Acute pancreatitis(AP) is a severe inflammatory disease associated with high morbidity and mortality. Emerging evidence suggests that neutrophil extracellular traps (NETs) play a pivotal role in AP pathogenesis; however, effective therapeutic targets and interventions remain poorly understood.We conducted weighted gene co-expression network analysis (WGCNA) on two independent transcriptomic datasets from the Gene Expression Omnibus to identify disease-associated gene modules. Differential expression analysis of NET-related genes was performed, followed by functional enrichment analysis and protein-protein interaction network construction to elucidate biological pathways. Core genes were identified using least absolute shrinkage and selection operator regression, random forest, and support vector machine recursive feature elimination algorithms. Network pharmacology was employed to predict potential therapeutic drugs. Finally, experimental validation was conducted using a caerulein-induced AP mouse and cell model.The WGCNA analysis identified 2707 key genes. Subsequently, combining differential expression analysis with machine learning approaches, we identified three core genes: glutathione S-transferase mu 1 (GSTM1), syndecan-1 (SDC1), and vitronectin (VTN). These genes exhibited significant expression alterations in AP. Drug prediction and molecular docking analysis revealed that semaxanib demonstrated strong binding affinity to all three target proteins. Experimental validation in the AP model confirmed the bioinformatics predictions: GSTM1 and VTN were significantly downregulated, while SDC1 was markedly upregulated.We identified NETs-related genes GSTM1, SDC1, and VTN as novel biomarkers for AP. Furthermore, semaxanib may have potential associations with pathways related to GSTM1, SDC1 and VTN, thereby participating to some extent in the intervention of acute pancreatitis. These findings provide a foundation for precision medicine approaches in AP management.
BACKGROUND:Atherosclerosis is a chronic vascular disease characterized by endothelial dysfunction and lipid accumulation. Ferroptosis, an iron-dependent form of regulated cell death characterized by excessive lipid peroxidation, plays a pivotal role in endothelial injury and the progression of atherosclerosis. Fibroblast growth factor 21 (FGF21) has been reported to protect against atherosclerosis; however, its role and underlying mechanisms in hypercholesterolemia-induced endothelial ferroptosis remain unclear. METHODS:FGF21 expression and ferroptosis were assessed in vascular endothelial cells from high-fat diet (HFD)-fed ApoE-/- mice and oxidized low-density lipoprotein (ox-LDL)-treated human umbilical vein endothelial cells (HUVECs). Endothelial dysfunction and ferroptosis were evaluated using western blotting, immunofluorescence staining, transmission electron microscopy, mitochondrial membrane potential analysis, and immunohistochemistry. RESULTS:FGF21 expression and ferroptosis were markedly increased in vascular endothelial cells from HFD-fed ApoE-/- mice and ox-LDL-treated HUVECs. Recombinant FGF21 significantly alleviated ox-LDL-induced endothelial dysfunction by inhibiting ferroptosis, with effects similar to those of the ferroptosis inhibitor ferrostatin-1. Mechanistically, FGF21 suppressed endothelial ferroptosis by activating the AMPK/Nrf2 signaling pathway. In vivo, FGF21 treatment markedly reduced atherosclerotic lesion formation and endothelial ferroptosis in HFD-fed ApoE-/-mice, accompanied by enhanced AMPK/Nrf2 signaling activity. CONCLUSION:FGF21 attenuates atherosclerotic progression, at least partially, by alleviating endothelial ferroptosis. AMPK/Nrf2 signaling pathway contributes to the protective effects of FGF21 against atherosclerosis.
