Regulated cell death (RCD) is well-known as a controlled form of cell death regulated by one or more cascading signaling pathways. Over the past few decades, increasing evidence has implicated various non-apoptotic forms of RCD in neurons—including ferroptosis, parthanatos, necroptosis, pyroptosis, autophagic cell death, paraptosis, and cuproptosis—in the pathogenesis of neurodegenerative diseases (NDs) and their associated clinical manifestations. We provide an in-depth analysis of the associations between these RCDs and NDs, including Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), and multiple sclerosis (MS), and highlight the potential of modulating non-apoptotic RCD subtypes as neuroprotective targets. Besides, we highlight the crosstalk mechanisms among different non-apoptotic RCDs in NDs and the key targets regulating the crosstalk, which hold significant promise for developing dual-functional inhibitors that precisely modulate the pathological microenvironment and overcome drug resistance. As our understanding of death signaling networks deepens, such strategies may lead to breakthrough therapies for multiple NDs. Moreover, we further discuss the emerging small molecule compounds targeting non-apoptotic RCDs and their current research progress in clinical trials for the treatment of NDs, which may provide novel directions for related drugs. This comprehensive analysis paves the way for future research and therapeutic strategies aimed at harnessing non-apoptotic RCD pathways to mitigate neurodegeneration and improve patient outcomes.

Parkinson's disease (PD) is a progressive neurodegenerative disorder characterized by the loss of dopaminergic (DA) neurons and the pathological aggregation of α-synuclein. While current pharmacological therapies provide symptomatic relief, they do not halt or reverse disease progression. Cell-based regenerative strategies have emerged as promising approaches to restore DA function and target the complex, multifactorial pathophysiology of PD. This review critically examines current approaches, including transplantation of fetal ventral mesencephalic tissue, pluripotent stem cell-derived midbrain DA progenitors, and in vivo reprogramming of endogenous cells. In addition, supportive cell types, such as mesenchymal stromal cells and carotid body glomus cells, provide neuroprotective and immunomodulatory effects via paracrine signaling. We summarize preclinical and clinical evidence on graft survival, integration, and functional recovery, and discuss key determinants of therapeutic efficacy, including mitochondrial function and bioenergetic integrity, immune compatibility, and biomaterial scaffolding. Despite significant progress, major challenges remain regarding long-term efficacy, graft standardization, and host-graft interactions. Ongoing translational advances are poised to drive the development of disease-modifying cell therapies capable of delivering durable clinical benefits and improving long-term outcomes in PD.

Obesity and diabetes are chronic metabolic diseases affecting millions worldwide. Current treatments, including lifestyle changes, medications, and surgery, face challenges like poor adherence and side effects. Glucagon-like peptide-1 receptor agonists (GLP-1RAs) are recommended innovative medications for these conditions, with studies showing significant clinical benefits. GLP-1RAs are traditionally delivered orally or via subcutaneous injections, but these methods have limitations, including low bioavailability, poor solubility, the need for high doses, gastrointestinal side effects, and frequent dosing requirements. Novel delivery technologies offer promising strategies to overcome these challenges and enhance therapeutic effectiveness. Recent advances in drug delivery technologies, including nanocarrier- and microcarrier-based systems, hydrogels, microneedles, and innovative formulations such as long-acting, co- and/or nano-formulated agents, offer promising strategies to enhance the delivery, efficacy, and patient adherence of GLP-1RAs for obesity and diabetes. This review focuses on innovative delivery technologies developed for three main GLP-1RAs: exenatide, liraglutide, and semaglutide. We present a review of advancements in drug delivery systems, exploring technologies employed in the development of these agents, as well as future challenges. It is crucial to note that these technologies are still in early development, and further studies are needed to confirm their long-term safety, efficacy, and cost-effectiveness in clinical use.

