Mpox virus (MPXV) poses an increasing global health threat, as underscored by two World Health Organization declarations of Public Health Emergencies of International Concern, particularly after the emergence of a novel Clade Ib strain that exhibited high human-to-human transmissibility in the Democratic Republic of the Congo. However, the treatment options for MPXV infection remain extremely limited. To address this unmet need, we established an integrated platform combining single-cell transcriptomics and deep learning-based structural prediction to discover effective human monoclonal antibodies against MPXV. By integrating computational prediction with experimental validation, we identified five neutralizing antibodies targeting the following distinct viral forms: BA345, MA42, and MA49, which engage the extracellular enveloped virus-associated A35R glycoprotein; BAL31, which binds intracellular mature virus (IMV) protein A29L; and HB05, which targets IMV antigen H3L. Importantly, the elite monoclonal antibody BA345 conferred effective protection against MPXV and vaccinia virus both in vitro and in vivo. Combined in silico structure prediction and X-ray crystallography revealed a highly conserved epitope shared across orthopoxviruses. Surface plasmon resonance measurements revealed nanomolar equilibrium dissociation constants of BA345 for A35R homologs, corroborating its cross-reactive, broad-spectrum neutralizing activity against orthopoxviruses. Moreover, the BA345/BAL31 and MA49/BAL31 antibody cocktails developed in this study conferred robust therapeutic protection in MPXV-infected animals, substantially reducing disease severity and viral load. Our findings not only establish a practical paradigm for antibody discovery through the integration of deep learning-driven structure prediction with single-cell multiomics but also inform next-generation biodefense countermeasures against MPXV and related orthopoxviruses.
Asymmetric transcription of noncoding RNA LincGET is currently recognized as the earliest event regulating the first cell fate decision in mammalian embryogenesis. However, whether key protein factors modulate this process remains elusive. Here, we identify RBBP7 as the earliest protein factor regulating developmental cell fate in mammals. Loss of RBBP7 drives cells towards ICM lineage. In mouse late 2-cell embryos, unequal translation of Rbbp7 contributes to its asymmetric protein distribution, which subsequently induces inversed asymmetric histone acetylation H3K9ac by interaction with HDAC1, thereby promoting cell differentiation. Interestingly, RBBP7 and LincGET exhibit a consistent asymmetric tendency but direct different cell fates; depletion or overexpression of both Rbbp7 and LincGET restored the cell fate bias, suggesting a coordinated regulatory mechanism during initial lineage specification. In summary, our study reveals RBBP7 as a new protein factor and elucidates its role in the first cell fate decision.
Obesity and its associated metabolic complications represent a global health crisis, yet effective microbiota-targeted pharmacotherapies remain limited. Here, we report that GV-971 (sodium oligomannate), a marine-derived oligosaccharide originally developed for Alzheimer's disease, exerts potent anti-obesity and metabolic benefits by reprogramming gut microbial and host signaling networks. In high-fat diet-induced obese mice, GV-971 reduced adiposity, improved glucose homeostasis, and alleviated hepatic steatosis without affecting food intake. Multi-omics and causal intervention experiments revealed that GV-971 selectively decreased the abundance of Clostridium scindens, a keystone bacterium responsible for secondary bile acid synthesis. This decrease downregulated the expression of the baiF gene encoding 7α-hydroxysteroid dehydrogenase, leading to reduced intestinal deoxycholic acid (DCA) levels and inhibition of intestinal farnesoid X receptor (FXR) signaling. Restoration of C. scindens abundance, baiF expression, or DCA supplementation abrogated the metabolic benefits of GV-971, confirming the causal role of the C. scindens-DCA-FXR axis. Mechanistically, inhibition of intestinal FXR promoted thermogenic gene expression and white adipose tissue browning, thus enhancing systemic energy expenditure. These findings uncover a bacterium-metabolite-host signaling pathway underlying the effects of GV-971 and establish microbiota-directed FXR modulation as a promising therapeutic approach for obesity and metabolic disease.
