Biology subjects

He, J.

Publications and source records attributed to He, J..

At least 127 records · Page 7Linked to original sources

bioRxiv2025

m5C RNA Methylation Is Dysregulated by Oncogenic Herpesviruses via c-Myc Signaling to Counteract Host Antiviral Factors

Herpesviruses are a group of double-stranded DNA viruses known to develop versatile viral strategies to escape host immune surveillance for promoting their replication and propagation. This is illustrated by Kaposis sarcoma-associated herpesvirus (KSHV), an oncogenic gamma-herpesvirus that overcomes host immune suppression by multiple mechanisms. In this study, we reported that KSHV dysregulates 5-methylcytosine (m5C) modification and mRNA stability of host antiviral factors to benefit its lytic replication. KSHV lytic reactivation or de novo challenge led to downregulation of m5C RNA methyltransferases, NSUN2 and NSUN1 (NSUN2/1), while NSUN2/1 depletion promoted KSHV lytic replication. Such KSHV-mediated downregulation of NSUN2/1 is via suppression of the transcriptional factor c-Myc. We further performed the RNA bisulfite sequencing (RNA-BS-seq) to identify KSHV-dependent m5C modification of host mRNAs. KSHV lytic reactivation led to the significant reduction of m5C methylation and mRNA stability of TRIM25, a key activator of the RIG-I pathway, while TRIM25 depletion indeed promoted KSHV lytic replication. These host-virus interaction events were also observed in the infection of another oncogenic gamma-herpesvirus Epstein-Barr virus (EBV). Overall, our results highlighted a new strategy for human gamma-herpesviruses to counteract host antiviral factors and promote their lytic replication by manipulating host m5C RNA methylation. Significance StatementOur study has identified a novel viral mechanism of human gamma-herpesviruses to manipulate host RNA methylation machineries to subvert immune defenses and enhance viral lytic replication. In particular, our new data showed that KSHV/EBV downregulate the key 5-methylcytosine (m5C) RNA writers NSUN2/1 via c-Myc, and thus decrease m5C modification and stability of TRIM25 mRNA. As TRIM25 is a key E3 ubiquitin ligase in RIG-I signal transduction, its inhibition disrupts RIG-I mediated antiviral sensing and thus favors viral lytic replication. As human gamma-herpesviruses are critical pathogens that highly associate with multiple human diseases especially certain tumors, such studies are significant to shed light in improving the fundamental understanding of virus-host interactions and identifying new host targets for future translational applications.

microbiology↗

bioRxiv2025

Differential Regulation of Hepatic Macrophage Fate by Chi3l1 in MASLD

Metabolic dysfunction-associated steatotic liver disease (MASLD) progression involves the replacement of protective embryo-derived Kupffer cells (KCs) by inflammatory monocyte-derived macrophages (MoMFs), yet the regulatory mechanisms remain unclear. Here, we identify chitinase 3-like 1 (Chi3l1/YKL-40) as a critical metabolic regulator of hepatic macrophage fate. We observed high expression of Chi3l1 in both KCs and MoMFs during MASLD development. Genetic deletion of Chi3l1 specifically in KCs significantly exacerbated MASLD severity and metabolic dysfunction, whereas MoMF-specific Chi3l1 deletion showed minimal metabolic effects. Mechanistic studies revealed that this cell type-specific regulation arises from differential metabolic requirements: KCs display elevated glucose metabolism compared to MoMFs. Chi3l1 directly interacts with glucose to inhibit its cellular uptake, thereby selectively protecting glucose-dependent KCs from metabolic stress-induced cell death while having negligible effects on less glucose-dependent MoMFs. These findings uncover a novel Chi3l1-mediated metabolic checkpoint that preferentially maintains KCs populations through glucose metabolism modulation, providing important new insights into the pathogenesis of MASLD and potential therapeutic strategies targeting macrophage-specific metabolic pathways.

molecular biology↗

bioRxiv2025

cellSTAAR: Incorporating single-cell-sequencing-based functional data to boost power in rare variant association testing of non-coding regions

