Biology subjects

Wang, Q.

Publications and source records attributed to Wang, Q..

At least 55 records · Page 3Linked to original sources

bioRxiv2026

Therapeutic targeting of MYC- and MYCN-driven medulloblastoma with a novel MYC degrader molecule

BackgroundMedulloblastoma (MB) is the most common malignant brain tumour in children, and aggressive subgroups are frequently driven by the oncoproteins MYC or MYCN. Direct therapeutic targeting of MYC/MYCN has been challenging because of their intrinsically disordered protein structures. The aim of this study was to determine whether novel SE486-11 analogues (UNSW-SCs) can therapeutically target MYC/MYCN-driven MB. MethodsThe anticancer activity of UNSW-SCs was assessed in MB cell lines with differential MYC/MYCN expression. Target engagement was evaluated using surface plasmon resonance and drug affinity responsive target stability assays. Blood-brain barrier penetration, MYC/MYCN protein degradation, cell cycle effects, apoptosis, DNA damage, and synergy with histone deacetylase (HDAC) inhibitors were examined. Therapeutic efficacy was evaluated in murine models of MYC- and MYCN-driven human MB. ResultsUNSW-SCs showed potent anticancer activity, with preferential selectivity toward MB cells expressing high MYC/MYCN levels and IC50 values ranging from 0.22 to 1.18 M. The lead molecule, UNSW-SC-22, directly bound MYC, crossed the blood-brain barrier, and achieved a brain-to-plasma ratio of 1.44 at peak concentrations. UNSW-SC-22 induced MYC/MYCN-dependent cytotoxicity associated with enhanced proteasomal degradation, cell cycle arrest, apoptosis, and DNA damage. Combined treatment with HDAC inhibitors further reduced MYC/MYCN protein levels, increased DNA damage, and enhanced apoptosis. In vivo, UNSW-SC-22, either alone or with entinostat, significantly suppressed intracranial tumour growth and prolonged survival. ConclusionsUNSW-SC-22 is a brain-penetrant MYC/MYCN-targeting molecule with potent preclinical activity in MYC/MYCN-driven MB, supporting its development as a monotherapy or combination strategy with HDAC inhibition. Key PointsO_LIUNSW-SC-22 directly targets MYC/MYCN in medulloblastoma. C_LIO_LIUNSW-SC-22 crosses the blood-brain barrier and prolongs survival. C_LIO_LIHDAC inhibition enhances UNSW-SC-22 activity in MYC-driven cells. C_LI Importance of the StudyMYC- and MYCN-driven medulloblastomas remain among the most aggressive paediatric brain tumours, yet direct pharmacological targeting of MYC/MYCN has historically been difficult. This study identifies UNSW-SC-22 as a novel brain-penetrant small molecule that directly engages MYC/MYCN and promotes proteasomal degradation, leading to tumour cell death. Compared with prior MYC-directed approaches, UNSW-SC-22 combines direct target engagement, blood-brain barrier penetration, and in vivo efficacy in orthotopic MYC- and MYCN-driven medulloblastoma models. The study further demonstrates that combining UNSW-SC-22 with histone deacetylase inhibition enhances MYC/MYCN suppression, DNA damage, apoptosis, and survival benefit. These findings provide a translational framework for developing MYC/MYCN degradation as a therapeutic strategy for high-risk medulloblastoma and support future pharmacological optimisation, biomarker-guided patient selection, and combination therapy development.

