Aerobic exercise reduces cardiovascular disease risk, with atherosclerosis being a primary contributor. While circulating extracellular vesicles (EVs) mediate intercellular communication, their role in this process remains unclear. This study aimed to investigate the role of aerobic exercise-derived circulating EVs in mitigating macrophage inflammation and lipid accumulation in an atherosclerotic model. Circulating EVs were isolated from the plasma of exercise-trained and sedentary mice. miRNA profiling of EVs was performed using miRNA arrays and quantitative real-time PCR. Aortic atherosclerosis was assessed by Oil Red O staining, immunofluorescence, and ELISA. Functional validation of EV effects was carried out through EVs labeling, cell transfection, luciferase reporter assays, and flow cytometry. Aerobic exercise slowed the progression of atherosclerosis and altered the miRNA profile of circulating EVs, notably increasing miR-203a-3p and miR-133b-3p expression. EVs from exercise-trained mice inhibited macrophage-driven inflammation and lipid accumulation in vitro and in vivo. Treatment with miR-203a-3p and miR-133b-3p mimics reproduced the anti-atherosclerotic effects, while inhibitors of these miRNAs reversed the effects. Mechanistically, miR-203a-3p and miR-133b-3p reduced macrophage inflammation and lipid accumulation by targeting Tlr4 and Insr, respectively, thereby suppressing NF-κB/NLRP3 signaling. Notably, the increased expression of miR-203a-3p and miR-133b-3p was primarily derived from skeletal muscle. These findings highlight a novel mechanism linking aerobic exercise to atherosclerosis via EV-miRNAs, proposing potential therapeutic strategies for atherosclerosis based on exercise-induced circulating EVs-miR-203a-3p and miR-133b-3p.
Epidermal keratinocyte differentiation and cell death are tightly regulated to maintain skin homeostasis, and their disruption leads to severe skin disorders. Proteasome dysfunction has been implicated in proteasome-associated autoinflammatory syndromes, which are characterized by dermal inflammation, as well as in keratosis linearis with ichthyosis congenita and sclerosing keratoderma syndrome caused by mutations in proteasome maturation protein. However, skin manifestations in proteasome-related disorders have been attributed to distinct cellular origins, rendering it unclear whether keratinocyte-intrinsic proteasome dysfunction is a common driver of both epidermal pathology and dermal inflammation. Here, we demonstrate that keratinocyte-specific deletion of the standard proteasome subunit β5 results in neonatal lethality associated with complete loss of the epidermis. Inducible deletion of β5 in basal keratinocytes led to impaired epidermal differentiation, reduced proliferation, and apoptosis, accompanied by accumulation of ubiquitinated proteins without evidence of endoplasmic reticulum stress. Notably, proteasome dysfunction induced epidermal hyperplasia with spongiosis, followed by epidermolysis. Although this genetic alteration was restricted to keratinocytes, pronounced infiltration of neutrophils and macrophages was observed in the dermis, together with elevated expression of inflammatory mediators. These findings demonstrate that proteasome dysfunction in basal keratinocytes disrupts epidermal integrity and triggers secondary dermal inflammation, providing insight into the skin pathology associated with proteasome-related disorders.
The liver serves as a key metabolic and inflammatory organ involved in maintaining homeostasis of different organs and systems. This includes ensuring proper functioning of the central nervous system through a liver-brain axis that has bi-directional communication via signaling mediators and direct neural pathways. Damage to, or disease of, the liver from various etiologies leads to a loss of hepatic function which then impacts cognitive, motor, and other neurological functions. Severity, type and duration of liver dysfunction can impact the brain in different ways via different metabolic and immune challenges. Some of the hepatic factors that impact the brain include ammonia, cytokines, and bile acids. These factors can induce more apparent neurological changes, such as hepatic encephalopathy, while others induce more subtle changes which typically fall outside of the classical characterization of hepatic encephalopathy. The aim of this review is to outline the current knowledge of the liver-brain axis in pathological states. Specific emphasis is placed on how loss of liver function leads to systemic metabolic and inflammatory challenges and their subsequent impacts on hepatic and neural pathology. In addition, specific hepatic pathologies most associated with disruptions of the liver-brain axis will be focused on for more detailed analysis.
