Here, we present ASMaid (ASseMbly-based archaic introgression detector), a Hidden Markov Model-based framework that leverages haplotype-resolved pangenome assemblies to identify archaic-derived sequences with unprecedented completeness. By integrating both single-nucleotide genotype and structural variation (SV) signals, ASMaid captures significantly more intact archaic segments than conventional reference-based approaches. Applying ASMaid to a global panel of 610 phased human genome assemblies, we show that non-African individuals carry approximately 79.8 Mbp of Neanderthal and 8.3 Mbp of Denisovan sequences, representing substantial increases over previous estimates, respectively. Notably, we detected several centromere-spanning archaic segments, including EAS-specific calls on chromosomes 5 and 7. Our assembly-based approach uncovered 1,701 archaic-derived SVs, revealing a previously overlooked layer of archaic functional legacy. High-frequency introgressed loci are enriched in pathways associated with metabolism, immunity, and nervous system (e.g. CTNNA2 linked to early-onset schizophrenia risk), underscoring the fundamental role of introgression in modulating modern human traits. Notably, we identified dozens of loci potentially facilitating local adaptation, such as PRDM16 involved in adipocyte differentiation and cold tolerance, and CSGALNACT2 associated with chondroitin sulfate synthesis. Furthermore, our analysis delineates three distinct Denisovan introgression pulses in Eastern Eurasian genomes, in which the first two pulses are shared across East Eurasian and Oceanian populations, while the third remain primarily exclusive in East Asians. Reflecting these complex introgression events, 31 Denisovan-derived segments, including the TBX15-WARS2 locus, are inferred to have been introduced via at least two events. This comprehensive map of archaic introgression provides a fundamental resource for understanding how ancient gene flow continuously shapes human phenotypic diversity and adaptation.

evolutionary biology↗

bioRxiv2026

Peripheral sensory terminal degeneration is linked to sensory fiber hyperexcitability in paclitaxel-induced peripheral neuropathy

Paclitaxel-induced peripheral neuropathy (PIPN) is the most common dose-limiting side effect of paclitaxel chemotherapy, yet the relationship between structural damage to peripheral nerve terminals and functional impairment of nociceptors has not been directly examined. Here we combined ex vivo skin-nerve electrophysiology with three-dimensional (3D) imaging of fDISCO-cleared glabrous skin to characterize both morphological and functional changes in peripheral sensory terminals following paclitaxel treatment in rats. Five weeks after paclitaxel administration, skin-nerve recordings revealed a marked increase in the proportion of C- and A{delta}-fibers exhibiting spontaneous discharge (approximately 2.4- and 2.6-fold increases, respectively) compared to vehicle-treated controls. Paclitaxel also selectively reduced the mechanical activation threshold of C-fibers without affecting A{delta} fibers. Three-dimensional reconstruction of PGP9.5-immunolabeled nerve terminals showed that sensory fibers in glabrous skin form a vertically oriented, tree-like architecture. Paclitaxel treatment severely reduced both the terminal branch length and the density of free nerve endings in the epidermis. Strikingly, co-administration of the Kv7 channel activator retigabine prevented both the electrophysiological and morphological alterations induced by paclitaxel. These findings provide direct evidence that peripheral nerve terminal degeneration and hyperexcitability co-occur in PIPN and that Kv7 channel activation can protect against both structural and functional damage. SignificanceThis study provides the first direct evidence linking intraepidermal sensory terminal degeneration with peripheral sensory fiber hyperexcitability in paclitaxel-induced peripheral neuropathy by combining skin-nerve electrophysiology with three-dimensional imaging of sensory terminals. Paclitaxel induces distal degeneration of intraepidermal sensory terminals, increases spontaneous discharge, and lowers the mechanical activation threshold of C-fibers, demonstrating that structural degeneration and functional abnormalities occur concurrently in the peripheral terminal. Retigabine prevents both structural and functional alterations, supporting peripheral sensory terminals as a therapeutic target for preventing chemotherapy-induced neuropathy.

