Dopamine (DA) is a pivotal neurotransmitter whose spatiotemporal dynamics are fundamentally linked to a myriad of neurological processes and disorders. Despite its significance, real-time visualization of DA across diverse cellular states remains challenging due to the scarcity of biocompatible and high-fidelity sensing strategies. Herein, we report a novel, trace Cu2+-catalyzed oxidative fluorogenic assay for the selective imaging of DA in living cells. By harnessing a trace copper-mediated redox cycle to catalyze DA oxidation, followed by specific fluorogenic recognition with 8-hydroxyjulolidine, this strategy circumvents the limitations of conventional oxidant-dependent approaches. The method yields a robust fluorescence response at 510 nm and maintains exceptional sensitivity and selectivity for DA over other catecholamines and biological interferents under physiological conditions. Notably, the strategy enabled the successful detection of endogenous DA dynamics in PC12 cells under normal, inflammatory, and depressive conditions. Time-lapse imaging revealed significantly suppressed DA release and altered DA reuptake kinetics in depressive cell models. This work provides a powerful analytical tool for elucidating the complex physiological and pathological roles of DA in neural systems.

This editorial highlights a study which examines immune checkpoint markers and inflammation indices in lymphoproliferative neoplasms in Egypt. A study by Sherief et al , published in World Journal of Clinical Oncology , present a novel combination of prognostic markers, showing that co-expression levels of programmed death-ligand 1/C-X-C motif chemokine receptor 3 (CXCR3) and programmed death-1 (PD-1)/CXCR3 were significantly elevated, particularly PD-1/CXCR3 in stage IV lymphoma (area under the curve = 0.959). The systemic inflammatory response index (SIRI) increased in late-stage patients (area under the curve = 0.846) but decreased after treatment (P < 0.001). The study challenges previous beliefs regarding SIRI and PD-1/CXCR3’s impact on survival rates. Importantly, SIRI relies only on routine blood test data, making it feasible for use in resource-limited settings. This research establishes an immune-inflammatory biomarker system for lymphoma in areas with high hepatitis C virus prevalence in Egypt and offers a low-cost, effective prognostic model for better patient stratification and improved cancer healthcare equity.

Sepsis is a life-threatening syndrome characterized by dysregulated host-immune responses, progressing through hyperinflammatory and immunosuppressive stages. Decoy receptor 3 (DcR3), a soluble member of the TNF receptor superfamily, serves as an immunomodulator in sepsis. Beyond neutralizing FasL, LIGHT, and TL1A to block apoptosis and inflammatory signaling, DcR3 regulates macrophage polarization, dendritic cell maturation, and immune cell survival through its heparan sulfate proteoglycan-binding domain. Evidence from cellular, molecular, and animal studies highlights its dual role in restoring immune balance by modulating both hyperinflammatory and immunosuppressive phases of sepsis. In this review, we summarize current evidence on DcR3 in sepsis and discuss translational challenges and future directions. Current rodent models lacking the TNFRSF6B gene are limited; however, transgenic mice expressing human DcR3 exhibit both protective and detrimental context-dependent effects. Translational challenges include the pharmacokinetics and immunogenicity of recombinant DcR3, although strategies such as PEGylation, nanoparticle encapsulation, and hydrogel delivery may improve its efficacy. Combining DcR3 with PD-1/PD-L1 inhibitors or immunometabolic agents like metformin and dimethyl itaconate presents promising therapeutic potential. Future research will focus on CRISPR/Cas9 knock-in mouse models, multi-omics mapping of DcR3 signaling, and biomarker-guided dosing. Although no DcR3-targeted clinical trials in sepsis have been conducted, DcR3 remains a precision-targeted immunotherapy with mechanistic and translational pathways; this review delineates key knowledge gaps that must be addressed to enable future clinical application.

Toxin-antitoxin (TA) systems contribute to bacterial stress adaptation and persistence, yet the structural and regulatory features of the VapBC5 module from Mycobacterium abscessus remain incompletely defined. Here, we characterize the VapBC5 toxin-antitoxin system using an integrated structural and functional approach. The VapBC5 complex was coexpressed, purified, crystallized, and its structure was determined at 2.24 Å resolution, revealing a heterotetrameric 2:2 assembly with a locally asymmetric interface. The C-terminal region of the antitoxin VapB5 engages the VapC5 toxin through an extensive network of hydrogen bonds and salt bridges, consistent with suppression of toxin activity. Structure-guided mutagenesis demonstrates that VapBC5 interface stability relies on a cooperative and distributed interaction network, and that disruption of multiple antitoxin-toxin contacts is required to unmask VapC5 activity in vivo. Functional assays in a heterologous Escherichia coli model show that VapC5 promotes antibiotic-tolerant survival under fluoroquinolone stress, whereas VapB5 counteracts this phenotype. Antitoxin-derived peptides were first screened in E. coli, and a selected peptide was subsequently evaluated in a laboratory M. abscessus strain, where it attenuated VapC5-associated persistence-related phenotypes. Together, these findings define the structural basis of VapBC5 regulation and support the use of antitoxin-derived peptides as molecular probes to modulate toxin-antitoxin-associated antibiotic tolerance in a mycobacterial context.