cancer biology↗

bioRxiv2026

Kidney medulla macrophages maintain a free flow of urine by sensing force

Blockade by sedimentary particles, such as mineral crystals, is a continuous risk the kidney tubule faces. To prevent that, kidney resident macrophages form transepithelial protrusions and remove intratubular sedimentary particles, a behavior particularly prevailing in the medulla over the cortex. However, the molecular mechanisms underlying this characteristic behavior of medulla macrophages are incompletely understood. In this study, we identified that the medulla had higher mechanical stiffness than the cortex in steady state, which was further elevated when kidney stone formed. Increased tissue rigidity was sensed by medulla macrophages via mechanoreceptor Piezo1, which promoted macrophage protrusion formation and their ability to clean the tubules. Loss of Piezo1 expression in kidney macrophages predisposed mice to intratubular accumulation of mineral crystal in steady state and accelerated kidney stone formation during oxalate intake challenge. Signaling via Piezo1 mobilized molecules involved in cell adhesion and protrusion assembly, including Talin2 and focal adhesion kinase (FAK). Finally, we developed a first-of-its-kind cell-based therapy for the treatment of experimental nephrolithiasis by exploiting macrophage Piezo1 activity, and this strategy shows great promise for future translational research.

physiology↗

bioRxiv2026

Internalized Components of Membrane Attack Complexes Disrupt Proteostasis and Acquire Alarmin-Like Properties

Immune effects of membrane attack complexes (MAC) have been widely attributed to their abilities to cause cell death. Here, we show that the MAC component, C9, forms non-cytolytic aggregates with pro-inflammatory effects. Intracellular aggregates of C9 are detected within inflamed tissues of patients in association with endothelial cell (EC) activation but not increased cell death. We identify NUMBL as a Rab35 effector that directly binds surface-bound C9 to promote C9 internalization and entry into the endolysosomal pathway. Within acidified endolysosomes, C9 forms insoluble aggregates that are targeted for degradative aggrephagy in a process that activates NF-{kappa}B. For C9 aggrephagy to occur, ZFYVE21, a Rab5 effector, complexes with RNF34 to bridge C9 aggregates to LC3B+ aggresome membranes. We detect C9 aggregates in vivo, and we show that a ZFYVE21-RNF34 signaling axis is required for C9 aggrephagy and NF-{kappa}B -dependent EC activation in three separate mouse models. Mice with conditional loss of ZFYVE21 in ECs show reduced aggregraphy, resulting in attenuated systemic inflammation and reduced tissue injury following skin transplantation. Our data show that the C9 component of MACs forms intracellular aggregates with alarmin-like properties.

biochemistry↗

bioRxiv2026

Reverse genetics in humanized mice reveals CARD8-mediated pyroptosis causing pancytopenia in human DPP9 deficiency

Loss of function mutation in the human DPP9 gene causes Hatipoglu syndrome leading to severe inflammasomopathy. A key feature of the disease is pancytopenia and patients require bone marrow transplantation, but the mechanism of cell loss is unclear since Dpp9 mutant mice have normal hematopoiesis, suggesting that a distinct mechanism of disease occurs in humans. Here, we present a model of human DPP9 deficiency leveraging reverse genetics in the MISTRG6 humanized mice. We found that CRISPR editing of human CD34+ hematopoietic stem and progenitor cells (HSPCs) led to very efficient and persistent gene deletion in vivo. Human DPP9 deletion recapitulated cytopenia in peripheral blood and in the bone marrow, and cell loss was cell intrinsic. However, DPP9 deletion led to little transcriptional changes suggesting post-transcriptional regulation in human HSPCs. Mechanistically, DPP9 deficiency led to the activation of the CARD8 inflammasome resulting in HSPC pyroptosis, whereas NLRP1 was dispensable for cell death. Thus, our results reveal a unique human mechanism of disease and offer therapeutic insight for this inflammasomopathy.

immunology↗

bioRxiv2026

The kinetochore proteins Ndc80 and Dsn1 are required for survival of postmitotic neurons