Degenerative diseases are a group of medical conditions characterized by the progressive and irreversible deterioration of cells, tissues, and organs over time. Emerging evidence highlights the alteration and functions of the gut microbiome in the development of degenerative diseases. Ferroptosis, a regulated form of cell death characterized by iron-dependent lipid peroxidation, has been implicated as a pivotal factor in the regulatory effect of the gut microbiome on degenerative diseases. Moreover, gut metabolites, particularly short-chain fatty acids and trimethylamine N-oxide, are closely related to iron overload, redox imbalance, and lipid peroxidation. Recently, microbiome-based therapies, such as fecal microbiota transplantation, have been considered novel therapeutic strategies. In this review, we focus on degenerative diseases and explore the interactions between the gut microbiome and ferroptosis, aiming to provide new insights into the underlying mechanisms and clinical implications.

African swine fever virus (ASFV) causes a highly contagious and lethal disease in domestic and wild pigs, posing a significant threat to global swine production. The lack of effective vaccines or antiviral therapies underscores the urgent need for alternative intervention strategies. In recent years, notable progress has been made in developing antiviral agents that target both viral components and host-dependent pathways. This review provides a comprehensive summary of recent advances in ASFV antiviral research, with a focus on therapeutic interventions aimed at viral-encoded proteins and host factors, particularly small-molecule inhibitors and their mechanisms of action. In addition, the review highlights target-agnostic and multifaceted antiviral strategies, including physical inactivation, early life cycle disruption, and gene-level interference, which act through broad or partially understood mechanisms. By integrating mechanistic insights with emerging therapeutic approaches, this review supports the rational design of antiviral interventions against ASFV and informs the subsequent exploration of candidate drugs with clinical translational potential.

Phase separation of biological macromolecules is a ubiquitous cellular mechanism for concentrating and compartmentalizing biochemical reactions. Emerging evidence reveals that biomolecular condensates formed via liquid-liquid phase separation (LLPS) play integral roles in diverse physiological processes, including gene expression and intracellular signaling, enabling rapid and delicate cellular responses. Dysregulation of phase separation is increasingly implicated in the pathogenesis of major diseases, ranging from neurodegenerative disorders and cancers to viral infections and aging. Consequently, deciphering the molecular mechanisms governing LLPS and developing strategies for its pharmacological modulation, particularly via small molecules, represent promising therapeutic targets offering novel approaches for disease intervention. In this review, we provide a concise overview of biomolecular condensates formation and their functions in diseases regulation. Mainly, we catalog commonly employed tool compounds, reported small-molecule modulators of LLPS, and related clinical progress within this rapidly evolving research area. Furthermore, we integrate recent technological breakthroughs in LLPS to envision the future trajectory and therapeutic potential of this field.

The combination of cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) signaling pathway modulation with nanotechnology offers a promising strategy for the development of more effective and less toxic therapies. This review summarizes the latest clinical progress of STING agonists and inhibitors, with a particular focus on the role of nanomaterials in regulating the cGAS-STING pathway across a range of diseases. In oncology, STING activation enhances anti-tumor immunity by stimulating immune cells, while nanocarriers improve the stability and targeting precision of STING agonists, facilitating synergistic effects with other immunotherapies. In inflammatory and autoimmune diseases, regulating STING activation helps alleviate the production of excessive pro-inflammatory cytokines, restore immune homeostasis, and prevent tissue damage. Nanomaterials, such as cell-derived membranes, further enhance targeted delivery and biocompatibility, addressing key limitations of existing treatment strategies. What distinguishes this review is an in-depth analysis of the current clinical progress of STING agonists and inhibitors, providing a comprehensive overview of both ongoing clinical trials and preclinical advancements. We also critically evaluate the specific challenges encountered in translating STING nanomaterials into clinical practice. These challenges present significant barriers to the widespread application of STING-based therapies, underscoring the need for further optimization to realize their full potential.

Inflammation-related diseases account for over 50% of global disease-associated mortality; the core pathological mechanisms of these diseases are closely linked to functional dysregulation of immune cells such as macrophages and T cells. Aberrantly activated immune cells excessively secrete inflammatory mediators, which drive chronic inflammatory cascades and trigger irreversible tissue damage. In recent years, immune cell-based therapeutic agents (ICTAs) have garnered significant attention due to their inherent targeting specificity and immunomodulatory capabilities, encompassing whole immune cells, cell membranes, or extracellular vesicles serving as active therapeutics or delivery carriers. This review systematically elaborates on strategies for constructing ICTAs through nanoengineering, genetic engineering, and membrane-fused engineering, while outlining their integrating applications with other delivery devices. Furthermore, we summarize the preclinical and clinical trial advancements of ICTAs in various diseases such as tumors, rheumatoid arthritis, diabetes, atherosclerosis, Alzheimer's disease, inflammatory bowel disease, ischemia/reperfusion injury, sepsis, and hemophagocytic lymphohistiocytosis. These insights establish an interdisciplinary design framework for developing clinically applicable ICTAs and propose novel therapeutic approaches for inflammation-related diseases.