neuroscience↗

bioRxiv2026

An early CYP26A1/CRYAA progenitor-glial domain marks the presumptive macular region during human retinal development

The human macula, a high-acuity retinal region essential for central vision, emerges early in development, but the cellular basis of its regional specialisation remains incompletely resolved. Building on recent studies that identified CYP26A1-mediated retinoic acid (RA) suppression in the presumptive macula, we define a CRYAA-positive progenitor-glial compartment in the temporal human retina from post-conception week (pcw) 7 onward. Single-cell RNA sequencing, immunohistochemistry and spatial morphometry indicate that this compartment is linked to regional gliogenic maturation and later corresponds to a specialised macular Muller glial population, rather than simply reflecting uniform pan-retinal developmental timing. Spatial mapping shows that the CYP26A1-positive domain remains delimited as the surrounding retina expands. Adult tissue analysis shows that CYP26A1 expression is retained in primate macular Muller glia and in corresponding acute-zone regions of visual-streak species. These findings extend current models of CYP26A1-mediated RA modulation in human acute zone/macular development by linking the early CYP26A1 domain to a regionally specialised CRYAA-positive progenitor-glial compartment. We propose that early macular patterning includes a spatially restricted glial programme that may contribute to later regional architecture and disease vulnerability.

developmental biology↗

bioRxiv2026

A novel AR molecular glue degrader via bivalent engagement of the N-terminal domain and RNF213-UBE2J2 complex

Targeted protein degradation enables the elimination of disease-relevant proteins through induced proximity between E3 ligases and substrates. Current androgen receptor (AR) therapies act via the ligand-binding domain (LBD) but fail against constitutively active LBD mutants or splice variants such as AR-V7. Here, we identify GZL626, a small-molecule degrader that induces proteasome-dependent degradation of both full-length AR and AR-V7. GZL626 suppresses AR transcriptional activity, inhibits proliferation of AR-positive prostate cancer cells, and blocks SARS-CoV-2 infection. Mechanistically, GZL626 binds the intrinsically disordered AR N-terminal domain (NTD) and promotes formation of a noncanonical RNF213-UBE2J2-AR ternary complex that drives AR ubiquitination and degradation. The AR DNA-binding domain (DBD) acts as a structural hub within this assembly. Distinct from PROTACs or classical molecular glues, GZL626 engages both AR and RNF213 bivalently, remodeling protein interactions to stabilize a functional E3-E2-substrate complex. These findings establish GZL626 as a first-in-class NTD degrader with potential to overcome resistance in advanced prostate cancer.

biophysics↗

bioRxiv2026

Dynamic resource allocation orchestrates physical simulation in the human brain

Humans can flexibly predict physical events by internally simulating object dynamics in the brain--a capacity lacking in current AI systems. Using a ball collision paradigm with visual occlusion combined with multimodal neuroimaging (fMRI/MEG), we uncover a spatiotemporally organized neural architecture for physical simulation. fMRI reveals hierarchical spatial segregation: higher-order cortical regions encode relational physical variables, distinct from object-specific features encoded in the lower-order sensorimotor regions. MEG uncovers two temporally distinct neural processes: a real-time simulation tracking the evolving state of the object, in alignment with collision dynamics, and an early predictive signal anticipating collision occurrence ([~]700 ms before contact). We propose that this early predictive control mechanism dynamically allocates cognitive resources during simulation. This is formalized by a Dynamic Resource Intuitive Physics Engine (IPE) model, which captures both behavioral data and the dual neural timescales by optimizing accuracy-cost tradeoffs. Crucially, this framework predicts early encoding of another adaptive control variable separate from collision occurrence, as evidenced by both MEG and pupillary responses. These findings reveal how the brain achieves efficient physical inference through hierarchically organized predictive control that dynamically allocates cognitive resources.

neuroscience↗

bioRxiv2026

A Polymeric Nanoparticle System for the Delivery of CRISPR/Cas9 Components into Arabidopsis Pollen