Whole genome sequencing (WGS) studies have identified hundreds of millions of rare variants (RVs) and have enabled RV association tests (RVATs) of these variants with complex traits and diseases. Analysis of non-coding variants is challenged by the considerable variability in regulatory function which candidate Cis-Regulatory Elements (cCREs) exhibit across cell types. We propose cellSTAAR, which integrates WGS data with single-cell ATAC-seq data to capture variability in chromatin accessibility across cell types via the construction of cell-type-specific functional annotations and variant sets. To reflect the uncertainty in cCRE-gene linking, cellSTAAR also links cCREs to their target genes using an omnibus framework which aggregates results from a variety of popular linking approaches. We applied cellSTAAR on Freeze 8 (N = 60,000) of the NHLBI Trans-Omics for Precision Medicine (TOPMed) consortium data to four lipids phenotypes: LDL cholesterol, a binary variable corresponding to high LDL cholesterol, HDL cholesterol, and triglycerides. We also provide replication results for all four phenotypes using UK Biobank (N = 190,000). Evidence from simulation studies and our real data analysis demonstrates that cellSTAAR boosts power and improves interpretation of RVATs of cCREs.

genetics↗

bioRxiv2025

Senescence-induced reparative fibroblasts enable scarless wound healing in aged murine skin.

It has been reported that elderly individuals exhibit reduced scarring during the wound healing process compared to younger adults. However, the underlying mechanisms responsible for this phenomenon remain poorly understood. Here, we revealed that aged mice exhibited more pronounced regenerative outcomes compared to young mice, characterized by increased hair follicle numbers and collagen fiber features closer to normal skin. Single-cell sequencing identified a reparative fibroblast subpopulation (Prss35+Fib) enriched in the aged group, which promotes regeneration through communication with epithelial cells, macrophages, and T cell subpopulations via PTN and EREG signaling. Spatial transcriptomics validated this communication pattern by elucidating cell proximity and locating the regenerative niche in the upper dermis. Finally, EREG treatment significantly enhanced regenerative outcomes in young mice, while the small wound model of aged mice, lacking the reparative fibroblast and EREG signaling, failed to achieve regeneration. Collectively, our findings advance the understanding of regenerative plasticity in aging and provide new insights for designing scarless therapeutic strategies.

cell biology↗

bioRxiv2025

The novel RNA polymerase I transcription inhibitor PMR-116 exploits a critical therapeutic vulnerability in a broad-spectrum of high MYC malignancies.

Ribosome biogenesis (RiBi) is a key determinant of cell growth and proliferation and is highly elevated in cancer due to the activation by oncogenes such as MYC. First-generation RiBi inhibitor CX-5461, while demonstrating clinical potential for cancer treatment, also induces DNA damage through off-target inhibition of TOP2 and potentially other mechanisms, bringing into question RiBi as a target for cancer therapy. In this study, we test second-generation RiBi inhibitor, PMR-116. PMR-116 exhibits improved drug-like properties compared to first-generation RiBi inhibitors and has robust anti-tumour activity in the absence of global DNA damage signalling in a broad range of pre-clinical models of haematologic and solid cancers, particularly in malignancies where MYC is either the driver of disease or is elevated. Thus, our work demonstrates that RiBi is a genuine target for cancer therapy and highlights the potential to exploit a critical therapeutic vulnerability in high-MYC human cancers with dismal therapeutic outcomes. Statement of significanceDespite the development of new cancer therapies, most advanced malignancies remain incurable. We demonstrate that PMR-116, a second-generation RiBi inhibitor, has robust therapeutic efficacy in preclinical models of cancer, offering great promise to treat a broad spectrum of human solid and haematologic malignancies, especially where MYC is a driver.

cancer biology↗

bioRxiv2025

Fasting Rescues Locomotion in Neuromodulation-Deficient C. elegans via Octopamine-Gαq Signaling

Nutrient deprivation induces adaptive behavioral and physiological changes that are critical for survival. Here, we demonstrate that fasting ameliorates locomotion defects in Caenorhabditis elegans mutants lacking UNC-31/CAPS, a protein essential for dense-core vesicle (DCV)-mediated neuromodulation. Through forward genetic screening, we identified a gain-of-function mutation in egl-30, which encodes the heterotrimeric G protein subunit Gq, that suppresses the locomotion defects of unc-31 mutants under fed conditions. Transcriptomic analyses revealed that fasting induces upregulation of egl-30 and its downstream effectors in unc-31 mutants. Remarkably, exogenous octopamine treatment, which activates EGL-30/Gq signaling, mimicked the fasting response and restored locomotion in an EGL-30-dependent manner. Our findings uncover a mechanism of neuromodulatory plasticity, in which metabolic stress activates a compensatory octopamine-Gq signaling cascade to bypass impaired DCV-mediated neuromodulation, and suggest potential therapeutic strategies for CAPS-related neuropsychiatric disorders.