Dsn1 and Ndc80 are essential proteins of the kinetochore complex and required for chromosome segregation in dividing cells and for regulating development and microtubule dynamics in postmitotic neurons. With conditional deletion of floxed alleles, we here show that Dsn1 and Ndc80 are also required for the viability of postmitotic neurons, both in cultures of hippocampal and cortical neurons and in vivo in the retina. Loss of these proteins triggers apoptosis, as indicated by caspase cleavage and an increase in nuclear DNA breakage. The pro-survival function of the kinetochore components is distinct from that which was previously demonstrated for regulation of neuronal synaptogenesis. The microtubule-binding domain of Ndc80 is required for the synaptogenic functions but Ndc80 lacking this domain can nonetheless rescue the viability of neurons from which Ndc80 has been deleted. Similarly, whereas the synaptogenic function involves regulation of microtubules in axons and dendrites, a nucleus-localized Dsn1 is sufficient to rescue the viability of neurons from which Dsn1 has been deleted. Thus, postmitotic neurons retain a nuclear requirement for components of the kinetochore in order to prevent apoptosis.

neuroscience↗

bioRxiv2026

PKD2 structural destabilization drives primary cilia degeneration and ADPKD pathogenicity.

Abstract/SummaryHuman variants in renal polycystins (PKD1, PKD2) are responsible for most forms of autosomal dominant polycystic kidney disease (ADPKD), a common genetic disorder without curative drug treatment. Renal polycystins form ion channels in primary cilia, but our understanding of their molecular dysregulation caused by disease-associated variants is limited. Using cryo-electron microscopy (cryo-EM), primary cilia electrophysiology and super-resolution analysis, we investigated the mechanistic impact and pathogenic potential of the disease-associated PKD2 missense variant (D511V) located within the channels voltage sensor domain (VSD). Our findings define how this mutation neutralizes critical transmembrane charge interactions, which attenuates PKD2 protein stability resulting in abolished ciliary channel trafficking and function in membranes. To assess the pathogenic effect of this variant in vivo, we generated novel mouse strains carrying the analogous PKD2 mutation in combination with a conditional floxed allele (Pkd2D509V/fl) that exhibit renal tubule primary cilia degeneration and develop rapid renal cysts. Our results establish a clear direct correlation between the in vitro molecular dysfunction and phenotypic in vivo consequences while providing a valuable tool to evaluate ADPKD therapeutic interventions. Translational StatementADPKD is a genetic kidney disorder affecting millions of patients globally and is primarily caused by variants in renal polycystin genes (PKD1, PKD2). Polycystins function as ion channel subunits in primary cilia but the mechanistic impact and cystogenic propensity of disease-associated variants remain poorly defined. The authors employ advanced methodologies including cryo-EM to uncover distinct structurally destabilizing effects of a human PKD2 mutation, while generating a new mouse model which genetically expresses the same variant and recapitulates the human disease. The findings define primary cilia degeneration results from PKD2 hypostasis and establish new tools to assess ADPKD therapies. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=107 SRC="FIGDIR/small/734313v1_ufig1.gif" ALT="Figure 1"> View larger version (50K): org.highwire.dtl.DTLVardef@1cd02ceorg.highwire.dtl.DTLVardef@73410eorg.highwire.dtl.DTLVardef@b40691org.highwire.dtl.DTLVardef@11b5a93_HPS_FORMAT_FIGEXP M_FIG C_FIG

physiology↗

bioRxiv2026

Hippocampal-cortical coupling dynamics drive system consolidation of remote memory

System consolidation transforms temporary hippocampal representation of memory into long-term storage in cortex. The underlying neural substrate, however, remains enigmatic. Here, we tracked the spatiotemporal evolution of hippocampus (HPC)-cortex local field potentials and single-neuron spikes in behaving animals during fear memory formation. During learning, HPC fast gamma exhibited a progressive phase shift relative to PFC theta oscillations, with gamma power aligning to progressively later phases of the PFC theta cycle. Strikingly, a related phase-shifted coupling pattern re-emerged during subsequent consolidation in association with hippocampal sharp-wave ripples and transient PFC spindle events during NREM sleep. Across this process, interregional interactions evolved from HPC-driven cortical gamma coherence at recent stages to PFC-mediated cortical low-frequency coherence at remote stages. Using closed-loop optogenetic perturbations, we demonstrated a stepwise causal chain of coupling events underlying remote memory formation. Our study revealed HPC-PFC coupling phase shift as a feasible substrate mediating recent-to-remote transformation of memory.