Ischemic stroke (IS) remains a leading cause of global death and disability, with treatment effectiveness limited by its complex mechanisms, which include blood-brain barrier (BBB) disruption, neuroinflammation, excitotoxicity, oxidative stress, and cell death. This review summarizes emerging research on extracellular vesicle (EV)-based therapies for IS. EVs, sourced from animals, plants, and microbes, have unique benefits as natural nanocarriers, such as inherent BBB permeability, biocompatibility, and the ability to deliver multiple therapeutic cargos (like miRNAs, proteins, and drugs). We evaluate how EVs target key IS issues: (1) restoring BBB integrity by stabilizing tight junctions (TJs) and reducing matrix metalloproteinases (MMPs), (2) modulating microglia to reduce neuroinflammation, (3) decreasing excitotoxicity, (4) scavenging reactive oxygen species (ROS) to lessen oxidative stress, and (5) inhibiting apoptosis, ferroptosis, and other cell death pathways. Additionally, engineered EVs, such as antibody-conjugated or magnetically guided types, exhibit improved targeting and treatment accuracy, yielding promising results. Despite significant preclinical promise, clinical application faces challenges in standardization, scalable production, and delivery improvement. EVs offer a transformative, multi-targeted approach with the potential to overcome current limitations in IS treatment.

Intratumoural bacteria have been shown to play conflicting roles within tumours, especially in the modulation of immune responses. Moreover, bacteria have been linked to a variety of cancer types and have different prognostic values depending on the cancer. This heterogeneity is not only shaped by tumour molecular subtypes, spatial location, cancer progression stage, genetic alterations, and the presence of multiple subclones, but is also influenced by variations in angiogenesis, oxygen levels, microbial sources, endocytosis and micropinocytosis. In this review, we describe the diversity of intratumoural bacteria across 25 cancer types covering an extensive spectrum of analyzed intratumoural bacteria. Furthermore, the dual roles and mechanisms of their involvement in tumour progression and anticancer activity are summarized. Additionally, interventions and applications of engineered bacteria in cancer therapy, especially strategies used in clinical trials, are illustrated. This work describes the roles of intratumoural bacteria and their metabolic byproducts as regulators within the tumour microenvironment and highlights the potential translation of bacteria for cancer therapy.

Pathogenic mutations within protein-coding regions of genomic DNA can disrupt protein structure and lead to hereditary disorders. Genome-editing technologies, particularly those based on clustered, regularly interspaced, short palindromic repeats-associated protein (CRISPR-Cas), are promising therapeutic tools for correcting genetic abnormalities. To date, viral delivery vectors for genome-editing biomacromolecules have shown numerous promises in treating genetic disorders. However, safe viral delivery for genome-editing components remains challenging, largely due to the immunogenicity of viruses. As an alternative, non-viral delivery systems are emerging as a safer choice and may offer solutions to address the safety challenges. In this review, we first introduce CRISPR-Cas9-based genome editing tools and their delivery formats. Then, we outline the pathology of major genetic disorders and both preclinical and clinical approaches for these diseases by therapeutic genome editing, and provide an overview of current non-viral delivery strategies and their potential to overcome existing limitations. Finally, we discuss the current challenges and future outlooks of non-viral delivery of gene-editing components in treating genetic diseases.