Efficient and heritable genome editing in plants remains constrained by transformation bottlenecks and reliance on tissue culture-based regeneration. Targeting the male germline offers a promising alternative for DNA-free genome modification. Here, we report a polymeric nanoparticle platform for the delivery of RNA-based CRISPR/Cas9 components into Arabidopsis thaliana pollen, establishing a foundation for sperm transfection-assisted genome editing (STAGE)-like approaches in plants. Poly(2-dimethylaminoethyl methacrylate) (PDMAEMA)-based polyplexes were designed to independently encapsulate Cas9 mRNA and ATTO 550-labeled guide RNA, forming nanoparticles with hydrodynamic diameters of [~]146 nm and condensed cores of 20-30 nm. Following internalization and cytosolic release, Cas9 mRNA translation enabled the nuclear localization of ATTO 550-labeled gRNA, as confirmed by confocal imaging and fluorescence lifetime (TauSense) analysis. Fluorescent signals corresponding to the CRISPR RNPs were detected in both vegetative and sperm cell nuclei, with higher accumulation in the vegetative nucleus. Together, these results demonstrate the feasibility of RNA-mediated delivery and intracellular assembly of CRISPR/Cas9 RNPs in plant male gametophytes. By bypassing tissue culture and DNA integration, this nanoparticle-based approach establishes a framework for a more efficient mechanism for introducing heritable genome modifications in plants. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=91 SRC="FIGDIR/small/733037v1_ufig1.gif" ALT="Figure 1"> View larger version (51K): org.highwire.dtl.DTLVardef@170f7a5org.highwire.dtl.DTLVardef@1929c6forg.highwire.dtl.DTLVardef@5c4a94org.highwire.dtl.DTLVardef@1243cc8_HPS_FORMAT_FIGEXP M_FIG C_FIG

Plant Biology↗

bioRxiv2026

Structural Basis of Nidovirus Replication Organelle Evolution Revealed by the Arterivirus DMV Pore Complex

Positive-strand RNA viruses of the order Nidovirales - which include coronaviruses and arteriviruses - remodel host membranes into double-membrane vesicles (DMVs) as replication organelle to shield viral RNA synthesis from immune sensors. In coronaviruses, newly synthesized RNA is exported to the cytoplasm through a massive DMV pore complex formed by viral non-structural proteins (nsps). However, it remains unknown whether this elaborate architecture is unique to large-genome coronaviruses or a universal hallmark of the order. Here, we integrate in situ cryo-electron tomography and single-particle cryo-electron microscopy to resolve the atomic structure of DMV pore complex within its native membrane environment, from Equine Arteritis Virus (EAV), a prototype arterivirus with small genome. Despite lacking obvious sequence homology, the minimal EAV pore shares conserved architectural principles with its elaborate coronavirus counterpart. EAV pore complex is formed by a 12:12 stoichiometry of nsp2 and nsp3 protomers organized into four stacked concentric rings on double-membrane, yet generating pronounced structure symmetry mismatch. Functionally, the complex displays a distinct pore profile while preserving a positively charged central channel, essential for viral replication and transport. These findings demonstrate that, despite diversity in genome size and virion morphology, nidovirus replication organelles exhibit striking evolutionary conservation at the atomic structural level. Collectively, we propose that the order Nidovirales can be unified at the ultrastructural level by this conserved signature pore complex on the DMV-based replication organelle.

microbiology↗

bioRxiv2026

Structural basis of substrate recognition and allosteric inhibition in human B0AT2

The SLC6 family is a major target for neuropsychiatric therapeutics. Human B0AT2 (SLC6A15) regulates amino acid homeostasis and glutamatergic transmission and is linked to major depressive disorder, yet its transport and inhibition mechanisms remain unclear. Here we report cryo-EM structures of B0AT2 in apo state and in complex with substrates (proline, leucine, methionine) and inhibitors (loratadine, tiagabine), capturing outward-open, early substrate-bound intermediate, outward-occluded, and inward-open conformations along the transport cycle. These structures reveal an early substrate-sensing mechanism at the substrate-binding pocket (S1), where a substrate-dependent rotameric switch of Phe308 remodels pocket geometry to tune substrate accommodation and selectivity. Loratadine stabilizes an outward-occluded state via allosteric inhibition at the extracellular S2 pocket, whereas tiagabine stabilizes the inward-open state through cooperative multi-site inhibition involving S1 and two previously unrecognized intracellular cavities (S3 and S4). Together with functional assays, these data define the molecular basis of B0AT2 substrate selectivity and state-dependent inhibition. Notably, the two intracellular cavities are conserved across SLC6 transporters, reflecting a shared intracellular vestibular architecture and enabling rational design of conformation-selective modulators for neuropsychiatric disorders.