neuroscience↗

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

Benchmarking cell type annotation in spatial transcriptomics: resolving cellular hierarchies, biological fidelity, and dynamic cell states

Spatial transcriptomics enables the quantification of gene expression within its native tissue context, providing unprecedented insight into tissue architecture, cellular ecosystems, and local cell-cell interactions at regional and single-cell resolution. Accurate cell type annotation is a critical prerequisite for interpreting these data and is often the first and most essential step in downstream analysis. Despite rapid advances in computational methods, cell type annotation remains challenging and frequently requires extensive expert-driven manual curation based on marker-gene expression, spatial context, and prior biological knowledge. While early approaches relied primarily on transcriptional similarity, newer methods increasingly incorporate spatial information, histological features, and multimodal data to improve annotation accuracy. Nevertheless, reliable annotation remains difficult when biological interpretation requires fine-grained subtype resolution, particularly for platforms with limited gene panels, tissues undergoing dynamic cellular state transitions, and studies in which reference and query datasets differ substantially in biological context or technical modality. Here, we present a systematic benchmark of 20 state-of-the-art cell type annotation methods across four spatial transcriptomics datasets spanning diverse technologies, experimental conditions, cell numbers, and gene panel sizes. Importantly, all benchmark datasets contain expert-curated cell type labels, including wellresolved cell populations and subtype annotations, providing high-quality biological ground truth for evaluation. The benchmark encompasses both reference-based and reference-free methods representing a broad range of computational frameworks. Performance was assessed using conventional classification metrics, including accuracy and F1-based measures, together with structure-aware metrics that evaluate both cell-level annotation accuracy and preservation of higher-order biological organization. Across datasets, annotation performance varied substantially according to tissue context, reference-query similarity, and annotation granularity. Fine-grained subtype annotation and recovery of rare cell populations remained challenging for many methods, particularly in datasets capturing injury, repair, developmental, and regenerative processes characterized by continuous cellular state transitions. Notably, high classification accuracy did not necessarily correspond to preservation of global cellular relationships or biologically coherent downstream pathway and gene-set enrichment analyses. Overall, scANVI, Seurat, and TACCO consistently ranked among the top-performing methods, although their relative advantages were context dependent. Together, our results provide a comprehensive assessment of current annotation strategies for spatial transcriptomics and offer practical guidance for selecting methods that best align with specific biological questions, dataset characteristics, and analytical priorities.

bioinformatics↗

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

Targeting Lysosomal MCOLN1/TRPML1 Ion Channels to Finely Alleviate Diabetes Mellitus via a Ca 2+ - CaMKKβ-AMPK pathway

Type 2 Diabetes mellitus (T2DM) is a metabolic syndrome characterized by hyperglycemia and various complications. Current drugs are limited by side effects and resistance, necessitating novel targets and therapies. Previous studies have shown that MK-83, a synthetic agonist of transient receptor potential mucolipin 1 (TRPML1/ MCOLN1), a lysosomal Ca2+ channel, activates adenosine 5'-monophosphate-activated protein kinase (AMPK), a key therapeutic target in diabetes. However, whether targeting TRPML1 can treat T2DM remains unclear. In this study, we found that transgenic overexpression or pharmacological activation of TRPML1 finely controls AMPK phosphorylation via a Ca2+-CaMKK{beta}-dependent mechanism. This activation promotes glucose transporter 4 (GLUT4) translocation and dramatically increases intracellular glucose uptake. Conversely, genetic inactivation or pharmacologically inhibition of TRPML1 blocks both AMPK activation and glucose uptake. More importantly, pharmacological activation of TRPML1 in vivo dramatically alleviates hyperglycemia in db/db mice (a T2DM model), as evidenced by random blood glucose levels, fasting blood glucose levels and HbA1c levels. Furthermore, other hallmark features of db/db mice-including impaired oral glucose tolerance, reduced insulin tolerance, and elevated ALT and AST levels- were all ameliorated upon TRPML1 activation. Hence, targeting lysosomal TRPML1 channel represents a promising therapeutic strategy for T2DM, with highly specific TRPML1 agonists as potential anti-diabetic agents.