BACKGROUND:Diabetic wound healing represents a critical global health challenge, with India harbouring over 77 million diabetic adults and diabetic foot ulcer prevalence ranging from 4.5 to 15%. The complex pathophysiology involves chronic inflammation, impaired angiogenesis, and excessive oxidative stress that fundamentally disrupts normal healing cascades. Luteolin, a naturally occurring flavonoid with documented anti-inflammatory and antioxidant properties, has emerged as a promising therapeutic candidate. OBJECTIVE:This study elucidated the molecular mechanisms of luteolin in diabetic wound healing through an integrated computational and experimental approach. METHODS:Transcriptomic analysis was conducted on three diabetic wound datasets (GSE80178, GSE134431, GSE199939) using differential gene expression analysis. Swiss Target Prediction identified luteolin targets, followed by the construction of a protein-protein interaction network and molecular docking with 200 ns molecular dynamics simulations. Experimental validation involved HS27 human dermal fibroblasts with MTT viability assays, flow cytometry-based ROS measurement, apoptosis analysis, LC-MS/MS metabolomics profiling, scratch wound healing assays, Western blotting, and immunofluorescence. RESULTS:Analysis identified 520 common differentially expressed genes among three datasets and PIK3R1 as a candidate therapeutic target through bioinformatics, machine learning and target prediction. Molecular docking revealed strong binding affinities: PIK3R1 (-9.62 kcal/mol), mTOR (-9.10 kcal/mol), and AKT1 (-8.42 kcal/mol). Treatment with luteolin (25 μg/mL) significantly enhanced cell viability to 130%, markedly reduced intracellular ROS levels (p < 0.001) and promoted wound closure by nearly 40%. Western blot analysis further confirmed the molecular basis of these effects, demonstrating pronounced upregulation of p-PI3K, AKT, NRF2 and Bcl-2 along with concomitant downregulation of GSK-3β and Bax. Metabolomic profiling identified luteolin-responsive metabolites with significant alterations in specific metabolic pathways, including L-leucine and sphingomyelin-related metabolites, were of particular interest given their established association with PI3K/AKT signalling. CONCLUSION:Our study provides a hypothesis linking luteolin to PIK3R1 in diabetic wound-associated dermal fibroblasts, supported by computational, cellular, and metabolomic evidence, and suggests the potential of luteolin as a therapeutic approach for diabetic wound healing.
Tissue-resident memory T (Trm) cells are a specialized subset of memory T cells that persist in peripheral tissues (skin, joints, intestine) without recirculating, serving as a frontline defense against pathogen reinfection. This protective function is exemplified by interleukin-15 (IL-15), C-X-C chemokine receptor type 6, and inducible T-cell co-stimulator signaling that are essential for Trm-mediated immunity against viruses and bacteria. However, the same long-lived persistence becomes detrimental in autoimmunity. Activated autoreactive Trm cells accumulate in lesions of psoriasis, rheumatoid arthritis, inflammatory bowel disease, vitiligo, and other conditions. Upon activation by autoantigens or inflammatory cues, they settle in tissues, continuously produce pro-inflammatory cytokines (interferon-gamma, tumor necrosis factor-alpha, IL-17), directly mediate tissue damage, and recruit other immune cells, thereby amplifying chronic inflammation. Their persistence drives disease relapse, lesion chronicity, and tissue-specific confinement. Thus, Trm cells embody a double-edged sword-essential for local immune protection but also important connectors linking autoimmune initiation to chronic pathology maintenance. In this review, we summarized recent advances in Trm cell biology and examined their growing implications in the pathogenesis of autoimmune diseases. We further discuss the molecular and cellular mechanisms through which Trm cells drive organ-specific, chronic inflammatory responses, underscoring their role across diverse autoimmune disorders. Hopefully, the emerging therapeutic strategies aiming at modulating Trm cell generation, persistence, and function will highlight both the promise and obstacles in translating these approaches into clinical practice.
Knee osteoarthritis (KOA) is characterized by inflammation, extracellular matrix (ECM) degradation, oxidative stress, and chondrocyte loss. Increasing evidence indicates that inflammation-driven redox imbalance and ferroptotic cell death jointly contribute to cartilage degeneration, yet effective pharmacological strategies capable of simultaneously restoring antioxidant defense and suppressing ferroptosis remain limited. This study investigated whether chlorogenic acid (CGA) protects against KOA by regulating Nrf2-associated antioxidant defense and ferroptotic injury. In IL-1β-stimulated ATDC5 chondrocytes, CGA reduced inflammatory mediator expression, restored Aggrecan and Collagen II, and suppressed MMP3 and MMP13. CGA also decreased reactive oxygen species (ROS), lipid peroxidation, mitochondrial dysfunction, and intracellular Fe2+ accumulation while restoring SLC7A11, GPX4, and FTH1 expression. In an RSL3-induced ferroptosis model, both CGA and ferrostatin-1 attenuated Fe2+ accumulation, malondialdehyde (MDA) production, ferroptosis-defense impairment, and loss of glutathione peroxidase (GPx) activity. CGA enhanced nuclear Nrf2 accumulation and downstream HO-1 signaling, whereas Nrf2 silencing substantially attenuated its antioxidant, matrix-preserving, and anti-ferroptotic effects. In the destabilization of the medial meniscus-induced KOA mouse model, CGA alleviated cartilage degeneration, synovial inflammation, ECM disruption, and ferroptosis-associated changes without overt organ toxicity. These findings extend the established anti-inflammatory and antioxidant actions of CGA by providing functional evidence that CGA attenuates ferroptotic damage and that Nrf2 contributes importantly to its chondroprotective effects.