neuroscience↗

bioRxiv2025

Single and Multiple Buds Form characteristics in Amorphophallus muelleri

This study compared the physiological characteristics and transcriptomic profiles of single-budded and multi-budded corms, as well as developmental stages of foliar bulbils in Amorphophallus muelleri, to reveal regulatory mechanisms underlying bud development. Hormonal analysis showed that the IAA/CTK ratio was significantly higher in single-budded corms (1.9060 vs 1.5753), while multi-budded corms exhibited higher CTK (0.2766 g/g), BR (0.8692 p mol/g), and SLs (0.5090 ng/g) levels. During foliar bulbils development, ABA content peaked at 41.54 ng/g in the G2 stage, and sucrose reached maximum levels (6.86 mg/g) in the G1 stage. Transcriptomic sequencing identified 742 differentially expressed transcription factors, with over 80% of CRE1 and AHP genes in CTK signaling pathways significantly upregulated. KEGG pathway analysis highlighted starch/sucrose metabolism (215 DEGs) and plant hormone signal transduction (243 DEGs) as key regulatory networks. The study demonstrated that CTK signaling activation and carbohydrate metabolic reprogramming promoted multi-bud formation, while foliar bulbils development displayed stage-specific gene expression patterns, providing theoretical insights for improving konjac propagation coefficient.

physiology↗

bioRxiv2025

Targeting protein glycosylation and cholesterol metabolism in chemoresistant pancreatic cancer

Chemotherapy remains the cornerstone of pancreatic ductal adenocarcinoma (PDAC) treatment. However, most cases of PDAC finally progress to advanced chemoresistant disease. This highlights an urgent need to develop new pharmacological strategies to overcome chemotherapy resistance using clinical grade inhibitors. Here, we established a biobank comprising 260 organoid lines derived from 322 pancreatic cancer patients. These organoids underwent extensive multi-omics profiling and drug sensitivity testing for both chemotherapy and targeted therapy. Increased protein glycosylation and cholesterol metabolism signaling pathways were especially enriched in chemoresistant organoid lines. Importantly, chemoresistant PDAC organoids were effectively targeted with statins, inhibitors of cholesterol synthesis. Statins treatment attenuated protein glycosylation, cholesterol levels, and the epithelial-to-mesenchymal transition (EMT) signature in PDAC organoids. We further conducted a phase 2 clinical trial (NCT06241352) combining atorvastatin with chemotherapy in patients with advanced chemoresistant pancreatic cancer. Among 29 patients, 21 (72.4%) demonstrated a response, with tumor markers decreasing by more than 20%, suggesting durable responses and potential clinical benefits in this challenging patient population. ClinicalTrials.gov identifier: NCT06241352.

cancer biology↗

bioRxiv2025

Transferrin receptor 1 (TfR1) functions as an entry receptor for scale drop disease virus to invade host cell via clathrin-mediated endocytosis