Cell Biology↗

bioRxiv2026

Dehydration/1,6-addition-based Site-specific Bioconjugation Unveils Norepinephrinylation as a Widespread Post-translational Modification in the Cellular Proteome

Norepinephrine (NE) is a key neurotransmitter and hormone involved in diverse physiological and pathological processes. Beyond its canonical non-covalent signaling through binding adrenergic receptors, NE also induces protein post-translational modifications (PTMs), representing an emerging regulatory mechanism. Two major forms of NE-derived PTMs have been proposed: non-enzymatic norepinephrinylation (NEylation) of cysteine residues mediated by NE quinone and transglutaminase 2 (TG2)-catalyzed NEylation of glutamine residues. However, the cellular abundance, biochemical basis, and pathophysiological roles of NEylation remain poorly understood due to limited detection tools. Here, we report a novel bioconjugation chemistry for selective labeling and enrichment of the endogenous NEylation proteome in cell lines and tissues, which is based on acid-catalyzed dehydration and 1,6-addition to thiol probes. This strategy enables fluorescence imaging and chemical proteomic profiling, revealing NEylation as a widespread PTM that affects enzymatic activities of modified proteins, including protein tyrosine-protein phosphatase non-receptor type 11 (PTPN11). Visual Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=75 SRC="FIGDIR/small/731782v3_ufig1.gif" ALT="Figure 1"> View larger version (21K): org.highwire.dtl.DTLVardef@406ac5org.highwire.dtl.DTLVardef@121d6b9org.highwire.dtl.DTLVardef@98354corg.highwire.dtl.DTLVardef@d312ef_HPS_FORMAT_FIGEXP M_FIG C_FIG

biochemistry↗

bioRxiv2026

Structural basis for loop-dependent inhibition by AcrIIC1 anti-CRISPR proteins

Most known anti-CRISPR proteins exert inhibition via solitary functional pathways, and few natural Acr homologs have been reported to possess coupled inhibitory mechanisms. Here, we demonstrate that AcrIIC1Boe represses Nme1Cas9 activity through loop-coupled inhibitory effects. Beyond canonical occlusion of the HNH catalytic site, the unique extended loop of AcrIIC1Boe further perturbs sgRNA binding and restricts R-loop maturation to reinforce inhibition. Strikingly, loop deletion completely abrogated this layered inhibitory capacity, confirming that the loop is indispensable for full anti-CRISPR function. Consistently, transplantation of this functional loop into the weakly active AcrIIC1Nme1 markedly upgraded its inhibitory performance. Our findings reveal an unprecedented loop-governed coupled inhibition mode, clarify the structural basis for functional divergence within the AcrIIC1 family, and provide a modular engineering strategy for the rational design and optimization of high-potency anti-CRISPR proteins.

Molecular Biology↗

bioRxiv2026

Retinoid signaling promotes frontonasal identity while repressing maxillary identity during craniofacial development

Vertebrate facial development depends on the correct specification of embryonic facial prominences, a process known to be governed by region-specific reciprocal signaling pathways between the craniofacial ectoderm, endoderm and mesenchyme. This process is further modulated by transcriptional and epigenetic mechanisms that regulate the expression of essential genes in mesenchymal progenitors derived from cranial neural crest cells. Retinoid signaling plays a critical role in facial development, and both gain-and loss-of-function results in a wide spectrum of facial defects, including orofacial clefts. In this study, we identified retinoid signaling as a critical regulator of cranial neural crest cell specification toward a frontonasal process identity in mice. Rdh10 oxidizes vitamin A (all-trans retinol) to retinal, which is a rate limiting step in the synthesis of retinoic acid. Rdh10 loss-of-function resulted in ectopic formation of whisker pads - a derivative of the maxillary process - within the frontonasal process region. This transformation was evidenced by the mis-expression of maxillary specific transcription factors including Meis2 and Lhx6 in the frontonasal mesenchyme. Furthermore, these transcription factors exhibited increased chromatin accessibility at their consensus binding sites following the loss of retinoid signaling in frontonasal cranial neural crest cells. These results indicate that retinoid signaling acts as a critical regulator specifying frontonasal identity and fate of cranial neural crest cells as they migrate into the frontonasal process, while concomitantly repressing maxillary process fate. These results not only advance our understanding of frontonasal prominence specification and the evolutionary development of craniofacial structures but also offers valuable insights into the etiology and pathogenesis of craniofacial malformations such as orofacial clefts.