Glutaminyl-peptide cyclotransferase-like protein (QPCTL) is a pivotal post-translational modification enzyme remodeling tumor microenvironment through catalyzing the pyroglutamation (pGlu) at the N-terminus of CD47, CCL2 and CCL7. Inhibiting QPCTL attenuates the CD47-SIRPα interaction and impairs the recruitment of pro-tumoral macrophages, thereby promoting anti-tumor immunity, and has emerged as a potential target for cancer immunotherapy. Nevertheless, the development of QPCTL inhibitors for cancer immunotherapy is still in its early stages. Herein, we report the discovery and characterization of QP6126, a highly potent and orally bioavailable small-molecule QPCTL inhibitor (IC50 of 2.3 nM). Mechanistically, QP6126 effectively ablates the pGlu-CD47 modification, thereby abolishing CD47-SIRPα interaction, and sensitizing melanoma cells to macrophage-mediated phagocytosis. Notably, QP6126 displays significant in vivo antitumor efficacy in B16F10 melanoma mouse model following oral administration. This study establishes QP6126 as a promising candidate for clinical translation and validate QPCTL as a strategic target for diversifying cancer immunotherapy.
PURPOSE:To investigate the role and mechanisms of calcitriol in form-deprivation myopia (FDM). METHODS:Calcitriol was evaluated in a guinea pig FDM model and a tumor necrosis factor-alpha (TNF-α)-induced inflammatory model using human retinal pigment epithelial (RPE) cells. Vitamin D receptor (VDR) dependence was assessed by adeno-associated virus- or small interfering RNA-mediated VDR knockdown. Refractive status, axial length, fundus changes, scleral collagen remodeling, inflammatory cytokines, nuclear factor kappa-B (NF-κB) activation, and matrix metalloproteinase-2 (MMP2) expression were examined. Protein interaction analysis explored VDR-NF-κB p65 interaction. RESULTS:Calcitriol significantly attenuated FDM progression in guinea pigs, reducing myopic refractive shift, axial elongation, and fundus alterations. It increased scleral collagen expression and improved collagen organization and ultrastructure. Calcitriol also decreased circulating and retinal TNF-α, interleukin-1β, and interleukin-6 levels, accompanied by NF-κB inhibition and MMP2 downregulation. VDR knockdown markedly weakened the protective effects of calcitriol on refractive and axial changes, inflammatory cytokine production, and NF-κB/MMP2 signaling. In vitro, calcitriol inhibited TNF-α-induced NF-κB p65 nuclear translocation and reduced inflammatory cytokine secretion and MMP2 expression in RPE cells; these effects were largely abolished by VDR knockdown. Protein interaction analysis suggested that calcitriol enhanced VDR-NF-κB p65 interaction, thereby restricting NF-κB activation. CONCLUSIONS:Calcitriol delays FDM progression through VDR-mediated inhibition of NF-κB activation, reducing inflammatory cytokine production, MMP2 expression, scleral collagen degradation, and tissue remodeling.
The intent of the current study was the repurposing of rosuvastatin (RSV), an anti-hyperlipidemic drug, for the management of rheumatoid arthritis (RA). Transdermal delivery of RSV, a repurposed drug, loaded in nanoemulgel (NEG) improves the permeation of the drug through the skin and reaches the desired site of action. This thereby enhances the concentration of the drug at the site of action, reducing the side effects associated with other routes of delivery, further showing anti-inflammatory potential in the management of RA. Nanoemulsion (NE) was prepared by the spontaneous emulsification method. The globule size and polydispersity index of the developed NE were evaluated by Malvern ZetaSizer. Furthermore, RSV-NEG has been developed and characterized. In vivo efficacy of RSV-NEG was evaluated in complete Freund's adjuvant-induced RA in rats. Pro-inflammatory cytokine concentrations in rat serum were quantified by enzyme-linked immunosorbent assay. The globule size and polydispersity index of 0.4% w/w RSV NE were found to be 8.96 ± 0.19 nm and < 0.3, respectively. The developed NEG showed pseudoplastic shear thinning behavior and an average drug content of 97.54 ± 0.36%. The RSV demonstrated 28-fold higher permeation from RSV-NEG compared to free RSV gel. The animals treated with RSV-NEG exhibited anti-inflammatory potential along with significantly reduced (****p < 0.0001) paw thickness and arthritis scores compared to the negative control group. The promising findings with permeability after topical application and anti-inflammatory potential emphasize the ability of RSV-NEG to manage RA effectively.