Scale drop disease virus (SDDV), a distinct member of the genus Megalocytivirus within the Iridoviridae family, has emerged as a novel threat to global teleost aquaculture. Despite its importance, the pathogenic mechanism of SDDV remains largely elusive. In this study, we identified mandarin fish transferrin receptor 1 (mfTfR1) as an entry receptor for SDDV to invade host cells. Firstly, mfTfR1 was detected in high abundance in purified SDDV virions and exhibited dynamic responses to SDDV infection, showing distinct regulatory patterns both in vivo and in vitro. Overexpression of mfTfR1 in low-permissive fathead minnow (FHM) cells significantly enhanced SDDV replication, particularly during the early stages of viral binding and entry. Conversely, antibody-blocking experiments and treatment with the TfR1 inhibitor ferristatin II significantly suppressed SDDV entry. Further investigation revealed that mfTfR1 directly interacted with the major capsid protein (MCP) of SDDV, and the helical domain of mfTfR1 was identified as the crucial docking site. The binding site within the helical domain was determined, and disrupting this interaction significantly reduced viral entry and host mortality. Finally, we demonstrated that SDDV could activate Src kinase-mediated tyrosine phosphorylation of mfTfR1. This phosphorylation event enhanced the internalization of mfTfR1 and facilitated clathrin-mediated endocytosis. Collectively, our study provides compelling evidence to confirm that mfTfR1 functions as an entry receptor that mediates SDDV entry into host cells via clathrin-mediated endocytosis, leading to a lethal infection outcome. Our work lays the ground work for the development of targeted therapeutic strategies to mitigate the impact of SDDV in aquaculture. IMPORTANCETfR1, a dimeric glycoprotein classified as a type II transmembrane receptor, facilitates the cellular internalization of holo-transferrin. In several mammalian and avian RNA viruses, TfR1 serves as a crucial receptor to mediate the entry of viruses into host cells. As an emerging large DNA virus, SDDV poses an emerging threat to teleosts globally, however, its underlying pathogenic mechanisms remain poorly understood. In this study, we are the first to identify mfTfR1 as a crucial receptor for SDDV entry. We demonstrated a specific interaction between mfTfR1 and the major capsid protein (MCP) of SDDV, with the helical domain of mfTfR1 acting as the binding site. Moreover, we confirmed that SDDV enters cells through mfTfR1-mediated clathrin-dependent endocytosis. This work highlights the essential role of TfR1 in aquatic DNA viral infections and establishes the theoretical foundation for developing targeted therapeutic strategies against SDDV.

microbiology↗

bioRxiv2025

Munc13-4 mediates tumor immune evasion by regulating the sorting and secretion of PD-L1 via exosomes

Tumor-derived extracellular vesicles primarily carry PD-L1 via exosomes, which interact with PD-1 receptors on T cells, impacting immune responses in the tumor microenvironment and beyond, leading to a more extensive immunosuppressive landscape. However, the mechanisms governing exosomal PD-L1 sorting and secretion remain elusive. In this study, we identified Munc13-4 as a crucial regulator of exosomal PD-L1 sorting and secretion. Deletion of Munc13-4 in breast tumors enhances T cell-mediated anti-tumor immunity, suppresses tumor growth, and improves the efficacy of immune checkpoint inhibitors. Our results illustrate how Munc13-4 collaborates with HRS, Rab27, and SNAREs to facilitate PD-L1 sorting and secretion via exosomes. The cryo-EM structure of the Munc13-4-Rab27a complex provide new insights into its role in exosome secretion. Importantly, we discovered that Munc13-4 has a novel role in sorting PD-L1 onto exosomes, which relies on the formation of a ternary complex with PD-L1 and HRS. In addition, IFN{gamma} stimulation modifies Munc13-4 and HRS, establishing a dynamic regulatory mechanism that enables tumor cells to adapt to immune pressure by modulating PD-L1 sorting. Using a specially designed peptide to disrupt the Munc13-4-PD-L1 interaction and impede PD-L1 sorting significantly enhances anti-tumor immunity and slows tumor growth in vivo. These results highlight the potential of targeting the Munc13-4-PD-L1 axis to suppress tumor immune evasion.

cell biology↗

bioRxiv2025

Scale drop disease virus (SDDV) triggering ferroptosis both in vivo and in vitro facilitates virus infection via targeting transferrin receptor 1 (TfR1)

Scale drop disease virus (SDDV) is a distinct member in genus Megalocytivirus of family Iridoviridae, garnering increasing attention due to its significant threat to teleost. Ferroptosis is a new type of cell death discovered recently and involved in various viral infections. Knowledges on SDDV induced ferroptosis remains unclear. Here, we demonstrated that SDDV infection triggers ferroptosis, as evidenced by hallmark features such as iron overload, massive lipid peroxides accumulation, glutathione depletion and glutathione peroxidase 4 downregulation. SDDV-infected MFF-1 cells exhibited increased reactive oxygen species production and mitochondrial shrinkage. Treatment with Ferrostatin-1, a potent ferroptosis inhibitor, significantly attenuated SDDV replication in MFF-1 cells and could improve the survival of mandarin fish upon SDDV challenge. Treatment with an iron chelator mitigated ferroptosis and reduced the mortality of mandarin fish following SDDV infection, suggesting that SDDV-induced ferroptosis is iron-dependent. Finally, we demonstrated that SDDV infection could upregulate the expression of transferrin receptor protein 1 (TfR1), a critical iron transporter, to disrupt cellular iron homeostasis, induce ferroptosis, and then facilitate viral infection. Collectively, our findings provide compelling evidence that SDDV infection induces ferroptosis by targeting TfR1 to facilitate virus infection. Inhibiting ferroptosis represents a promising immunotherapeutic strategy for combating SDDV infection in aquaculture.