Oral squamous cell carcinoma (OSCC) is an aggressive malignancy with poor clinical outcomes driven by metastasis and recurrence. Vascular endothelial growth factor receptor 2 (VEGFR2), a key mediator of vascular endothelial growth factor A (VEGFA)-dependent angiogenesis, has recently been implicated in tumor cell-intrinsic signaling; however, its role in OSCC remains unclear. This study investigated VEGFA-VEGFR2 signaling in OSCC using a 4-nitroquinoline 1-oxide-induced mouse carcinogenesis model and human tissue specimens. Spatial transcriptomic analysis of murine tongues demonstrated enrichment of Kdr (Vegfr2) and Vegfa expression in tumor regions. Immunohistochemical analyses demonstrated up-regulation of VEGFR2, particularly nuclear VEGFR2, during tumor development. Exogenous VEGFA increased VEGFR2 expression and induced its nuclear localization in murine carcinoma cells. In human tongue squamous cell carcinoma, VEGFR2 was localized in the cytoplasm and nucleus, and nuclear VEGFR2 expression was significantly associated with tumor stage, lymph node metastasis, and proliferative activity. Nuclear VEGFR2 expression also correlated with VEGFA expression in epithelial and stromal compartments. Functional analyses showed that VEGFA-VEGFR2 signaling enhanced cell migration and stem-like properties independently of angiogenesis. These findings indicate that nuclear VEGFR2 reflects pathway activation and contributes to tumor aggressiveness, highlighting its potential as a biomarker and therapeutic target.
Ferroptosis is an iron-catalyzed lipid peroxidation-dependent cell death that mediates the development of many diseases, including liver injury. Compelling evidence has suggested a crucial role of mitochondrial reactive oxygen species (mtROS) in the induction of ferroptosis, but the underlying mechanism remains poorly defined. In this study, the impact of mtROS-driven signaling on cellular metabolism, oxidation-reduction state, and ferroptosis vulnerability of the hepatocytes was investigated by using mtROS inducers, including iron overload and pharmacologic inducers. Elevations in mtROS production and lipid peroxidation suppressed glycolysis, fatty acid oxidation, and tricarboxylic acid cycle activity, protecting hepatocytes from ferroptosis. In contrast, mtROS-induced signaling down-regulated genes involved in glutathione biosynthesis, and coenzyme Q10 (CoQ) biosynthesis, including those in the mevalonate pathway, and CoQ8A, a key stabilizer of the CoQ biosynthetic complex. Importantly, silencing CoQ8A expression enhanced, whereas overexpression of CoQ8A reduced, ferroptosis susceptibility of the hepatocytes. Further analysis showed that mtROS-mediated down-regulation of CoQ8A is dependent on farnesoid X receptor and retinoid X receptor. Collectively, these findings suggest that mtROS induces down-regulation of glutathione and CoQ biosynthesis, thereby promoting ferroptotic death in hepatocytes.
Gastric gland mucins contain O-glycans carrying terminal α1,4-linked N-acetylglucosamine (αGlcNAc). A previous study demonstrated that A4gnt-deficient mice lack αGlcNAc, spontaneously develop differentiated-type gastric adenocarcinoma, and exhibit significant upregulation of Il11 transcripts in gastric mucosa. This study assessed mechanisms by which αGlcNAc deficiency induces tumorigenesis, focusing on the IL-11Rα-gp130-JAK2-STAT3 axis. A4gnt-deficient mice exhibited enhanced phosphorylation of JAK2 and STAT3 in pyloric mucosa compared with wild-type mice, with significant increases in JAK2 phosphorylation at 5 weeks and in STAT3 phosphorylation at 5 and 10 weeks. IL-11Rα and gp130 proteins also significantly increased during this period. The thickness of pyloric gland cell layer was significantly decreased in A4gnt and Il11 doubly-deficient (A4gnt-/-;Il11-/-) mice at 10 but not 30 weeks compared to A4gnt-deficient mice, suggesting an IL-11 requirement in early stage of tumorigenesis. Significant reduction of the proportion of p-STAT3-positive nuclei in pyloric mucosa of A4gnt-/-;Il11-/- mice at 10 weeks supports the IL-11-mediated p-STAT3 involvement in this stage. Analysis of gastric cancer AGS-A cells overexpressing αGlcNAc using a Tet-On system revealed significant reduction of IL-11-induced STAT3 phosphorylation compared with αGlcNAc-negative conditions. Moreover, both IL-11Rα and gp130 from wild-type mice and αGlcNAc-expressing AGS-A cells carried αGlcNAc, suggesting that the presence of αGlcNAc on these proteins interferes with receptor-complex formation. Overall, these findings indicate that αGlcNAc protects gastric mucosa from inflammation-induced tumorigenesis by regulating IL-11Rα-gp130-JAK2-STAT3 signaling.