Crimean-Congo hemorrhagic fever virus (CCHFV), designated by the WHO as a priority pathogen under its R&D Blueprint for emerging epidemics, poses a major global health threat, yet licensed vaccines or specific antiviral treatments are lacking. As the sole viral enzyme responsible for genome replication and transcription, the CCHFV L protein is a large, multienzymatic protein, but its exceptional size (> 450 kDa) and extensive domain architecture have hindered structural analysis. Here, we present high-resolution cryo-electron microscopy structures of the full-length CCHFV L protein in its apo state and bound to the 5' viral RNA promoter. These structures reveal the largest polymerase known among Bunyavirales and demonstrate that the apo form adopts a highly flexible conformation in which multiple functional elements remain disordered. Binding of the 5' promoter RNA triggers extensive conformational rearrangements that organize these elements into a catalytically competent active site. We define a conserved 5' hook-binding mode and identify two previously unrecognized residues (K1545 and E1637) that form a constriction at the NTP entry channel, representing newly defined regulatory motifs J and K conserved across Bunyavirales. We further characterize an expanded pendant domain unique to nairoviruses that, although not essential for promoter binding, likely modulates template movement within the internal tunnel during RNA synthesis. Our results provide the first structural framework for a nairovirus polymerase, illuminate the mechanisms of CCHFV RNA synthesis, and establish a foundation for structure-guided antiviral development against this high-priority pathogen.
Type 1 diabetes mellitus characterized by insulin deficiency and hyperglycemia is associated with female subfertility. However, how hyperglycemia affects the hypothalamic-pituitary-ovarian-uterine axis remains poorly understood. In this study, we performed single-cell transcriptomic profiling of the hypothalamus, pituitary, ovary and uterus during the proliferative phase of the menstrual cycle in type 1 diabetic macaques to systematically characterize changes in tissue-specific cellular heterogeneity, gene expression, and intercellular communication networks under diabetic conditions. Our analysis revealed significant upregulation of the TNF signaling pathway across multiple tissues, concomitant with marked activation of inflammation-related pathways. Notably, the macrophage migration inhibitory factor signaling pathway exhibited a tissue-specific regulatory pattern, being significantly upregulated in the hypothalamus and pituitary but downregulated in the ovary and uterus, suggesting divergent inflammatory modulation along the reproductive endocrine axis in response to diabetes. Moreover, we observed that diabetes leads to reduced FSHR expression during granulosa cell differentiation, and this process is further exacerbated by the upregulated expression of SFRP4, a known antagonist of follicle-stimulating hormone signaling, resulting in diminished granulosa cell responsiveness to follicle-stimulating hormone. Consequently, this dysregulation is correlated with increased FSHB expression in pituitary gonadotropes, likely due to disrupted ovarian feedback signaling. Collectively, our findings provide a comprehensive landscape of cellular and molecular alterations in immune and endocrine compartments in the female reproductive system in diabetic states, advancing our understanding of immune‒endocrine cell crosstalk in the context of metabolic disease.
Elevated intratumoral immune inflammation prior to treatment is typically associated with better outcomes in hot tumors treated with immune checkpoint inhibitors (ICIs). However, we observed a paradox in pMMR/MSS locally advanced rectal cancer (LARC) patients, where a subset with elevated baseline immune inflammation exhibited worse outcomes after combined radiotherapy and ICI treatment compared with patients with minimal immune inflammation. To investigate this counterintuitive finding, we performed paired scRNA-seq and scTCR-seq on longitudinally collected samples, including tumor biopsies (pre-treatment, post-radiotherapy, and post-immunotherapy) and peripheral blood mononuclear cells (pre-treatment and post-immunotherapy), from 20 pMMR/MSS LARC patients treated with sequential radiotherapy and ICI therapy (NCT06493240). We propose the concept of clonal entrapment to explain this phenomenon. Specifically, our profiling results reveal that increased HLA-DQA2 expression in dendritic cells and upregulated GDF15 expression in treatment-resistant tumor cells correlate with the restricted expansion of novel tumor-reactive TCR clonotypes. Consequently, the immune response is limited primarily by pre-existing TCR clonotypes within the tumor, especially those partially expanded under chronic inflammation, leading to the expansion of TCR clonotypes derived mainly from pre-treatment CD8+ T cell pools following ICI therapy. By identifying this feature of the pMMR/MSS LARC microenvironment, our study provides a high-resolution framework for understanding resistance to sequential radiotherapy and ICI therapy.