Medicinal plants synthesize an immense diversity of specialized metabolites that play crucial roles in ecological interactions and serve as valuable pharmaceutical resources. However, the biosynthetic pathways responsible for this chemical diversity remain largely uncharacterized. These pathways are often complex, involving multiple steps that are spatially and temporally orchestrated within highly specialized or rare cell types. Classical bulk omics approaches obscure such cellular heterogeneity by averaging signals across tissues, limiting their utility in resolving cell-specific metabolic processes. Recent advances in single-cell and spatial omics technologies have revolutionized the ability to investigate plant metabolism at high spatiotemporal resolution, as exemplified by monoterpene indole alkaloids and Taxol biosynthesis. In this review, we highlight key technological advances in plant single-cell and spatial omics, examine their applications in pathway discovery and partitioning, and discuss emerging directions for harnessing these tools in plant synthetic biology and metabolic engineering. These developments promise to accelerate the systemic mapping of plant metabolic networks and facilitate their biotechnological exploitation for pharmaceutical development.

Accurate prediction of drug-induced gene expression profiles is crucial for phenotype-based drug discovery. Although computational methods have shown potential, they struggle with the complexities of varying doses and durations. To overcome these limitations, we developed DeepICER, a model that predicts gene expression profiles induced by chemical perturbations across any dose and duration. Utilizing a bilinear attention mechanism, DeepICER captures the interplay between dose, duration, and basal gene expression, enabling accurate predictions for novel compounds and cell lines. DeepICER outperforms existing models with superior flexibility in handling any dose and duration and accuracy, achieving a 45.1% improvement in predictive performance. Experimental validation confirmed that PD-166285, identified by DeepICER, exhibits stronger inhibitory effects on A549 cells compared to paclitaxel. To enhance accessibility, DeepICER is developed as an online platform, providing researchers with a tool to predict gene expression in compound-treated cells, thereby advancing drug repurposing and accelerating drug discovery.

Previous studies have shown that heat shock protein 90 (Hsp90) inhibitors can reduce seizures in temporal lobe epilepsy (TLE) by upregulating excitatory amino acid transporter 2 (EAAT2, also known as GLT-1). While the Hsp90 inhibitor 17-AAG is effective, its long-term use raises toxicity concerns. This study aimed to identify a safer Hsp90 inhibitor by screening benzenoid ansamycin derivatives for higher binding affinity and lower toxicity. Among nine natural benzenoid ansamycins and their derivatives screened, reblastatin emerged as the top candidate, exhibiting the highest binding affinity to Hsp90. Compared to geldanamycin and 17-AAG, reblastatin demonstrated significantly lower cytotoxicity in HEK293 and HepG2 cells. Like 17-AAG, reblastatin upregulated EAAT2 levels by disrupting the association among Hsp90, EAAT2, and the 20S proteasome. In a kainic acid-induced TLE mouse model, reblastatin reduced seizure frequency by 50%, with long-term treatment showing toxicity comparable to vehicle controls. Additionally, behavioral tests revealed neuroprotective effects of reblastatin in mouse models of Alzheimer's disease and Parkinson's disease. These findings collectively suggest that reblastatin is a promising Hsp90 inhibitor for treating TLE and excitotoxic conditions associated with neurodegenerative diseases.

Doxorubicin (DOX)-induced cardiotoxicity (DIC) is a major health threat that limits its clinical application. While mitochondrial dysfunction, oxidative stress and ferroptosis are implicated in DIC pathology, the precise mechanism remains elusive. This study evaluated the role of cluster of differentiation 74 (CD74), an immunoregulatory protein, in DIC. Our findings revealed elevated CD74 levels in blood samples from DOX-exposed patients and DOX-challenged mouse hearts. CD74 deletion mitigated DOX-indued cardiac remodeling, contractile anomaly, mitochondrial abnormalities, apoptosis, and ferroptosis. Mechanistically, CD74 bound to DNA synthesis molecule ribonucleotide reductase M2 (RRM2), redistributing it from cytoplasm to plasma membrane, impairing DOX-induced repair and exacerbating mitochondrial injury, apoptosis, and ferroptosis via activation of RRM2/p53 cascade. Notably, a CD74 mutant (aa 220-250) failed to aggravate DOX-induced cardiac dysfunction, unlike WT CD74. Moreover, the protective effects of CD74 inhibition in cardiomyocytes were negated by p53 activation, highlighting its role in DOX-induced damage. Treatment with CD74 inhibitor Amifostine in a DIC mouse model significantly alleviated cardiac remodeling and functional impairment by reducing oxidative stress and ferroptosis. Transwell study using the CD74-null Raw 264.7 macrophages and cardiomyocytes revealed that CD74 knockdown in macrophages overly attenuated DOX-instigated cardiomyocyte dysfunction. These findings establish CD74 as a potential therapeutic target for DIC, as its regulation of RRM2 cytomembrane diversion and ferroptosis ultimately drives cardiac remodeling and contractile anomalies in response to DOX challenge.