developmental biology↗

bioRxiv2026

Lack of ancestral SARS-CoV-2 imprinting promotes BA.3.2.2 infection in children

The ongoing evolution of SARS-CoV-2 is heavily constrained by population-level immune imprinting. Recent genomic surveillance reveals an unexpected demographic shift: the highly mutated BA.3.2.2 sublineage is significantly enriched in paediatric populations globally, unlike concurrent variants XFG and NB.1.8.1. Here, the evidence demonstrates that this paediatric susceptibility arises from the absence of ancestral Wuhan-strain immune imprinting. Serological analyses reveal a profound failure to neutralise BA.3.2.2 in young children with no ancestral strain exposure. Concurrently, single-B-cell sequencing analyses demonstrate that BA.3.2.2 completely evades Omicron-specific Class 1/4 neutralising antibodies, encoded by IGHV2-5 and IGHV5-51, which dominate the repertoires of weakly imprinted individuals such as young children. Conversely, BA.3.2.2 remains uniquely susceptible to broadly cross-reactive Class 1 antibodies, encoded by IGHV3-53/66, typically enriched in adults with strong ancestral imprinting, such as mRNA vaccine recipients. Critically, sustained transmission of BA.3.2.2 in paediatric populations may catalyse the emergence of secondary variants that combine the paediatric-evading features of BA.3.2.2 with adult-evading Class 1 mutations. This could allow the lineage to breach the imprinted immunity of the adult population, potentially driving widespread global transmission. Together, these findings show that legacy immune imprinting paradoxically provides superior protection against a highly divergent saltation variant, directly explaining the age-biased transmission of BA.3.2.2, and informing future paediatric vaccination strategies.

immunology↗

bioRxiv2026

Multi-Omics Characterization of Human Molecular Responses to Spaceflight Across Two Independent Missions

Human spaceflight has historically been led by government agencies, but the emergence of commercial organizations is enabling broader participation and new research opportunities. In this study, we present a comprehensive molecular characterization of early human responses to spaceflight, leveraging multi-omics data across the first three weeks of two commercial missions. Biospecimens from six individuals, four from the Axiom 2 mission (10 days) and two from Axiom 3 (21 days), were analyzed using single-cell and bulk RNA sequencing, alongside proteomic profiling. Individual and integrative analyses of these datasets reveal systemic changes in cell types, transcripts, and proteins related to immune regulation, osteoclast differentiation, NF-{kappa}B signaling, and blood homeostasis pathways. Importantly, several of the detected pathways align with physiological patterns observed in longer-duration missions. This work establishes a foundational resource for understanding early adaptation to spaceflight at the cellular and molecular levels, providing insights to reduce future space-travel health risks. HighlightsO_LIFirst integrated multi-omics analysis using single-cell, bulk RNA sequencing and proteomic profiling of early human spaceflight responses across two commercial missions (Ax-2 and Ax-3). C_LIO_LIDistinct PBMC clustering was observed across Ax-2 and Ax-3, and post-flight samples. It showed fewer monocytes, dendritic cells, and megakaryocytes with increased naive CD4 and cytotoxic T cells. C_LIO_LIMulti-omics integration identifies shared biological signatures, including osteoclast differentiation, metabolic stress, and coagulation changes. C_LIO_LIThese findings lay the foundation for developing countermeasures to protect immune, skeletal, and vascular health during spaceflight. C_LI

molecular biology↗

bioRxiv2026

Tracing the Evolutionary Origin of Interferon to Basal Chordates and Unveiling Its Antiviral Functionality in Jawless Vertebrates