biophysics↗

bioRxiv2026

Low-Energy Polishing Facilitates Breaking the Resolution Barrier of In Situ Cryo-EM

In situ structural analysis allows direct visualization of protein structures in their native cellular environments, but near-atomic resolution in cellular lamellae has been significantly limited to exceptionally large complexes such as ribosomes. A key factor underlying this limitation is the degradation of data quality of thin lamellae caused by substantial subsurface damage from cryo-focused ion beam (cryo-FIB). Here, we developed a cryogenic low-energy polishing in FIB approach, which reliably produces thin lamellae with low damage across different cell types. This advance, combined with in situ single particles analysis, has pushed down the molecular weight lower limit for in situ reconstruction at near-atomic resolution to [~]400 kDa. We demonstrate this by resolving photosynthetic complexes (3.4 [A] and 3.3 [A]), metabolic enzymes (3.3 [A]), chloroplast ribosome (4.0 [A]) and respiratory chain complexes (3.7 [A]) from Chlamydomonas reinhardtii. Furthermore, the efficient workflow enables rapid structural feedback upon changes in cellular states, offering a practical way to perform multi-condition in situ structural analysis.

biophysics↗

bioRxiv2026

NEO-AAV: an engineered extracellular vesicle-enveloped AAV platform for activation-coupled T cell transduction and CAR-T cell generation

Efficient delivery of genetic cargo to primary human T cells remains a critical barrier for cell-based immunotherapies. AAVs are widely used but limited by immune neutralization and poor T cell transduction efficiency. We present NEO-AAV, an engineered fusion protein (PH-ALG2-PKD12) that recruits AAV capsids into endogenous extracellular vesicles (EVs) via PI(4,5)P2-directed membrane targeting, multivalent PKD12-capsid clustering, and ALG2-mediated ESCRT machinery recruitment, yielding EV-enveloped particles with improved AAV loading compared with passive EV-AAV controls. NEO-AAV displayed enhanced resistance to an anti-AAV6 neutralizing antibody (ADK6), maintaining transduction at concentrations that neutralized naked AAV6 and outperformed passively formed EV-AAV6. Surface display of a CD7/CD3/CD28 tri-chimera enabled single-step activation and transduction of CD7 T cells within PBMCs, reaching [~]38% eGFP cells without exogenous pre-activation. As a proof-of-concept, NEO-AAV generated functional CAR-T cells from primary human PBMCs, exhibiting antigen-specific IFN-{gamma} release and cytotoxicity against CD19+ target cells. CAR expression peaked at day 5 and declined by day 15, consistent with episomal AAV kinetics, framing NEO-AAV as an activation-coupled delivery module rather than a durable solution. Together, NEO-AAV provides a programmable EV-enveloped AAV platform with improved immune shielding and single-step T cell transduction, offering a building block for immune cell engineering with in vivo potential.

bioengineering↗

bioRxiv2026

A regulatory TRIF/IL-1R1 axis controls T-dependent IgA production in the intestines