microbiology↗

bioRxiv2025

Expanding the Bacteroides synthetic biology toolkit to develop an in vivo intestinal malabsorption biosensor.

The human gut is a highly dynamic physical environment where perturbations--including factors such as acidification, oxygenation, and particle concentration (osmolality)--can influence microbiota composition and contribute to disease states. Understanding gut environmental changes is essential for advancing diagnostic and therapeutic strategies for gut health. However, non-invasive methods for continuous monitoring remain limited. The bacterial gut microbiota represents a powerful platform for continuous, non-invasive biosensing technologies for the gut environment, with genetically tractable commensal species like Bacteroides thetaiotaomicron (B. theta) emerging as promising hosts for engineered biosensors. However, the availability of genetic tools for precise, modular environmental sensing and reporting control in B. theta remains limited. Here, we present an expanded genetic engineering toolkit for B. theta that enables precise, fluorescence-based environmental sensing of the gut environment. This toolkit includes i) three libraries of orthogonally inducible promoters capable of driving fluorescence expression, ii) a DNA-based system to tune repressor activity in B. theta, iii) a resulting modular transcriptional reporter circuit that integrates native promoter activation with fluorescent outputs, and iv) characterization of a novel plasmid integration mode in B. theta. To demonstrate its utility, we engineered biosensors for gut malabsorption, a condition characterized by increased luminal osmolality. Using identified osmolality-responsive native promoters from B. theta, we made biosensors capable of detecting changes in gut physiology through graded fluorescent outputs. These biosensors were validated both in vitro and in vivo using a murine model of laxative-induced malabsorption, where they enabled continuous, long-term, non-invasive monitoring of single-cell response from fecal samples with sensitivity to subclinical malabsorption levels. By expanding the genetic toolkit for Bacteroides and demonstrating its use in a physiologically relevant context, this approach highlights the potential of engineered gut bacteria as a monitoring platform for diverse gut health applications. This work advances strategies for microbial biosensing and positions gut commensals as key players in next-generation diagnostic methods.

synthetic biology↗

bioRxiv2025

Ultrasound Directly Activates Sparse Neurons and Modulates Visual Circuits in Deafened Mice

Focused ultrasound neuromodulation (FUN) is widely regarded as a next-generation technology for neural modulation, with applications spanning rodents, non-human primates, and humans. While mechanistic studies are advancing, a persistent confound--auditory interference--casts doubt on whether ultrasound exerts direct mechanical effects or merely indirect auditory responses. To resolve this, we engineered a circular ultrasound transducer compatible with two-photon calcium imaging and examined its effects on the primary visual cortex (V1) of surgically deafened mice, eliminating auditory contributions. Our results reveal that ultrasound directly activates a sparse subset of ultrasound-sensitive neurons (USSN) (response rate >30%) in V1, comprising only a small fraction of the total population and exhibiting a spatially sparse distribution. The proportion of USSN scale with ultrasound pressure, confirming a direct neuromodulatory effect independent of audition. Intriguingly, despite this sparse activation, ultrasound significantly modulates V1 circuitry: it alters the dynamics of light-sensitive neurons (LSN), with subsets showing excitation, inhibition, or no change in response to visual stimuli. These findings provide the first rigorous in-vivo evidence that FUN induces direct mechanical effects on neural activity, disentangling them from auditory confounds. By demonstrating both the specificity and broader circuit-level impact of ultrasound in deafened mice, this study reframes our understanding of FUNs mechanisms and strengthens its potential as a precise neuromodulatory tool. HighlightsO_LIUltrasound directly activates sparse neurons in deafened mice, demonstrating auditory-independent effects. C_LIO_LISparse ultrasound-sensitive neurons in V1 show pressure-dependent responses. C_LIO_LIUltrasound modulates visual circuitry, with diverse excitatory and inhibitory effects. C_LI

neuroscience↗

bioRxiv2025

Hyperactivity of the Amygdala Mediates Depressive-Like Phenotypes and Decreased Serotonin Release