Oncolytic virus therapy (OVT) represents a promising frontier in cancer treatment. Despite its efficacy in clinical trials, variability in patient response, particularly resistance development, highlights the need for tailored therapeutic strategies. The Inositol Hexakisphosphate Kinase 2 (IP6K2) gene knock out was carried by CRISPR/Cas9 system. The evaluation of biomarkers of apoptosis and relevant pathways was conducted to be assessed. Attachment assay was conducted to verify the binding ability of virus to the host cells. Cell proliferation and apoptosis was assessed. Subcutaneous xenograft model was used to evaluate IP6K2 knock out influence in vivo. cBioPortal and TCGA database were applied to analyze genomic alterations in pan-cancer. IP6K2 was essential for effective Herpes Simplex Virus Type1 (HSV-1) replication and subsequent cell apoptosis, acting through the tumor Protein p53 (p53) and Cyclin-Dependent Kinase Inhibitor 1 A (p21) signaling axis. The tumor model demonstrated that tumors lacking IP6K2 exhibited resistance to HSV-1 oncolysis, resulting in diminished therapeutic outcomes. Analysis of cBioPortal and TCGA databases corroborated the potential resistance stemming from IP6K2 mutations across various cancer types, underscoring the necessity for pre-treatment IP6K2 status assessment. This study underscores the role of IP6K2 as potential markers of resistance, which opens avenues for precision medicine approaches in OVT. First reported that Inositol Hexakisphosphate Kinase 2 (IP6K2) is crucial for effective HSV-1 oncolytic virus therapy. CRISPR/Cas9-mediated knockout of IP6K2 resulted in decreased virus replication and apoptosis in cancer cells. Tumors lacking IP6K2 in vivo showed significant resistance to HSV-1 oncolysis, leading to reduced therapeutic efficacy. The p53-p21 signaling axis mediates the effects of IP6K2 on HSV-1 therapy efficacy. cBioPortal and TCGA database analyses confirmed that IP6K2 mutations are associated with resistance to oncolytic virus therapy across multiple cancer types. Highlighted the importance of assessing IP6K2 status prior to treatment to optimize oncolytic virus therapy for individual patients.

The cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) signaling pathway serves as a crucial component of the innate immune defense, playing a vital role in combating pathogen invasion. However, its dysregulation or abnormal activation can trigger the development of autoimmune diseases. This study demonstrated that Tanshinone IIA, a major lipid-soluble component of Salvia miltiorrhiza Bunge, can effectively inhibit the activation of the cGAS-STING signaling pathway. Mechanistically, Tanshinone IIA inhibits the transport of STING from the ER to the Golgi apparatus by weakening the interaction between STING and SEC24C, thereby preventing the activation of the cGAS-STING signaling pathway. Furthermore, Tanshinone IIA significantly ameliorated myocardial inflammation in WT and Trex1D18N/D18N mice. Our research indicates that Tanshinone IIA shows potential therapeutic value in alleviating autoimmune diseases by effectively inhibiting the abnormal activation of the cGAS-STING pathway.

BACKGROUND:Our investigation sought to uncover the intrinsic features of Head and Neck Squamous Cell Carcinoma (HNSCC), particularly the role of long non-coding RNAs implicated in disulfidptosis (DRLs). MATERIALS AND METHODS:We carried out lncRNA-mRNA RNA-Seq studies on HNSCC cells and harnessed the data from The Cancer Genome Atlas (TCGA), which includes 522 HNSCC tumors and 44 normal specimens. Bioinformatics evaluations aided in recognizing DRLs and estimating their prognostic value. Furthermore, we built a predictive model related to the chosen DRLs to scrutinize its linkage with the patients' prognosis. We also dug into tumor mutation loads and responses to chemotherapy. RESULTS:Our study identified three key DRLs (LINC02434, AC245041.2, and LINC02762) with considerable correlation to HNSCC prognosis. The risk model, utilizing these DRLs, successfully categorized patients into high-risk and low-risk clusters, uncovering differential survival trajectories. Moreover, the same risk model conveyed unique prognostic potential in HNSCC. Surveying the tumor microenvironment unfolded disparities between the groups, hinting toward potential implications for tactics in immunotherapy. We recognized distinct chemotherapeutic drugs with fluctuating responses across the risk clusters and molecular categories. CONCLUSION:This investigation not only sheds light on prospective therapeutic pathways but also enhances our grasp of the molecular intricacies of HNSCC.