Mucosal IgA is critical for controlling the microbiota and preventing pathogenic infection of the epithelium. The innate signals that regulate the generation of IgA remain poorly defined. Here, we identified TRIF and IL-1R1 as immune checkpoints for intestinal IgA production. In the absence of infection, Trif-/- and Il1r1-/- mice exhibited markedly elevated stool IgA and IgA-bound commensals. We show that IL-1R1 restricts IgA class-switching in a B cell-intrinsic manner, while TRIF acts extrinsically of B cells and shapes the Peyers patch microenvironment. Loss of either Trif or Il1r1 enhanced retinoic acid metabolism and allowed premature Ccnd3 upregulation in naive B cells, favoring both IgA class-switching and differentiation into germinal center B cells. During oral vaccination, the absence or blockade of the TRIF/IL-1R1 pathway increased antigen-specific IgA production without affecting seric antigen-specific IgG levels. These findings unveil novel and local signaling targets to promote robust antigen-specific mucosal immunity.

immunology↗

bioRxiv2026

Revisiting the diversity of secondary endosymbionts in the major pest oat aphid, Rhopalosiphum padi

There is disagreement on whether secondary endosymbionts are found in the major cereal pest aphid, Rhopalosiphum padi. Some papers report a diversity of secondary bacterial endosymbionts while others have failed to find evidence of these bacteria in this species. Here we revisit this issue by summarizing the relevant literature and through additional sampling of the species in Australia, China and Denmark using a combination of molecular approaches. We find a general absence of secondary endosymbionts beyond the obligate endosymbiont Hamiltonella defensa in R. padi. While the inconsistency in survey results may reflect rapid changes in endosymbiont turnover in populations and/or the impact of ecological factors such as host plant type on endosymbiont diversity, we are concerned that technical issues may be at least partly responsible for inconsistencies in the literature. This leads us to emphasize the importance of multiple sources of evidence required to establish and characterize endosymbiont infections, including PCR and qPCR assays, DNA Sanger sequencing and 16SrRNA gene metabarcoding. We note that several major aphid pests show a low incidence of secondary endosymbionts which raises issues about the importance of these endosymbionts in aphids that constitute pests, even though endosymbionts can in some cases increase host fitness and therefore pest impact.

microbiology↗

bioRxiv2026

Rhythmic temporal structure organizes recurrent dynamics to support sequential working memory

Rhythmic temporal structure improves working memory, but how this benefit emerges from recurrent dynamics remains unclear. Here, we trained excitatory-inhibitory recurrent neural networks with short-term synaptic plasticity to perform a sequential delayed match-to-sample task with either regular or jittered sample timing. Rhythmic input produced a small but reliable improvement in task accuracy and was associated with more differentiated population trajectories during encoding. This behavioral advantage was accompanied by an organization of population dynamics around the dominant input frequency: temporal regularity progressively brought stimulus arrivals closer to preferred encoding phases, modulated phase advancement during stimulus presentation, and reduced the deviation of inter-stimulus phase-progression frequency from the dominant input rhythm. As a result, internal oscillations increasingly tracked the temporal structure of the input across the sequence, providing a phase-based scaffold for encoding ordered information. This scaffold preferentially supported temporal-order representations rather than uniformly enhancing all stimulus features. Decoding analyses further showed that stronger temporal regularity increased the fidelity and persistence of stimulus information in both neuronal activity and synaptic efficacy, whereas perturbing synaptic efficacy produced the largest impairment during delay-period maintenance. These findings suggest that rhythmic input supports sequential working memory by imposing a reliable temporal structure on recurrent dynamics and stabilizing synaptic-state representations.

neuroscience↗

bioRxiv2026

DamageFormer: a damage-aware multimodal deep learning framework for DNA lesion identification from nanopore sequencing