BACKGROUND:Intrauterine adhesion (IUA) significantly contributes to female infertility, primarily characterized by abnormal fibrotic remodeling of the endometrium. The intricate interplay among various cellular components within the endometrial microenvironment plays a pivotal role in IUA pathophysiology. Among these implicated factors, the vitamin D receptor (VDR) has emerged as a potential regulator of fibrotic processes; however, its specific role and underlying mechanisms in IUA remain inadequately understood. This study aimed to investigate the role of VDR in IUA pathogenesis and its impact on endometrial macrophage behavior. METHODS:To investigate VDR's role in IUA, we conducted a comprehensive analysis of clinical endometrial samples from patients diagnosed with IUA. Complementary to this, we performed in vitro experiments using bone marrow-derived macrophages (BMDMs) obtained from both wild-type (WT) and VDR conditionnal knockout (VDR-cKO) mice. Furthermore, in vivo studies were carried out using an IUA mouse model alongside VDR-cKO mice to assess the physiological relevance of our findings. RESULTS:Our investigation revealed a marked reduction in VDR expression within endometrial macrophages from IUA patients. In vitro assessments indicated that VDR deficiency led to enhanced pro-inflammatory M1 polarization, evidenced by increased level of iNOS and decreased CD206 in BMDMs. Mechanistically, VDR conditional knockout was found to induce cuproptosis, characterized by downregulation of ATP7B and upregulation of DLAT. Additionally, we demonstrated that VDR transcriptionally activates ATP7B expression, with luciferase reporter assays confirming VDR's direct binding to the ATP7B promoter. In vivo experiments further established that macrophage-specific VDR knockout intensified endometrial fibrosis, heightened M1 polarization, and increased cuproptosis in the IUA mouse model, while administration of a VDR agonist effectively reversed these phenotypic alterations. CONCLUSION:Our findings reveal a novel mechanistic pathway by which macrophage-specific VDR deficiency exacerbates IUA pathogenesis through dysregulation of the ATP7B-cuproptosis axis and promotion of profibrotic M1-like macrophage polarization. These insights suggest that VDR agonism may represent a promising therapeutic approach for the prevention and treatment of IUA, warranting further investigation into its clinical applicability.
Emamectin benzoate (EMB) is a highly effective and widely detected environmental insecticide whose potential to induce severe cardiotoxicity and localized myocardial inflammation has raised substantial biomedical concerns. Quercetin (Que), a ubiquitous plant-derived natural antioxidant, has shown promise in mitigating exogenous chemical-induced toxicities, yet its precise immunopharmacological mechanism against EMB-induced cardiotoxicity remains to be fully elucidated. This study investigated the protective effects and mechanisms of Que. against EMB-induced cardiotoxicity by integrating network toxicology, transcriptomics, and in vivo/in vitro experimental validation. Multi-omics analysis identified MAPK8, MAPK3, and CASP3 as key responsive targets conserved across species. Functional enrichment revealed that these targets are predominantly involved in the MAPK/NF-κB signaling pathway and mitochondrial organization. Molecular docking and targeted intervention experiments confirmed that Que. potentially targets and binds to JNK (encoded by MAPK8), thereby blocking the EMB-activated signaling axis. This molecular interaction effectively restored mitochondrial dynamics and prevented mitochondrial DNA leakage into the cytoplasm, ultimately alleviating myocardial PANoptosis. Collectively, our findings demonstrate that Que. serves as a natural antagonist against chemical-induced cardiotoxicity by modulating the MAPK/NF-κB axis, offering a promising therapeutic strategy for mitigating pesticide-related immunotoxicological risks.