Advances in synthetic biology have enabled innovative strategies for cancer therapy, yet precise control of therapeutic expression and biosafety remain critical challenges. To address these issues, a bacterial hybrid biorobot is developed using Escherichia coli (MG1655) engineered for localized activation by microwaves. Upon activation, the biorobot expresses glucose oxidase (GOx) at tumor sites, leading to glucose depletion and hydrogen peroxide generation. Surface-attached Cu2O nanoparticles catalyze this hydrogen peroxide through a Fenton-like reaction, producing reactive oxygen species that drive multiple forms of tumor cell death, including apoptosis, ferroptosis, and cuproptosis. Comprehensive in vitro and in vivo studies confirm the efficacy of this approach, while transcriptomic analysis reveals disruption of glucose metabolism and robust activation of antitumor immune responses. This work demonstrates the potential of this engineered bacterial platform as a safe and versatile tool for precise, multimodal cancer treatment.

The innate immune system relies on a variety of pathogen recognition receptors (PRRs) as the first line of defense against pathogenic invasions. Viruses have evolved multiple strategies to evade the host immune system through coevolution with hosts. The CRISPR-Cas system is an adaptive immune system in bacteria or archaea that defends against viral reinvasion by targeting nucleic acids for cleavage. Based on the characteristics of Cas proteins and their variants, the CRISPR-Cas system has been developed into a versatile gene-editing tool capable of gene knockout or knock-in operations to achieve genetic variations in organisms. It is now widely used in the study of viral immune evasion mechanisms. This chapter will introduce the use of the CRISPR-Cas9 system for editing herpes simplex virus 1 (HSV-1) genes to explore the mechanisms by which HSV-1 evades host innate immunity and the experimental procedures involved.

Objective: Combining immune checkpoint inhibitors and antiangiogenic agents offers a promising strategy to counteract the cooperative promotion of solid tumor growth by immune checkpoints and intratumoral angiogenesis. Methods: We investigated the potential of thalidomide (THD) and anti-PD-1 antibody (PD-1 mAb) in suppressing tumor growth, enhancing immunity, and inhibiting angiogenesis. Results: THD exhibited regulatory effects on PD-1 in CD4+ T cells and PD-L1 in cancer cells, along with tumor growth inhibition in A549 and Lewis lung carcinoma (LLC) cell lines. Combined with PD-1 mAb, THD increased intracellular IL-2 and IFN-γ expression in CD4+ T cells, enhanced granzyme (Gzm-B) expression in peripheral blood mononuclear cells (PBMCs), and reduced TNF-α expression in CD4+ T cells. In C57BL/6 mice, THD plus PD-1 mAb decreased LLC-derived lung tumor weight and volume, boosted CD8+ T cell infiltration in tumors, and reduced CD34+ intratumoral microvessel density Conclusion: This study highlights THD’s role in modifying the tumor microenvironment to enhance PD-1 mAb efficacy, proposing a clinically feasible approach for improving PD-1 mAb treatment outcomes.

Per- and polyfluoroalkyl substances (PFAS) are emerging persistent environmental pollutants with potential risks to microbial ecosystems. However, the influence of PFAS with different chain lengths on horizontal gene transfer, particularly plasmid-mediated antibiotic resistance genes (ARGs) conjugation, remains poorly understood. This study investigated the impacts of short-chain (PFBA, PFHxA) and long-chain (PFNA) PFAS exhibited dual effects on the conjugative transfer of ARGs: PFAS with lower concentration (<0.05 mg/L) enhanced ARGs transfer by increasing the permeability of the cell membrane and ROS content, while higher concentration (>0.05 mg/L) of PFAS led to stronger inhibition through suppressing adenosine triphosphate (ATP) production. The scarcity of ATP caused cells to rebuild their energy allocation strategies, diverting more energy towards maintaining vital life activities rather than for gene transmission. Notably, short-chain PFAS (e.g. PFBA) of smaller molecular possessed greater facility for entering cells and caused stronger dual effects on cells. However, long-chain PFAS with high hydrophobicity prefers to embed in the phospholipid bilayer, causing weaker dual effects on cells and less frequency of conjugative transfer. These findings revealed the distinct effects of PFAS with different chain lengths on the conjugative transfer of ARGs, and highlighted the critical role of the cell membrane in this phenomenon. This research provides critical insights into the ecological risks posed by PFAS.