Interferons (IFNs) are essential mediators of antiviral defense in vertebrates, having gradually replaced the RNA interference (RNAi) antiviral mechanism that predominates in invertebrates and plants. To date, IFNs have been identified exclusively in jawed vertebrates, leaving the origin of IFN-based antiviral mechanisms largely mysterious. In this study, by conducting a genome-wide screening of IFN homologs accross various species, we successfully identified seveal IFN homologs from agnatha and lancelet, but not other invertebrates. Notably, both agnatha and lancelet IFN homologs have ability to induce a set of interferon-stimulated genes (ISGs)-like genes, thus tracing the origin of IFN to basal chordate. Using VSV infected peripheral blood mononuclear cells (PBMCs) of Japanese lampreys, we found that lamprey IFNs have antiviral functionality by inducing the expression of hundreds of ISGs through interacting with a heterodimeric complex composed of CRFB7 and CRFB14. In addition to robustly mediating antiviral responses in monocytes, lamprey IFNs exert their effects on variable lymphocyte receptor B (VLRB)+ cells by remodeling the cytokine/chemokine networks to orchestrate antiviral innate and adaptive immunity. Furthermore, cross-species functional comparison of Dicer revealed that changes in residues essential for dsRNA processing occurred concurrently with the evolution of the IFN system. Collectively, these findings uncover the evolutionary origin of IFN and underscore its ancient roles in antiviral response and immune regulation, especially in the takeover of the RNAi antiviral mechanism during the early evolution of vertebrates. Significance StatementInterferons (IFNs) represent a hallmark of vertebrate antiviral immunity, yet their origin has remained elusive. Here, we reported bona fide IFN ligands and cognate receptors in both lamprey and lancelet, two key species placed at the transition from invertebrates to vertebrates. Using the VSV infection model, we further demonstrated the roles of lamprey IFN in antiviral defense and immune regulation. We also found that substitutions in residues essential for Dicer mediated dsRNA processing coincided with the emergence of the IFN system. Collectively, our findings provide key insights into the evolutionary origin of IFN-based immunity and its gradual replacement of RNAi as the dominant antiviral strategy in vertebrates.

evolutionary biology↗

bioRxiv2026

Enemies, more than sex, shape butterfly post-mating odor evolution

Not all odors influencing mating behavior evolve as sex pheromones. Female butterflies post-mating odors have been considered species-specific anti-aphrodisiac pheromones shaped by sexual selection but may also serve broader ecological roles shaped by natural selection. Males transfer odors to females that repel rivals, yet the widespread use of these compounds across phyla makes them targets for eavesdropping, such as by phoretic egg parasitoids. We show that in cabbage white butterflies (Pieris spp.), these odors are highly variable and attract parasitoids, deter predators, and influence oviposition. Using gas chromatography and electroantennography, we demonstrate that odor emission and perception lack species-specificity: compounds once thought unique to P. brassicae and P. rapae are shared across Pieridae. In P. napi, odor variation among populations correlates with parasitoid pressure, but not with latitude, genetic distance, or mating frequency, suggesting ecological rather than sexual drivers. In P. brassicae, CRISPR/Cas9 disruption of odor perception alters oviposition and increases susceptibility to parasitism. Moreover, these odors render females unpalatable to birds. Together, our results show that post-mating odors in Pieris butterflies may act as aposematic signals. We provide evidence that these signals evolve under multiple selective pressures, balancing deterrence of mates and predators, parasitoid avoidance, and host-plant interactions. These findings suggest that chemical signals should be viewed as integrating ecological and reproductive pressures, rather than being interpreted solely through the lens of sexual communication.

ecology↗