Asthma remains a significant global health burden characterised by high clinical heterogeneity and immunological complexity. Despite currently available therapies, limitations like steroid insensitivity and adverse side effects persist. Furthermore, recent biologics' high costs and accessibility remain a constraint. This emphasised the need for innovative alternative treatments. Kaempferol, a naturally occurring flavanol found in various fruits, vegetables, and medicinal plants, has gained considerable attention for its multifaceted pharmacological properties. This review critically assesses existing preclinical evidence regarding the therapeutic potential and molecular mechanisms of kaempferol in experimental asthma models. Preclinical in vivo and in vitro studies consistently demonstrate that kaempferol targets three critical dimensions of asthma pathogenesis: airway inflammation, hyperresponsiveness, and airway remodelling. Mechanistically, it suppresses the infiltration of inflammatory cells and reduces the secretion of pro-inflammatory cytokines, chemokines, and IgE. These effects are achieved by modulating key signalling pathways, including NF-κB, MAPKs, JAK/STAT, Nrf2, and NOX4-mediated autophagy. Furthermore, kaempferol alleviates smooth muscle hypercontractility and suppresses airway remodelling by mitigating subepithelial fibrosis, mucus hypersecretion, and epithelial-to-mesenchymal transition. Its translational strengths include potential steroid-sparing effects. Despite this, its clinical application is hindered by poor oral bioavailability and a lack of direct human trials. Rigorous clinical validation and advanced targeted delivery systems are required to establish kaempferol as a viable therapeutic agent for asthma management.
This correspondence provides a critical perspective on the recent study by Zhao et al. defining the hyperglycemia-STAT3-neutrophil extracellular trap-thromboinflammation axis in post-ischemic stroke hemorrhagic transformation. Beyond three core unresolved scientific gaps concerning cell-type specificity, tissue plasminogen activator-associated pathological context, and glycemic model heterogeneity, we extend the discussion to supplementary translational constraints, the upstream regulatory role of mitochondrial oxidative stress, and implications for clinical trial design. These viewpoints aim to further enhance the translational potential of this promising therapeutic target.
Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) is a common urological condition marked by ongoing oxidative stress and persistent inflammation, which complicates clinical management. This study examined the therapeutic effects and underlying mechanisms of 7,8-Dihydroxyflavone (7,8-DHF) in an experimental autoimmune prostatitis (EAP) mouse model, induced via subcutaneous immunization with prostate antigen in Complete Freund's Adjuvant. Mice were administered daily intraperitoneal injections of 7,8-DHF (5 mg/kg) and subsequently underwent a series of evaluations: prostate index measurement, abdominal tactile allodynia assessment using Von Frey filaments, cystometric analysis for urinary function, histological examinations with HE and Masson staining, quantitative PCR for pro-inflammatory cytokines, biochemical assays for oxidative stress markers (MDA, SOD, CAT, total antioxidant capacity), and Western blot analysis to assess activation of the TrkB/AKT/ERK/SIRT3 signaling pathway. 7,8-DHF treatment significantly decreased the prostate index, alleviated pelvic tactile allodynia, and improved bladder function in EAP mice. Histopathological analysis showed notable reductions in inflammatory cell infiltration and collagen deposition, along with decreased expression of pro-inflammatory cytokines (IL-6, IL-17α, MCP-1, IL-1β). Mechanistically, 7,8-DHF enhanced antioxidant capacity by increasing SOD/CAT activities and decreasing MDA levels, which correlated with TrkB phosphorylation, subsequent AKT/ERK pathway activation, upregulated SIRT3 expression, and MnSOD deacetylation. In vitro experiments with RWPE-1 cells demonstrated that 7,8-DHF mitigates LPS-induced oxidative stress and inflammation through mechanisms dependent on SIRT3. Collectively, these findings elucidated that 7,8-DHF could ameliorate CP/CPPS by activating the TrkB/AKT/ERK/SIRT3 signaling pathway to suppress oxidative stress and inflammatory reactions, highlighting its potential value as a novel therapeutic candidate for the clinical management of CP/CPPS.