TET2-mediated clonal hematopoiesis of indeterminate potential (CHIP) is a known cardiovascular risk factor, but its role in cardiac aging and potential for pharmacological intervention remain unclear. Herein, Mendelian randomization using large-scale genome-wide association studies (GWAS) data assessed CHIP's causal impact on aging and cardiovascular disease that revealed significant causal associations between TET2-CHIP, CVD, and aging biomarkers. Transcriptome-guided screening identified oridonin as a candidate compound reversing CHIP- and aging-associated gene signatures. Multi-tiered target prediction combining chemical structure-based algorithms and transcriptomic correlation identified KDM5C as a key target, validated by enzymatic inhibition and surface plasmon resonance assays. In vivo and in vitro administration of oridonin significantly ameliorated cardiac dysfunction and pathological remodeling in the CHIP model. Epigenetic regulation was profiled via ChIP-seq and RNA-seq, focusing on H3K4me3-mediated transcription. Tet2⁺/⁻BMT mice exhibited age-progressive myocardial fibrosis, inflammation, senescence, and functional decline. Mechanistically, oridonin inhibited KDM5C histone demethylase, restored H3K4me3 levels, and activated the SIRT2 anti-aging pathway. Rescue experiments using gene overexpression and recombinant protein supplementation confirmed the functional role of the KDM5C-H3K4me3-SIRT2-S100A8 axis in mediating oridonin's effects. Overall, TET2-driven CHIP promotes cardiac aging, as evidenced by human genetic analyses and long-term BMT models. Oridonin, by inhibiting KDM5C and restoring H3K4me3-dependent SIRT2 signaling, mitigates CHIP-induced myocardial aging.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is the most common chronic liver disease, posing public health risks from potential irreversible liver damage. Curcumin (Cur), a polyphenolic compound from Zingiberaceae and Araceae, exhibits antihyperlipidemic and insulin-sensitizing effects. However, its targets and mechanisms in MASLD remain unclear. This study aimed to identify the potential targets and mechanisms by which Cur ameliorates MASLD. Cur markedly improved metabolic disorders and inflammation in rats and reduced intracellular lipid accumulation in HepG2 cells, primary hepatocytes, and AML12 cells. TKFC was identified as a direct target of Cur which binds to TKFC at Val136 and Glu538, thereby activating it. Gene microarray screening showed that Cur down-regulated the mRNA expression of Gpat3. TKFC knockdown attenuated the down-regulation of GPAT3 by Cur, whereas TKFC overexpression reversed this effect. In Tkfc-deficient mice, the therapeutic effects of Cur were attenuated and the down-regulation of GPAT3 and related proteins was hindered, thereby promoting triglyceride production in the liver. Further mechanistic studies revealed that Cur targets TKFC to inhibit GPAT3 expression by modulating the AMPK-STAT3 axis. Our study highlights TKFC as a novel and promising target for MASLD and offers new insights into the molecular mechanisms through which Cur ameliorates MASLD.

Recycling of internalized cell surface receptors is critical for membrane transport and receptor-mediated signaling. Formyl peptide receptor 1 (FPR1) plays important roles in host defense and inflammatory tissue injury. Here we report that fMet-Leu-Phe-Cys (fMLFC), a peptide agonist of FPR1, prevents recycling of internalized FPR1 and diverts it to the late endosome and lysosome for degradation. In contrast, FPR1 bound to the classic ligand fMLF interacts with RAB11 and SNX17, facilitating its recycling back to the cell surface. We determined a cryo-EM structure of fMLFC-bound FPR1-Gi complex. Alanine substitutions of key residues that interact with fMLFC (F102A, T177A, F178A) improved FPR1 recycling. Using a FlAsH-NanoBRET-based FPR1 biosensor, the fMLFC-induced receptor conformational change was found to be different from the fMLF-induced conformational change. fMLFC stimulation reduced FPR1 cell surface expression, along with reduced acute lung injury in LPS-treated mice. Our findings suggest that fMLFC is a chemical knockdown agent that promotes targeted protein degradation and reduces FPR1-mediated inflammation.