BackgroundDNA lesions arise from endogenous metabolism and environmental exposure and are the major drivers of mutagenesis, aging, and cancer development. However, mapping DNA damage at nucleotide resolution remains a technically challenging task. Nanopore sequencing enables direct detection of chemical perturbations through alterations in ionic current signals. Despite this potential, existing computational approaches remain limited in their capacity to generalize across diverse lesion types and to effectively integrate nucleotide sequence context with raw signal information for accurate detection and localization. ResultsWe presented DamageFormer, a multimodal deep learning framework for detection and localization of DNA lesions using native nanopore sequencing data. Central to this framework is LesionBERT, a damage-aware genomic foundation model built upon DNABERT-2 and enhanced with lesion-focused reconstruction objectives to improve representation of chemically modified bases. DamageFormer integrated LesionBERT with a neural signal model through an adaptive gating mechanism, enabling dynamic weighting of sequence context and nanopore signal evidence. The model was trained using a joint objective that combines prediction, localization, and contrastive alignment losses to promote cross-modal coherence and spatial precision. On an oxidative DNA damage benchmark comprising paired sequence and signal data, DamageFormer achieved an AUROC of 0.99997 for lesion detection and a mean absolute localization error of 0.00439, consistently outperforming state-of-the-art baselines. Model interpretation analyses revealed context-dependent modality weighting that adapts to variation in signal quality and sequence ambiguity. The proposed framework further generalizes to chemically distinct guanine lesions not observed during the training process, demonstrating its robustness and transferability to unseen damage types. ConclusionsDamage-aware biological language modeling combined with adaptive multimodal fusion enables accurate and interpretable identification of DNA lesions from nanopore sequencing data. This framework provides a scalable approach for characterizing genome-wide damage landscapes and illustrates how chemical DNA information can be systematically incorporated into genomic language models. The source code and pretrained models of this work are available at: https://github.com/UF-HOBIYin-Lab/DamageFormer.

genomics↗

bioRxiv2026

An extracellular vesicle biogenesis-inspired engineering platform for efficient protein delivery and therapeutic base editing

Efficient and controllable delivery of genome-editing proteins remains a central challenge for therapeutic translation of gene-editing technologies. Extracellular vesicles (EVs) offer an attractive non-viral delivery modality due to their biocompatibility and large capacity for cytosolic cargo delivery. Yet, rational strategies to achieve controlled and programmable protein loading are still lacking. Here, we present NEO-TOP-EVs, an EV biogenesis-guided engineering platform that systematically integrates key features of three design principles inspired by vesicle formation: 1) PI(4,5)P2-mediated plasma membrane targeting, 2) ESCRT-dependent membrane scission, and 3) self-assembly-driven cargo clustering for enabling efficient encapsulation of genome-editing ribonucleoproteins. Together, the NEO design increased cargo incorporation and enhanced functional delivery of gene editing modalities under particle-normalized conditions. Using NEO-TOP-EVs, we achieve efficient delivery of Cas9 and adenine base editor ribonucleoproteins without nucleic acid templates. In an in vitro proof-of-concept, delivery of an adenine base editor targeting proprotein convertase subtilisin/kexin type 9 (PCSK9) induces efficient splice-site disruption, resulting in reduced PCSK9 expression and enhanced LDL receptor activity. Proof-of-concept in vivo experiments provide preliminary evidence of functional Cre protein delivery to the liver. Together, these findings establish NEO-TOP-EVs as a modular platform for protein-based genome editing, demonstrating how biogenesis-informed EV engineering yields functional protein delivery at levels relevant to therapeutic development.

bioengineering↗

bioRxiv2026

Membrane-anchored influenza neuraminidase vaccine drives human-like broadly protective B cell responses

Influenza neuraminidase (NA) is a promising target for universal flu vaccines, yet eliciting potent B-cell responses against its conserved epitopes remains challenging. Here, we developed a membrane-anchored, folding-domain-free NA (mNA) that elicited superior head-specific germinal center B cell and antibody responses compared to soluble tetrameric NA. In non-human primates, mNA immunization induced cross-reactive memory B cell (MBC) responses, expanding clones with the conserved "DR" motif in HCDR3, a hallmark of human broadly reactive NA antibodies. These MBCs conferred cross-inhibitory activity against diverse NA variants and in vivo cross-protection. Cryo-EM analysis revealed that the 554-C2 clone targets the conserved enzymatic pocket via the "DR" motif, while the 554-C1 clone recognizes previously uncharacterized epitopes at the interface between two adjacent N2 monomers, effectively reducing plaque formation by contemporary H3N2 strains. Our findings highlight the immunological advantages of membrane-anchoring, providing a robust strategy for designing next-generation vaccines against influenza and other pathogens.

immunology↗