BACKGROUND:Renal aging accelerates overall organismal aging. The traditional Chinese medicine formula Huyan I (HY-1) has been shown to slow the decline in kidney function, but its pharmacological mechanism is unclear. This study focuses on its active component, madecassoside (MA), aiming to elucidate how it delays renal aging. METHODS:HY-1 was evaluated in a D-galactose-induced mouse model using renal functional, histopathological, inflammatory, and molecular assessments. Serum-absorbed constituents were characterized by UHPLC-MS/MS and analyzed using network pharmacology, molecular docking, MM/GBSA binding free-energy calculations, and molecular dynamics simulations. The anti-senescent effects of madecassoside (MA) were validated in vivo and in D-galactose-treated HK-2 cells. Transcriptomic sequencing and pharmacological pathway modulation were used to investigate the involvement and hierarchical relationship of the AMPK and MAPK/AP-1 signaling axes. RESULTS:Both HY-1 and MA ameliorated renal pathological injury, improved the expression levels of aging-related proteins (p53, p21, p16), promoted AMPK phosphorylation, and reduced senescence-associated secretory phenotype (SASP) factors (IL-1β, IL-6, TNF-α) in mouse serum, thereby slowing overall aging. MA also reduced intracellular peroxide accumulation in HK-2 cells, restored mitochondrial membrane potential, and improved mitochondrial function. Transcriptomic analysis revealed that MA may exert anti-aging effects by suppressing the MAPK signaling pathway (inhibiting phosphorylation of p38, ERK, JNK) and reducing nuclear phosphorylation of c-Fos and c-Jun, affecting AP-1 transcription factor formation. When AMPK phosphorylation was inhibited, MA's anti-aging effect was abrogated, and MAPK pathway activation was enhanced. CONCLUSION:As the one of the principal active ingredients of HY-1, MA delays renal aging by promoting AMPK phosphorylation to maintain mitochondrial function and reduce peroxide accumulation, while inhibiting the MAPK signaling pathway to decrease AP-1 transcription factor synthesis.
Proper immune response needs a regulated interaction between adaptive and innate immunity and dendritic cells (DCs) as professional antigen presenting cells coordinate their interaction. In order to maintain the balance between tolerance and initiation of immune response, these DCs should be activated in a regulated manner which is managed by diverse signaling pathways including nuclear factor kappa light chain enhancer of activated B cells (NF-κβ) and NLR family pyrin domain containing 3 (NLRP3). According to the vital role of DCs in immunity, it is a promising strategy to induce tolerogenic DCs (TolDCs) to suppress autologous T cell responses in patients with autoimmune disorders and targeting inflammatory signaling pathways is a potent strategy for induction of mentioned TolDCs. NF-κβ and NLRP3 signaling pathways play significant role in maturation, antigen presentation, and inflammatory function of DCs and finally affect their immune response which subsequently contributes to differentiation of T cells. According to the stated vital role of signaling pathways in DCs and their involvement in progression of autoimmune disorders, in this study we will review NF-κβ and NLRP3 signaling pathways and their contribution to DCs' immunogenicity and functionality and it will be discussed that how these signaling pathways are strictly regulated. Furthermore, this review sheds light on therapeutic role of NF-κβ and NLRP3 targeting for induction of potent TolDCs for the benefit of patients with diverse autoimmune disorders. Finally, due to the well-known role of NF-κβ as primary needed signal for assembly and full function of NLRP3 inflammasome we will mention the cross-talk between NF-κβ and NLRP3 pathways and highlight their interrelated targeting as a mechanistically rational approach that warrants future experimental investigation to evaluate effects of their simultaneous modulation for induction of favorable TolDC vaccines with both NF-κβ and NLRP3 being suppressed that not only disrupts the immunogenic function of DCs by inhibition of NF-κβ, but also prevents full activation of NLRP3 inflammasome.
Interleukin-15 (IL-15) is a potent immunostimulatory cytokine but is limited by rapid clearance and systemic toxicity. To overcome these limitations, our laboratory generated a chimeric IL-15 by fusing human IL-15 to the murine IgG2a Fc domain and evaluated its activity, efficacy, pharmacokinetics, and safety. The purified protein showed accurate molecular integrity determined through SDS-PAGE and Western blot. In-vitro, chimeric IL-15 induced dose-dependent activation of CD4+ and CD8+ T cells and NK cells, with increased frequencies of CD8+CD107a+ cytotoxic T cells and CD8+IFN-γ+ effector populations. In-vivo treatment resulted in time- and dose-dependent expansion of splenic cytotoxic lymphocyte subsets. Pharmacokinetic analysis demonstrated rapid distribution and a prolonged half-life of 40-53 h, while biodistribution studies confirmed widespread tissue localisation without abnormal accumulation. In syngeneic B16F10 melanoma and 4T1 breast cancer models, chimeric IL-15 significantly inhibited tumour growth and improved survival, accompanied by increased infiltration and activation of CD8+ T cells and NK cells in tumours and lymphoid tissues. Acute and subacute toxicity studies revealed no major haematological, biochemical, or histopathological abnormalities, with only transient, reversible elevations in liver enzymes at higher doses. Collectively, these findings demonstrate that chimeric IL-15 induces robust antitumor immunity with improved pharmacokinetics and a favourable safety profile, supporting its further preclinical development as a next-generation cancer immunotherapeutic.