Hepatic sinusoidal obstruction syndrome (HSOS), a life-threatening liver disease characterized by sinusoidal endothelial cell (LSEC) damage, is frequently caused by pyrrolizidine alkaloid (PA) exposure present in numerous herb or food products. Unlike other hepatotoxins, the precise mechanism by which PAs selectively target LSECs remains poorly understood, posing significant challenges to the development of effective treatments. This study identified hemolysis as the initiating event in PA-HSOS pathogenesis through clinical and animal model analyses. PA exposure induced red blood cell (RBC) rupture, releasing free hemoglobin (Hb) that directly damaged LSECs. Mechanistic investigations revealed that PA-formed protein adducts with haptoglobin (Hp), impairing its protective effect against toxic Hb and triggering a cascade of LSEC activation, ferroptosis, and hemorrhagic liver necrosis. Rescue study revealed that Hp supplementation effectively mitigated PA-induced liver injury by scavenging free Hb. Clinical validation demonstrated elevated Hb-Hp adducts and cell-free Hb in PA-HSOS patients, confirming concordant intoxication mechanisms across species. The findings redefine PA-HSOS as a hematogenous liver disorder originating from RBC destabilization, rather than direct hepatocyte toxicity. This hematopathological perspective reveals Hp replacement therapy as a promising etiological treatment strategy, addressing the root cause rather than secondary liver damage.

Cholangiocarcinoma (CCA) is a markedly desmoplastic tumor, with extrahepatic CCA (eCCA) being the most prevalent subtype. The nonspecific clinical manifestation and early metastatic potential result in a persistently poor overall prognosis. Cancer-associated fibroblasts (CAFs) are one of the major components of the tumor microenvironment implicated in tumor progression and treatment resistance. To faithfully recapitulate the eCCA microenvironment, we established a comprehensive patient-derived preclinical platform comprising CAFs, eCCA primary cells, organoids, and patient-derived xenograft (PDX) models. We found that CAFs facilitated eCCA cell growth, migration, and invasion in vivo and in vitro. Cytokine profiling revealed that HGF is a key paracrine factor produced by CAFs. CAF-derived HGF activates c-MET signaling to drive gemcitabine resistance in eCCA. Therapeutically, pharmacologic inhibition of c-MET partially resensitized eCCA cells to gemcitabine in preclinical models and suppressed tumor progression. In conclusion, our data provides subtype-specific translational evidence that therapeutically targeting the CAF-HGF-c-MET pathway represents a rational strategy to enhance gemcitabine efficacy and improve treatment outcomes in eCCA.

Glioblastoma (GBM), the most aggressive primary brain malignancy, presents an urgent need for novel therapeutic targets addressing metabolic reprogramming in tumor progression. Natural product-based molecular probes have emerged as powerful tools for target discovery and mechanistic elucidation in cancer biology. Here, we identified the fungal polyketide auxarconjugatin B (AUX-B) as a potent inhibitor of GBM proliferation through both in vitro and in vivo models. Chemical biology strategies revealed secretory carrier membrane protein 2 (SCAMP2) as the covalent cellular target of AUX-B. SCAMP2 exhibited significant overexpression in human GBM specimens and orthotopic GBM mouse models, correlating with tumor progression. Mechanistic investigations demonstrated that SCAMP2 orchestrates metabolic reprogramming through the regulation of aspartate transporters (solute carrier family 1 member 3 and solute carrier family 25 member 12) and asparagine synthetase, thereby sustaining aspartate metabolic flux critical for GBM growth. AUX-B-mediated reduction of SCAMP2 effectively disrupted this pathogenic metabolic network, leading to a decrease in intracellular aspartate levels. Our findings establish SCAMP2 as a novel therapeutic target in GBM and characterize AUX-B as a new SCAMP2 inhibitor with translational potential through metabolic modulation.