Clinical and preclinical studies have consistently demonstrated a correlation between hyperactivity of the amygdala and the onset of depression. However, the underlying mechanisms influencing serotonin levels a critical neurotransmitter implicated in depression and a primary target for selective serotonin reuptake inhibitors (SSRIs) remain inadequately understood. In this study, we employed a restrained inescapable shock (RIS) model to investigate these mechanisms in mice. The RIS paradigm elicited depressive-like phenotypes, increased c-Fos expression in the amygdala, diminished serotonin levels, and elevated corticosterone concentrations. Notably, chemogenetic inhibition of the amygdala mitigated depressive symptoms, reduced neuronal activity in this region, and restored serotonin levels. Anatomical analyses revealed a significant connectivity between the central amygdala (CeA) and the dorsal raphe nucleus (DRN). Fiber photometry recordings indicated that serotonergic neuronal activity in the DRN decreased in response to aversive stimuli, accompanied by amygdala activation following RIS, with no notable alterations in DRN GABAergic activity. These findings suggest that chronic stress may exacerbate amygdala hyperactivity, which subsequently inhibits serotonin release in the brain, potentially intensifying depressive states. Therefore, targeting amygdala hyperactivity may represent a novel therapeutic strategy for the management of stress-related depressive and anxiety disorders.

neuroscience↗

bioRxiv2025

Emergence of isochorismate-based salicylic acid biosynthesis within Brassicales

Salicylic acid (SA) is a major defense phytohormone. In Arabidopsis thaliana, the isochorismate (IC) pathway is the primary route for pathogen-induced SA biosynthesis. First, the IC synthase (ICS) catalyzes the isomerization of chorismate to IC in chloroplasts. Second, the chloroplast-localized MATE transporter EDS5 appears to transport IC from chloroplasts to the cytosol. Cytosolic IC is then further converted to SA via the GH3 amino acid-conjugating enzyme PBS3. While this pathway is genetically well-characterized in A. thaliana, its evolutionary origin and conservation remain controversial. In this study, through comprehensive phylogenetic, structural, and functional analyses, we demonstrate that the IC pathway emerged within the Brassicales order in a time span between the divergence of Carica papaya and Capparis spinosa. The evolution of the IC pathway was driven by three key adaptations during the time span: (1) enhancement of ICS activity, (2) neofunctionalization of EDS5 after duplication of its ancestral gene, and (3) evolution of a PBS3, whose activity is specialized for glutamate-conjugation to IC. Structural modeling and functional assays reveal that an enhanced salt bridge network in ICS enhanced its activity. One of the duplicated genes, EDS5, acquired key amino acid substitutions in the C-lobe, which likely contributed to the EDS5 neofunctionalization. In addition, the functional PBS3 clade, including A. thaliana PBS3, is restricted to a Brassicales clade. Taken together, this study addresses the evolutionary trajectory of IC-based SA biosynthesis.

plant biology↗

bioRxiv2025

Analysis of Head and Neck Cancer scRNA-seq Data Identified PRDM6 Promotes Tumor Progression by Modulating Immune Gene Expression

Head and neck squamous cell carcinoma (HNSCC) is a biologically aggressive and heterogeneous group of cancers with limited treatment options for patients who do not respond to standard therapies. While HPV-related HNSCCs tend to show better therapeutic outcomes, we still had limited understanding of the immune mechanisms underlying these cancers. Immune-responsive genes (IRGs) have emerged as critical factors in regulating both tumor progression and immune response. Recent advances in single-cell RNA sequencing (scRNA-seq) and the development of cell-type specific regulon inference tools, such as IRIS3, have provided new insights into the tumor immune microenvironment. In this study, we leveraged the IRIS3 platform to analyze scRNA-seq data from HNSCC patient samples, identifying novel transcription factor (TF)-IRG networks that contribute to tumor proliferation and immune escape. Specifically, we identified PRDM6, a histone methyltransferase, possesses the previously unknown role in promoting tumor cell proliferation by inducing IRG expression. We further demonstrated that HPV viral oncoproteins (E6/E7) oncoproteins up-regulate the PRDM6 expression, which associates PRDM6 with HPV-positive HNSCC.