The application of CRISPR-mediated library screening has fundamentally transformed functional genomics by revealing the complexity of virus-host interactions. This protocol describes the use of CRISPR-mediated library screening to identify key functional genes regulating the innate immune response to PEDV infection. We detail a step-by-step process, starting from the design and construction of a customized CRISPR knockout library targeting genes involved in innate immunity to the effective delivery of these constructs into cells using lentiviral vectors. Subsequently, we outline the process of identifying functional genes postviral attack, including the use of next-generation sequencing (NGS), to analyze and identify knockout cells that exhibit altered responses to infection. This integrated approach provides researchers in immunology and virology with a resource and a robust framework for uncovering the genetic basis of host-pathogen interactions and the arsenal of the innate immune system against viral invasions.

Lung and colorectal cancers are among the most devastating malignancies globally, contributing to millions of cancer-related deaths each year. Despite significant advancements in targeted therapies and immunotherapies, clinical challenges continue to arise. For example, Tyrosine Kinase Inhibitors (TKIs) used for treating Epidermal Growth Factor Receptor (EGFR)-mutant lung cancer often led to acquired resistance within 9 to 14 months. Meanwhile, immune checkpoint inhibitors, such as anti-PD-1/PD-L1 antibodies, achieve Objective Response Rates (ORR) of less than 30% in many patients. Additionally, the “don’t-eat-me” signal, mediated by the CD47 signal regulatory protein α (SIRPα) axis, allows tumour cells to evade macrophage phagocytosis, further diminishing the effectiveness of existing treatments. In this context, Antibody-Drug Conjugates (ADCs) which combine the specificity of monoclonal antibodies (mAbs) with the cytotoxicity of small-molecule drugs have emerged as a transformative strategy to address these limitations. The recent study by Chiang et al. (2025) on “7DC-DM1,” a non-cleavable CD47-targeting ADC, published in the International Journal of Biological Macromolecules, represents a significant advance in meeting the unmet needs of lung and colorectal cancer therapy. It offers new insights into ADC design, target engagement, and clinical translation. Furthermore, this editorial discusses potential application scenarios and future development directions for various types of ADCs.

Colorectal cancer is the second leading cause of cancer-related deaths following lung cancer in recent years. Therefore, lung or colorectal cancer therapy is very important for reducing mortality. In this study, we developed and characterized CD47-specific antibody-drug conjugates, namely 7DC-DM1 ADCs, to evaluate their therapeutic effects on lung and colorectal cancer. Both 7DC2-DM1 and 7DC4-DM1 demonstrated good binding affinities of 0.56 nM and 0.49 nM, respectively, and exhibited significant cytotoxicity, though they displayed different penetration effects. These findings suggest that the binding complexes of 7DC2-DM1 and 7DC4-DM1 with CD47 receptors adopt different conformations, leading to variations in their cellular internalized efficiencies. Molecular docking simulations revealed that 7DC2 and 7DC4 bind to CD47 molecules in distinct orientations and epitopes, differing between conserved and non-conserved regions. Furthermore, treatments with 7DC2-DM1 and 7DC4-DM1 displayed notable differences in antitumor effects in murine syngeneic tumor models derived from the MC38 cell line in C57BL/6 mice. In the tumor model treated with 7DC4-DM1, immunofluorescence staining analysis revealed a large area of necrosis in the tumor stroma, accompanied by a significant infiltration of CD11b-expressing immune cells. In summary, these results indicate that 7DC4-DM1 holds promise as a therapeutic agent for colorectal cancer treatment.

tRNA-derived small RNAs (tsRNAs), as a class of regulatory small noncoding RNA, have been implicated in a wide variety of human diseases. Large amounts of tsRNA-disease associations have been identified in recent years from accumulating studies. However, repositories for cataloging the detailed information on tsRNA-disease associations are scarce. In this study, we provide a tsRNADisease database by integrating experimentally and computationally supported tsRNA-disease associations from manual curation of literatures and other related resources. tsRNADisease contains 5571 manually curated associations between 4759 tsRNAs and 166 diseases with experimental evidence from 346 studies. In addition, it also contains 5013 predicted associations between 1297 tsRNAs and 111 diseases. tsRNADisease provides a user-friendly interface to browse, retrieve, and download data conveniently. This database can improve our understanding of tsRNA deregulation in diseases and serve as a valuable resource for investigating the mechanism of disease-related tsRNAs. tsRNADisease is freely available at http://www.compgenelab.info/tsRNADisease.