Abstract The recruitment and condensation of apoptosis-associated speck-like protein containing a CARD (ASC) are critical for ASC speck formation and inflammasome activation. However, how this process occurs efficiently in vivo remains unclear. Here, we identified the RNA helicase DDX6 as an ASC-interacting protein through immunoprecipitation‒mass spectrometry (IP‒MS) analysis. DDX6 promotes the activation of both NLRP3 and AIM2 inflammasomes by facilitating the recruitment of ASC to these receptors through its RNA helicase activity. Mechanistically, DDX6 functions as a scaffold protein for processing body (P-body) assembly and drives ASC speck formation in P-bodies via liquid‒liquid phase separation (LLPS). We report that membrane integrity is associated with stress granule (SG) formation and that in Caspase-1 –/– , Gsdmd –/– , or NINJ1-inhibited cells, DDX6 mediates initial ASC speck assembly in P-bodies, followed by their transition to SGs during inflammasome activation. DDX6 deficiency in macrophages increases host susceptibility to Listeria infection. Our results establish that DDX6 orchestrates ASC recruitment, speck formation, and subsequent transition through LLPS-mediated mechanisms, offering new insights into inflammasome assembly and potential therapeutic approaches for inflammasome-related diseases.
Manganese (Mn) has lingered in the shadows as a mere enzymatic cofactor, with its profound role in regulating the most fundamental life processes largely overlooked. This review heralds a "manganese renaissance" - a paradigm shift that elevates Mn from a passive trace element to a dynamic architect of metabolic homeostasis and a critical driver of disease. We synthesize breakthroughs that redefine its biological significance. In addition to enabling reactions for enzymes such as MnSOD, Mn actively governs lipid trafficking via the modulation of the COPII complex, facilitates cGAS/STING signaling for host immune responses, and precisely activates ion transporters and sensors to maintain cellular homeostasis. Dysregulated Mn homeostasis - whether stemming from genetic defects in key transporters (SLC30A10, SLC39A8, SLC39A11, and SLC39A14) or environmentally induced overload - fuels a spectrum of pathologies, including metabolic syndrome, Parkinsonism-like neurodegeneration, hepatic dysfunction, cardiovascular disease, and immune dysfunction. This disruption underscores the irreplaceable role of Mn as a biological linchpin, as its balance is not merely supportive but also central to sustaining health. In the future, we outline translational frontiers - from dietary Mn modulation and transporter-specific therapies for genetic Mn disorders to the elucidation of Mn signaling and the development of exposure guidelines to safeguard public health. This synthesis reaffirms that Mn is far more important than simply functioning as a nutrient. Research into Mn functions has been conducted across biology, environmental science, and medicine, and Mn acts as a master regulator whose emerging mechanisms will reshape our understanding of metabolic health and disease pathogenesis.
Compromised cortical inhibition during threat processing contributes to individual vulnerability to stress-related psychiatric disorders. However, the precise underlying neurobiological circuits remain elusive. Here, by combining monosynaptic viral tracing, electrophysiology, in vivo calcium imaging, and functional manipulations in mice, we elucidated a functionally specialized monosynaptic pathway originating from glutamatergic pyramidal neurons in the ventromedial prefrontal cortex (vmPFCGlu) to corticotropin-releasing hormone (CRH)-expressing neurons in the paraventricular nucleus of the hypothalamus (PVNCRH). We found that activating medial prefrontal cortex (mPFC)-driven CRH "pacemaker" cells propagated calcium signals within the local CRH network of the PVN. Hyperactivity of this vmPFCGlu-PVNCRH circuit promoted persistent social avoidance, consolidated threat memory, facilitated auditory-cued fear acquisition, and impaired extinction. Conversely, inhibition of this circuit selectively reduced social stress‑induced avoidance. Our findings define a novel top-down circuit that specifically controls psychosocial stress responses and amplifies susceptibility to conditioned fear.