SIGNIFICANCE:In the case of heterogeneous autoimmune diseases (AIDs), the efficacy of single-pathway immunosuppression is limited, the toxicity is substantial, and the problem of cell type-specific metabolic vulnerability cannot be addressed. MECHANISM:New evidence highlights mitochondrial quality control (MQC) as an important regulatory axis of the immune set point. MQC is a coordinated network involving mitochondrial biogenesis, dynamics, mitophagy, proteostasis, protein import machinery, and selective removal of damaged mitochondrial components, including mitochondria-derived vesicles. Dysregulation across these interconnected MQC modules may contribute to key pathological events: impaired biogenesis compromises the metabolic fitness of regulatory T cells; an altered fission-fusion balance can influence macrophage inflammatory polarization; and defective mitophagy may increase the accumulation or release of immunostimulatory mitochondrial components, including mitochondrial DNA (mtDNA), triggering cyclic GMP-AMP synthase (cGAS)-Stimulator of Interferon Genes (STING)-driven type I interferon (IFN-I) amplification and NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome hyperactivation. In parallel, impaired mitochondrial proteostasis, defective mitochondrial protein import machinery, dysregulated mitochondrial unfolded protein response (UPRmt) signaling, and altered Mitochondrial-derived vesicle (MDV)-mediated cargo disposal may further shape mitochondrial stress and immune-cell function. INNOVATION:We propose a conceptual shift from blanket immunosuppression to potential precision metabolic-immune reprogramming. This framework proposes that accessible MQC-related readouts, including transcription factor A (TFAM), Dynamin-related protein1 (Drp1) phosphorylation patterns, and Microtubule-associated protein 1A/1B-light chain 3 (LC3), may help inform treatment timing and therapeutic windows in future translational studies. FUTURE DIRECTIONS:To advance potential medicine in AIDs, prospective clinical studies should evaluate whether MQC-related biomarkers can identify treatment-responsive subgroups. For example, future studies should assess whether disease-stage and cell-specific modulation of mitophagy or mitochondrial dynamics improves treatment responses in rheumatoid arthritis (RA) patients stratified according to synovial Translocase of Outer Mitochondrial Membrane 7 (TOMM7) expression and metabolic state. These studies should test whether fission inhibition, restoration of regulated fission, or normalization of dysregulated fusion is beneficial in specific patient subgroups.
Sepsis-associated acute respiratory distress syndrome (S-ARDS) is often followed by immunoparalysis, leaving patients vulnerable to secondary infection and adverse outcomes. Lactate is widely used as a severity marker in sepsis, but its direct contribution to immune paralysis remains unclear. We examined lactate-driven immune dysfunction in mouse models, bone marrow-derived macrophages and human CD14+ monocyte-derived macrophages, and tested C646-loaded mesenchymal stromal cell-derived extracellular vesicles (EVs-C646) as an experimental intervention. Lactate elevation blunted cytokine responses after rechallenge, reduced macrophage phagocytosis, shifted macrophages toward an M2-like phenotype, expanded regulatory T cells and decreased interferon-γ (IFN-γ) responses in CD8+ T cells. Mechanistically, lactate activated a p300-histone H3 lysine 18 lactylation (H3K18la) program and upregulated proteasome 26S subunit, non-ATPase 14 (PSMD14). Integrated RNA sequencing, H3K18la chromatin immunoprecipitation sequencing, public sepsis transcriptomic analysis and perturbation experiments identified PSMD14 as a lactylation-linked effector that strengthened AKT/mTOR signalling and promoted macrophage dysfunction. EVs-C646 decreased H3K18la and PSMD14 expression, restored macrophage inflammatory responsiveness, improved bacterial clearance, reduced lung injury and increased survival in experimental S-ARDS-related immunoparalysis models. These findings define a lactate-p300-H3K18la-PSMD14-AKT/mTOR pathway in immunoparalysis and support further preclinical evaluation of lactylation-targeted extracellular vesicle therapy.