Sjögren disease (SjD) is a chronic systemic autoimmune disorder characterized by immune-mediated injury to exocrine glands and multiple extraglandular organs, affecting millions of individuals worldwide. Despite its clinical burden, SjD remains challenging to diagnose in a timely manner because of the limited availability of objective diagnostic tools and the lack of knowledge in its pathogenic mechanisms, which also hinders the development of targeted therapy. Current therapeutic strategies include systemic immunosuppressive agents and corticosteroids, supplemented by symptomatic management; however, these approaches offer variable efficacy and induce systemic adverse effects. This review summarizes recent advances in the clinical diagnosis of SjD, emphasizing evolving diagnostic criteria and advanced diagnostic tools, including ultrasonographic imaging, biopsies, as well as tear/salivary and hematological biomarkers; and discusses current concepts of SjD pathophysiology, focusing on immune dysregulation, inflammatory signaling, and tissue-specific pathologic changes. Moreover, novel therapeutics targeting pattern recognition receptors and lymphocytic activation were included.
Glaucoma is a neurodegenerative eye disease characterized by progressive loss of retinal ganglion cells (RGCs), resulting in irreversible vision loss. Previous work demonstrated that irbesartan, an angiotensin II type I receptor blocker, promotes RGC survival in rat retinal explants. This study extends these findings to ex vivo mouse explants, in vivo optic nerve crush model, and transcriptomic analysis to identify associated pathways. Retinal explants from C57BL/6 mice were cultured for 4 days with irbesartan (10 μmol/L), vehicle, or angiotensin II (2 μmol/L). RGC survival was assessed by βIII-tubulin and RNA-binding protein with multiple splicing (RBPMS) immunostaining; superoxide and mitochondrial density were measured using dihydroethidium and MitoTracker Red CMXRos. Mice received vehicle or irbesartan (10 μmol/L) eye drops for 7 days after optic nerve crush to assess RGC survival. Irbesartan enhanced RGC survival in explants (1.5-fold and 4-fold; P = 0.0138 and 0.000129) and in vivo (1.7-fold; P = 0.0045). Irbesartan reduced dihydroethidium intensity (P < 0.0072) and preserved mitochondrial labeling at day 2 (P = 0.02). RNA sequencing revealed differential expression of electron transport chain genes (eg, Cox4i2 and Ndufa12), with gene set enrichment analysis showing enrichment of proinflammatory pathways in vehicle-treated explants and neuronal/metabolic pathways with irbesartan. In conclusion, irbesartan reduces oxidative stress and modulates mitochondrial and inflammatory pathways, potentially supporting RGC survival, thus providing insights relevant to glaucoma and optic neuropathies involving RGC degeneration.
This study evaluated the neuroprotective effects of inhibiting the pro-apoptotic molecule Bim (Bcl-2-interacting mediator of cell death) in retinal ganglion cells (RGCs). A mouse optic nerve crush (ONC) model was used to induce RGC degeneration. Bim expression and RGC survival were assessed by immunohistochemistry, RNA-binding protein with multiple splicing (RBPMS) immunofluorescence, and histologic analyses. Following intravitreal injection of adeno-associated virus serotype 2-shBim, retinal structure and visual function were evaluated by optical coherence tomography, flash visual-evoked potentials, and flash electroretinography. Optic nerve axons were assessed by cholera toxin subunit B-555 anterograde tracing. Primary dorsal root ganglion neurons were used to examine the effects of Bim manipulation on axon growth. RNA sequencing, real-time quantitative PCR, and pharmacologic modulation of Drd1 signaling were performed to investigate associated molecular changes. ONC induced marked up-regulation of Bim expression accompanied by substantial RGC loss. Adeno-associated virus serotype 2-shBim treatment reduced Bim expression, preserved retinal structure, increased RGC survival, partially improved visual function, and enhanced axonal preservation after injury. In dorsal root ganglion cultures, Bim knockdown promoted axon elongation, whereas Bim overexpression suppressed axon growth. Transcriptomic analysis identified significant up-regulation of Drd1 following Bim inhibition, which was confirmed by real-time quantitative PCR. Pharmacologic activation of DRD1 enhanced axon growth, whereas DRD1 blockade attenuated this effect. Taken together, these findings indicate that Bim inhibition is associated with enhanced RGC survival and axonal preservation after ONC, with Drd1-related signaling potentially contributing to these effects.