Transmissible gastroenteritis virus (TGEV) represents a significant threat to global swine production. In the absence of effective antiviral therapies, control relies primarily on vaccination. To identify potential therapeutic targets, we performed a genome-wide CRISPR/Cas9 screen in porcine IPEC-J2 cells, which revealed asparagine-linked glycosylation 5 (ALG5), asparagine-linked glycosylation 6 (ALG6), neurofibromin 2 (NF2), and fucosyltransferase 8 (FUT8) as essential host factors for TGEV infection. Functional characterization demonstrated that ALG5, ALG6, and NF2 knockout impaired viral adsorption and internalization through disruption of aminopeptidase N (pAPN) transcription or N-glycosylation. Consistently, tunicamycin-mediated inhibition of N-glycosylation suppressed TGEV infection. In contrast, FUT8 knockout specifically affects viral internalization and early replication by preventing the formation of double-membrane vesicles (DMVs) but does not affect pAPN expression. This role was independent of FUT8's fucosyltransferase activity, as the enzymatic inhibitor FDW028 had no effect. Mechanistically, we found that FUT8 interacts with the TGEV nonstructural proteins NSP3 and NSP4 to facilitate DMV biogenesis. Our findings delineate distinct mechanisms by which host factors support TGEV infection and provide novel insights for the development of targeted antiviral strategies.

MicroRNAs (miRNAs) play key roles in development and disease, and have great biomarker potential. However, because miRNA expression is highly cell-type specific, identifying miRNA biomarkers from complex tissues is hampered by the underlying cell-type heterogeneity. Due to that current single-cell RNA-Seq protocols are lagging behind for quantification of miRNA expression, and most miRNA profiling samples do not have matched mRNA expression or DNA methylation data for cell-type deconvolution, it is an urgent need to develop computational methods for cell-type proportion estimation of bulk-tissue miRNA data. Here we present a novel miRNA expression reference library and deconvolution tool for cell-type composition estimation of complex tissues. We show that our tool is accurate and robust for deconvolution in whole blood as well as in different solid tissues. By applying this tool to a range of different biological contexts, we demonstrate its value for screening of age-associated miRNAs, for monitoring the immune landscape in infectious diseases like COVID-19, as well as for identifying cell-type-specific miRNA biomarkers for early diagnosis and prognosis of human cancers. Our work establishes a computational framework for accurate cell-type mixture deconvolution of miRNA data.

[This corrects the article DOI: 10.3389/fimmu.2025.1561563.].

Background:Quasipaa spinosa crude extract (QSce), a natural source rich in proteins such as parvalbumin (PV), has been traditionally used to promote physical recovery. However, its mechanisms in mitigating exercise-induced fatigue remain unclear. Methods: Using a murine treadmill exhaustion model, we evaluated the effects of QS-derived Parvalbumin (QsPV) (30 and 150 mg/kg/day) on endurance capacity, oxidative stress, tissue injury, and muscle function. Indicators measured included time to exhaustion, intracellular calcium levels, antioxidant enzymes [superoxide dismutase (SOD), glutathione peroxidase (GSH-Px)], lipid peroxidation (malondialdehyde, MDA), injury markers [creatine kinase (CK), lactate dehydrogenase (LDH), cardiac troponin I (cTnI)], renal function (blood urea), and muscle force. Results: QsPV-150 significantly increased time to exhaustion by 34.6% compared to the exercise-only group (p < 0.01). It reduced MDA by 41.2% in skeletal muscle and increased SOD and GSH-Px levels by 35.4% and 28.1%, respectively. Serum CK, LDH, and cTnI were reduced by 39.5%, 31.7%, and 26.8%, respectively, indicating protection against muscle and cardiac injury. QsPV also decreased blood urea by 22.3% and improved renal histology, with reduced glomerular damage and tubular lesions. At the molecular level, QsPV restored calcium balance and downregulated calpain-1/2 and atrophy-related genes (MuRF-1, MAFbx-32). Muscle contractile force (GAS and SOL) improved by 12.2-20.3%. Conclusions: QsPV attenuates exercise-induced fatigue through multi-organ protection involving calcium buffering, oxidative stress reduction, and anti-atrophy effects. These findings support its potential as a natural recovery-enhancing supplement, pending further clinical and pharmacokinetic studies.