Copper (Cu) is an essential mineral nutrient in animal, however, excess Cu causes mitochondrial homeostasis imbalance and multiple regulated cell death. This study explored how Cu overload disrupts the SAM50-MICOS-ATAD3-mtDNA axis and further activates cGAS-STING-NLRP3-mediated pyroptosis in pig liver via in vivo feeding model and in vitro hepatocyte with Cu chelator TTM, SAM50 overexpression and H₂O₂ positive control. We verified that excess dietary Cu triggers hepatic lipid metabolic disorder and impairs mitochondrial respiratory activity. Mechanistically, Cu weakens bidirectional protein interactions within the SAM50-MICOS-ATAD3 complex without altering total protein abundance of these subunits, thus breaking mtDNA retention homeostasis. Therefore, Cu exposure significantly increased the mtDNA (D-LOOP, mtATP6, mtCO1, mtND1, mtCO2 and mtND2) released to cytoplasm and plasma compared with that in the control group and raised the level of IFN-β (P < 0.05). Additionally, Cu exposure significantly increased the mRNA and protein expression of cGAS-STING pathway and NLRP3 dependent pyroptosis compared with the control group, while obviously decreased the mRNA and protein expressions of TFAM. Immunohistochemistry and immunofluorescence validated elevated GSDMD and NLRP3 as well as reduced TFAM in Cu stressed liver tissue. Notably, SAM50 overexpression effectively restored the disrupted complex, curtailed mtDNA egress, and attenuated downstream inflammatory activation, phenocopying the protective effects of TTM chelation. Collectively, our findings demonstrated Cu overload destroyed the stability of SAM50-MICOS-ATAD3-mtDNA axis, promotes mtDNA released and triggered the cGAS-STING and NLRP3 pathways in pig hepatocytes.
Background Asthma is a chronic inflammatory airway disease characterized by Th2- dominant immune responses, airway hyperresponsiveness, and structural remodeling. Although inhaled corticosteroids and biologics are effective for many patients, a substantial proportion remains poorly controlled and experiences treatment-related adverse effects. Probiotics have emerged as immunomodulatory agents in asthma, but existing studies predominantly focus on oral administration and gut-lung axis regulation. Whether direct respiratory administration of probiotics can modulate the pulmonary immune microenvironment and alleviate asthma remains largely unexplored. Methods An ovalbumin (OVA)-induced asthma model was established in C57BL/6 J mice. Clostridium butyricum, Lactobacillus casei, or Bifidobacterium infantis were administered intranasally during the challenge phase. Inflammation and remodeling were evaluated using bronchoalveolar lavage fluid (BALF) cell counts, serum IgE measurement, and histological staining (H&E, PAS, Masson). Lung immune and stromal cell heterogeneity and intercellular interactions were analyzed by single-cell RNA sequencing, while airway microbiota composition was characterized by 16S rDNA sequencing. Results Intranasal probiotic administration attenuated OVA-induced airway hyperresponsiveness, eosinophilic inflammation, and was associated with reduced mucus hypersecretion and collagen-associated histological changes. Probiotics suppressed Th2-biased immune responses, evidenced by reduced Th2 cell proportions, downregulation of Gata3, and decreased expression of Il4, Il5, and Il13. These effects were associated with inhibited dendritic cell activation and weakened DC-T cell interactions, particularly via the Cxcl16-Cxcr6 axis. Probiotic treatment was associated with an increased proportion of M2 macrophages, reduced pro-inflammatory signaling, reduced predicted macrophage-fibroblast communication through the Osm-Osmr pathway, and a lower proportion of fibrotic fibroblasts. Additionally, intranasal probiotic treatment was associated with changes in airway microbial composition, including a reduced relative abundance of Neisseria, which was positively correlated with Th2 cytokine expression. Conclusion Intranasal probiotic administration was associated with attenuation of airway inflammation and early remodeling-associated changes, accompanied by coordinated changes in Th2 immunity, dendritic cell activation, macrophage polarization, fibroblast subtypes, and the airway microbiota. These effects appeared to be strain-specific, with Lactobacillus casei, Bifidobacterium infantis, and Clostridium butyricum being predominantly associated with suppression of Th2 immunity, modulation of predicted DC-T cell communication, and macrophage polarization, respectively. These findings provide preclinical evidence supporting further investigation of intranasal probiotics as a potential strategy for asthma management.