cancer biology↗

bioRxiv2025

Synergistic neuroprotective and cognitive-enhancing effects of Walnut Peptide and Theanine in human brain organoid and mouse stress models

Stress is a prevalent mental health concern emerging predominantly in late adolescence or early adulthood. Since 2007, the Food and Drug Administration (FDA) has not approved any novel anxiolytic pharmaceuticals, fueling interest in nutritional supplements as alternative therapies for stress management. Building on prior zebrafish research, this study investigates the synergistic effects of Theanine (Th) and Walnut Peptide (WP) on stress mitigation and cognitive enhancement. Utilizing the human brain organoid stress (BO-stress) model, WP+ Th were observed to reduce stress and regulate expression of neurotransmitters, including Gamma-Aminobutyric Acid (GABA), serotonin (5-HT), dopamine (DA), and acetylcholine (Ach), as well as brain-derived neurotrophic factor (BDNF) and serotonin transporter (SERT). Subsequent in vivo study using C57BL/6J mouse-stress model demonstrated that the treatment (Th 85 mg/mL + WP 200 mg/mL), or administrated with vehicles, significantly improved their performance in stress and cognitive assessments, partially normalized neurotransmitter imbalances by modulating SERT and BDNF expression. These findings highlighted the potential of using WP + Th, particularly when delivered with vehicles (eg: powder/yogurt/milk), as a novel combined therapeutic approach for stress management and cognitive enhancement. We performed a correlation analysis between BO-stress model and mouse-stress model, revealing a alignement in the SERT levels.In addition, SERT was highly correlated with other markers in the mouse hippocampus and may represent a key target for modulating the balance between stress and cognition. Significance StatementThis study established an innovative human brain organoid-stress model and, in conjunction with mouse-stress model, elucidated the synergistic actions of Theanine and Walnut Peptide in mitigating stress and augmenting cognitive function. Their beneficial effects were mediated through the regulation of SERT and BDNF, presenting a promising non-pharmacological avenue for mental health care. In addition, species differences between humans and mice were also examined through correlation analysis of brain organoids and mouse models. Graphic abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=142 SRC="FIGDIR/small/640260v1_ufig1.gif" ALT="Figure 1"> View larger version (69K): org.highwire.dtl.DTLVardef@1a81886org.highwire.dtl.DTLVardef@e2281dorg.highwire.dtl.DTLVardef@1b7b167org.highwire.dtl.DTLVardef@dff9aa_HPS_FORMAT_FIGEXP M_FIG C_FIG

pharmacology and toxicology↗

bioRxiv2025

Construction and characterization of coronavirus nonstructural protein 3-host protein interaction networks unravel an important role of cleavage and polyadenylation specificity factor 6 in regulation of viral RNA replication

Coronavirus nonstructural protein 3 (nsp3) plays a crucial role in viral replication and immune evasion. However, functional and proteomic characterization of this protein, especially the interaction networks between nsp3 from different coronaviruses and host cell factors, is hindered by its huge size, complex structural feature and the presence of multiple transmembrane domains. In this study, we report the application of a high-performance cytoplasmic expression system to efficiently and accurately express the full-length nsp3 from betacoronavirus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and gammacoronavirus infectious bronchitis virus (IBV), to investigate their interactions with host proteins using proteomics approaches. Our study identified 1,150 host proteins that interact with IBV nsp3 and 920 with SARS-CoV-2 nsp3. Among them, 658 are shared by the two nsp3 proteins. Further validation and preliminary characterization of seven selected candidates, DDX5, DDX39, DHX9, elF4A3, SRRT and CPSF6, demonstrated the reproducibility and reliability of the proteomics data. More interestingly, an important regulatory role of the nsp3-CPSF6 interaction in the replication and transcription of IBV gRNA and sgRNA was unraveled. The construction of nsp3-host protein interaction networks from two distantly related coronaviruses would have provided a foundation for future studies of host cell factors in the regulation of coronavirus replication and pathogenesis.

microbiology↗