The genetic architecture of glycemic dynamic metrics derived from continuous-glucose monitoring (CGM) across different populations remains poorly understood. Here, we conducted a trans-ethnic genome-wide association study (GWAS) meta-analysis of 20 CGM-derived glycemic traits, building upon a previously established European-ancestry CGM dataset and extending it through the inclusion of additional cohorts, in up to 9677 individuals originating from 2051 Chinese, 901 Dutch, and 6725 Israelis. Across 20 glycemic traits, we identified 18 genome-wide significant associations, of which 9 met study-wide significance, and three variants were novel. These variants indicated a shared genetic basis for continuous glycemic regulation and exhibited consistent patterns with those of sequential fingerstick glucose tests. Our findings further demonstrated that the identified genetic variants were enriched in pathways related to the nervous system. These findings were further supported by observed associations with brain magnetic resonance imaging (MRI) metrics, high CGM-related gene expression and co-regulation of quantitative trait loci in brain tissues. Additionally, we observed a positive relationship between genetic liability for the coefficient of variation (CV) and total cholesterol and a bi-directional putative causal relationship between hyperglycemia and type 1 diabetes across trans-ethnic populations. Moreover, we established a polygenic risk score (PRS) for additional participants and reported that certain glycemic traits were significantly associated with the risk of diabetes or pre-diabetes. These variants constituting the PRS demonstrated high transferability across general populations and pregnant women. Overall, our study yields unique insights into the high trans-ethnic and generalizable genetic architecture of CGM-derived glycemic profiles, supporting improved characterization of interindividual differences in glycemic dynamics and underscoring the potential for more personalized glucose management.
The ion permeation pathway is a critical determinant of ion channel function and selectivity; however, the structural basis for ion permeation in the P2X1 receptor, an ATP-gated ion channel crucial for platelet activation, thrombosis, and male infertility, remains incompletely understood. Here, we present high-resolution cryo-electron microscopy (cryo-EM) structures of the P2X1 receptor, which reveal a central ion permeation pathway spanning the entire extracellular domain, complementing the existing paradigms of ion channel architecture for the P2X receptor family. Within this pathway, we identify specific sites that coordinate hydrated calcium ions, including an aspartate ring that acts as a selectivity filter at the apex of the central vestibule. We also discover that a small molecule, 3,5-bis(trifluoromethyl)aniline, binds at the top of the central vestibule and potently inhibits cation flux through this central permeation pathway. Our findings reveal a new inhibitor-binding site in the P2X1 receptor. These insights provide a structural framework for the rational design of subtype-specific P2X receptor inhibitors targeting the central vestibule.
The activation of brown adipose tissue (BAT) for thermogenesis represents a crucial physiological mechanism that helps maintain body temperature during cold exposure. Nevertheless, the exact mechanisms underlying the sustained activation of BAT under cold conditions remain incompletely understood. In this study, we reveal that soluble ST2 (sST2) mediates a white adipose tissue (WAT)-to-BAT endocrine mechanism that is essential for the continuous activation of BAT during cold exposure. Specific depletion of sST2 blocks alternative thermogenesis following BAT denervation and renders mice sensitive to cold during prolonged cold exposure. Mechanistically, sST2 is induced and secreted from epididymal white adipose tissue (eWAT) upon the activation of Creb1, which is driven by β1 and β2 adrenergic receptor signaling. Secreted sST2 directly binds to the β3 adrenergic receptor in BAT and, in synergy with norepinephrine, induces BAT thermogenesis independent of IL33. Additionally, supplementation with sST2 promotes beige fat formation. Therefore, our study illustrates a novel mechanism through which the adipokine sST2 derived from eWAT mediates sustained BAT activation during cold exposure through the integration of neural and humoral signals. More importantly, sST2 exerts a synergistic effect on BAT activation when combined with β3-adrenergic receptor agonists.