D-dopachrome tautomerase [D-DT; or macrophage migration inhibitory factor (MIF)-2] is a homolog of MIF, which shares receptor usage and downstream signaling. Although MIF is known to mediate UV-induced inflammation and matrix remodeling, the role of D-DT in cutaneous photoaging remains unclear. Therefore, this study investigated whether D-DT contributes to UVB-induced skin photoaging by modulating the expression of matrix metalloproteinase (MMP)-13. Using D-DT knockout (Ddt-/-) and wild-type (Ddt+/+) mice exposed to chronic UVB, this study assessed collagen degradation and immune cell infiltration in the skin. Also, primary dermal fibroblasts from Ddt+/+ mice were used to examine the recombinant D-DT (rD-DT)-induced MMP-13 expression and the signaling pathways. UVB exposure markedly increased the D-DT expression in epidermal keratinocytes both in vivo and in vitro. In addition to attenuated skin inflammation, Ddt-/- mice exhibited reduced UVB-induced MMP-13 expression levels, preserved type I collagen levels, and diminished leukocyte and neutrophil infiltration, comparing with wild-type mice. rD-DT directly up-regulated MMP-13 in fibroblasts, and this effect was suppressed by extracellular signal-regulated kinase and c-Jun N-terminal kinase inhibitors. Furthermore, rD-DT induced the phosphorylation of extracellular signal-regulated kinase and c-Jun N-terminal kinase in fibroblasts. In conclusion, UVB-induced skin photoaging is accelerated by the expression of MMP-13 in dermal fibroblasts following the epidermal expression of D-DT. These findings identified D-DT as a novel regulator of UVB-induced dermal remodeling and suggest its potential as a therapeutic target for photoaging.
Retinal detachment (RD) is caused by the separation of the neural retina from the retinal pigment epithelium, which leads to photoreceptor cell death as the main cause of vision loss. Here, this study investigated the role of Z-DNA-binding protein 1 (ZBP1)-mediated PANoptosis in photoreceptor death after RD and elucidated its interaction with the cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) pathway. Initially, up-regulation of ZBP1 expression in the vitreous fluid of patients with retinal detachment was detected, and retinal detachment mouse models were subsequently established via surgery. Western blot analysis and immunofluorescence confirmed a significant up-regulation of ZBP1 expression on day 3 after RD, accompanied by PANoptosis and cGAS-STING pathway activation. To investigate its mechanism, ZBP1 expression was knocked down via subretinal injection of adeno-associated virus serotype 9-shRNA-Zbp1, which was found to reduce the expression of core PANoptosis molecules and inhibit the activity of the cGAS-STING signaling. Subsequently, hematoxylin and eosin staining, transmission electron microscopy, and electroretinography confirmed that ZBP1 knockdown improved retinal morphology, partially restored outer nuclear layer thickness and retinal function, and reduced photoreceptor cell death. Similarly, i.p. injection of the cGAS-STING inhibitor C-176 attenuated ZBP1 activation and alleviated PANoptosis. In summary, ZBP1 regulates photoreceptor PANoptosis following RD, with knockdown alleviating this process via reduced cGAS-STING signaling and PANoptosis-related pathway activation.