PACT (PKR activating protein)/PRKRA is a quintessential double-stranded RNA (dsRNA) binding protein that has recently surfaced as a novel and compelling antiviral target, exhibiting resistance against a spectrum of respiratory viruses. Despite this, no antiviral ligand compounds targeting PACT have been identified to date. In this study, we conducted an extensive screening within natural products, leading to the development of an exceptional PACT-T78 site ligand, tubeimoside II (TBM II). TBM II effectively combats a spectrum of respiratory viruses, including the coronaviruses HCoV-OC43 and SARS-CoV-2, as well as the influenza A H1N1 virus (IAV-H1N1), by reducing viral loads and inhibiting viral replication and proliferation both in vitro and in vivo. Single-cell RNA sequencing demonstrated that TBM II significantly impacts the RIG-I signaling pathway associated with PACT. We found that when PACT was knocked down or when RIG-I, MAVS, or IFN-β were knocked out, the ability of TBM II to activate the RIG-I signaling pathway was diminished, resulting in a corresponding attenuation of its antiviral efficacy. These findings indicated that TBM II targets PACT to activate the RIG-I signaling pathway, thereby increasing the secretion of type I interferon IFN-β, ultimately promoting the innate immune response and achieving antiviral efficacy. In summary, our work identified TBM II as a new generation PACT ligand that activating the RIG-I signaling pathway, achieving broad-spectrum antiviral effects.

Triple-negative breast cancer (TNBC) is one of the most aggressive and metastatic forms of breast cancer, for which there are currently no satisfactory therapeutic agents. Here, we reported for the first time that boholamide A, a naturally occurring macrocyclic depsipeptide, exhibited hypoxia-selective anti-TNBC activity against in MDA-MB-231 cells. However, its structure-activity relationships and target had not yet been elucidated. A series of boholamide A analogues were chemically synthesized and evaluated for anti-TNBC potency. The most promising compound 1j was prepared in 13 linear steps with an overall yield of 7.92%, and exhibited high potency against MDA-MB-231 cells with an IC₅₀ value of 0.15 μmol/L under hypoxic condition. Moreover, 1j significantly inhibited proliferation and migration, and induced apoptosis in MDA-MB-231 cells. Compound 16, a prodrug of 1j, significantly inhibited the tumor volume and tumor weight in xenografts. Furthermore, we identified that 1j covalently targeted eukaryotic translation elongation factor 1alpha 1 (eEF1A1) which might underlie the anticancer activity and hypoxia selectivity of boholamide A analogues. These results suggested that boholamide A analogues represented a promising scaffold for discovering hypoxia-selective anti-TNBC agents, and compound 16 deserved further investigation as a candidate for TNBC treatment.

Influenza is a global health issue. Vaccines can protect humans from infection from influenza virus. When no suitable vaccine is available, anti-influenza drugs are the first treatment choice. Emergence of drug resistance necessitates development of novel anti-influenza virus drugs. We investigated a novel macrocyclic compound H1N1-17 using in vitro assays. Through evaluation of its antiviral effect against H1N1 and H3N2 viruses, compound H1N1-17 showed selective inhibition of H1N1 virus strains. Invasion of pseudo-H1N1 was blocked by H1N1-17 in the entry inhibition assay. Membrane fusion of H1N1 and the endosome mediated by the stalk domain of hemagglutinin was inhibited by H1N1-17. In the induction of a drug-resistant mutations assay, the resistant sites of H1N1 to H1N1-17 were located in the F subdomain of hemagglutinin, which is crucial for membrane fusion. Intraperitoneal administration of compound H1N1-17 protected mice challenged with lethal H1N1 from death and weight loss, and effectively alleviated lung injury caused by viral infection. Additionally, compound H1N1-17 exhibited synergistic anti-influenza activity with oseltamivir acid, which was of significance for combination therapy.

The dynamic immune landscape within the tuberculous (TB) granuloma microenvironment critically governs antibiotic penetration efficiency, bacterial persistence, and long-term therapeutic outcomes. Herein, we present a macrophage-targeted inhalable nanoemulsion for co-delivering rifampicin and LCL161, an inhibitor of apoptosis protein antagonist. Inhaled mannose-nanoemulsions (named RL-NE@Man) enable granuloma-targeted delivery in TB mice model, increasing infected macrophage apoptosis, remodeling the tuberculosis microenvironment, and promoting T-cell immunity to synergize with antibiotics for the eradication of granulomas and persistent Mycobacterium tuberculosis infection. Following two-dose inhalational administration, RL-NE@Man displayed potent bactericidal activity against M. tuberculosis while concurrently alleviating pulmonary pathological lesions and hyperinflammatory responses, demonstrating superior bacterial suppression efficacy compared with that of the first-line rifampicin monotherapy. This inhaled combination therapy, which integrates immunomodulators with antibiotics to modulate the local immune landscape and synergistically enhance bactericidal efficacy, represents a novel therapeutic strategy for precision tuberculosis management.

Systemic administration for the clinical management of inflammatory bowel disease (IBD) often leads to various side effects and toxicities, primarily due to broad therapeutic exposure of non-target tissues. Herein, a smart single-atom nanozyme (SAzymes) delivery system (Fe-SA/Cur@HAD, FCH) is constructed through coordinating iron-doped SAzymes (Fe-SA) with curcumin (Cur) for IBD synergistic therapy. Inspired by Trojan horse, FCH efficiently responds to the IBD pathological microenvironment and realizes targeted delivery via oral administration. In inflamed colonic tissue, FCH regulates redox homeostasis through the superoxide dismutase (SOD)-catalase (CAT) cascade reaction and releases Cur by changing the adsorption energy, thus achieving synergistic therapy. An in vitro IBD model of human-derived colonic organoid, along with in vivo IBD model of mouse, were used and demonstrated that this system could effectively reduce reactive oxygen species (ROS) levels, improve intestinal homeostasis, and promote tissue recovery. Additionally, FCH markedly suppresses the activation of inflammatory pathways and modulates the composition of the intestinal microbiota. This study innovatively modifies SAzymes, offering new perspectives on their potential applications in IBD treatment.

Melanoma, the most aggressive form of skin cancer, remains a formidable therapeutic challenge. While oncolytic viruses (OVs) exhibit promising antitumor potential, their efficacy is often limited by insufficient intratumoral viral replication, poor tissue penetration, and the immunosuppressive tumor microenvironment (TME). Herein, a multistage microneedle (MN-OJ) system designed to amplify both local oncolysis and systemic antitumor immunity mediated by oncolytic adenovirus (OA) in melanoma. The dissolvable MN base facilitates rapid OA delivery, inducing tumor cell lysis and subsequent release of tumor-associated antigens to prime T-cell responses. Concurrently, the degradable MN tip enables sustained release of JQ1, which enhances OA replication, modulates lactic acid levels and PD-L1 expression, thereby reprogramming the immunosuppressive TME to promote T-cell infiltration and cytotoxicity. In murine models, MN-OJ demonstrated potent inhibition of both primary and distal tumors, without systemic toxicity. This innovative platform combines immediate tumor destruction with sustained immune modulation, offering a promising clinical approach for melanoma therapy.

The efficient targeted delivery and on-demand release of nanomedicines still present significant challenges in tumor therapy. In the present study, we developed a novel strategy of tumor ferroptosis therapy through the synergistic effect of nanomedicines (biomimetic liposomes) and a medical device (high-intensity focused ultrasound, HIFU). It was found that HIFU irradiation induced heightened expressions of CD44 and reactive oxygen species (ROS) in tumor cells by 1.43-fold and 2.64-fold, respectively. This allowed the gambogic acid-loaded, platelet-mimicking liposomes (PLip) to more precisely target the tumor cells through the interaction of CD44 and P-selectin on the PLip and subsequently ROS-responsively release the drug from PLip, thus effectively killing tumor cells. Crucially, gambogic acid also significantly enhanced ROS production, leading to lipid peroxidation and augmented ferroptosis induced by HIFU. In summary, HIFU demonstrates immense potential in synergizing with nanomedicines to combat tumors.