Cardiovascular disease (CVD) is a major complication of JAK2V617F-positive myeloproliferative neoplasms (MPNs), yet the mechanisms linking mutant hematopoiesis to CVD remain incompletely understood. Three complementary murine models were developed to delineate the role of hematopoietic-endothelial crosstalk in JAK2V617F-associated CVD. In Model 1, which expresses JAK2V617F in both hematopoietic cells and endothelial cells (ECs), mice developed spontaneous dilated cardiomyopathy, right ventricular and microvascular thrombosis, arteriolar stenosis, and an increased risk of sudden death in the absence of external stressors. In Model 2, with endothelial-restricted expression, mice developed cardiac dysfunction only under a high-fat diet, accompanied by microvascular thrombosis and arteriolar stenosis, without any increased risk of sudden death. In Model 3, with mutant hematopoiesis alone, mice exhibited subtle baseline microvascular remodeling and endothelial inflammation but progressed under a high-fat diet to a phenotype resembling heart failure with preserved ejection fraction, marked by arteriolar stenosis, perivascular fibrosis, endocardial injury, and relatively preserved systolic function. Across all three models, inflammatory signaling and endothelial-to-mesenchymal transition pathways were consistently activated in cardiac ECs. Notably, endocardial ECs in Model 3 displayed pronounced transcriptional reprogramming and structural disruption. Mechanistically, thrombopoietin/MPL signaling emerged as a key mediator of hematopoietic-endothelial crosstalk; its inhibition attenuates JAK2V617F-driven cardiovascular pathology. Collectively, these findings establish a unified framework for JAK2V617F-associated CVD and identify endothelial MPL signaling as a promising therapeutic target.
Hepatocellular carcinoma (HCC) is one of the most lethal malignancies for humans. Assessing the clinical outcomes of HCC remains challenging. In this study, a panel of 20 fusion genes in 200 HCC samples was analyzed to predict the recurrence and survival rates of patients with HCC undergoing surgical interventions using machine learning models. The results showed that fusion genes, Milan criteria, serum α-fetoprotein (AFP), and pathology grade had moderate predictive accuracy for HCC recurrence. However, the combination of selected fusion genes with these clinical parameters significantly enhanced the prediction accuracy of each parameter. When models of fusion genes were applied to predict the 3-year survival rate of patients with HCC, they yielded a prediction accuracy of 72.4% in both the training and the testing cohorts. These results outperformed those from the Milan criteria (61.2% training and 58.8% testing), pathology grade (50% training and 49% testing), and serum AFP (66.3% training and 70.2% testing). The combination of a fusion gene panel with Milan criteria, pathology grade, or serum AFP level yielded significantly improved results compared to those produced by these clinical parameters alone. As a result, examining the fusion gene status of HCC samples may hold promise as a new and improved approach to assessing the clinical outcomes of this disease.
Osteosarcoma (OS), the most common primary malignant bone tumor, is characterized by a high metastatic propensity and poor survival, highlighting the urgent need to define the molecular drivers of its progression. Here, bioinformatics and clinical tissue analysis identified aberrantly high expression of IQ motif-containing GTPase-activating protein 3 (IQGAP3) in OS, with its protein level showing a significant negative correlation with patient prognosis. Functional assays demonstrate that IQGAP3 promotes OS cell migration and invasion in vitro and drives lung metastasis in vivo. Transcriptomic analysis links IQGAP3 to the focal adhesion kinase (FAK) signaling pathway. Mechanistically, IQGAP3 directly interacts with FAK and enhances its phosphorylation at Tyr397. Pharmacologic activation of FAK counteracts the suppression of migration and metastasis resulting from IQGAP3 knockdown both in vivo and in vitro. Conversely, FAK inhibition reduces phosphorylation of glycogen synthase kinase-3β (GSK-3β) at Ser9, thereby suppressing the GSK-3β/β-catenin axis. Furthermore, Wnt pathway inhibition attenuates IQGAP3-driven cell migration. Finally, the transcription factor zinc finger E-box binding homeobox 1 (ZEB1) was identified as a direct upstream regulator that binds to the IQGAP3 promoter and promotes its expression. Together, the results delineate a novel ZEB1/IQGAP3/FAK/β-catenin signaling axis that critically promotes OS lung metastasis, revealing a potential therapeutic target for intervention.
Alcohol-related liver disease (ALD) is characterized by hepatocellular injury, oxidative stress, inflammation, and extracellular matrix remodeling, yet effective therapies remain limited. Matrix-bound nanovesicles (MBVs) are bioactive vesicles embedded within the extracellular matrix that contribute to tissue repair and immunomodulation, but their role in ALD is unknown. In the current study, hepatic MBVs were isolated from mice subjected to the chronic-plus-binge ethanol model and analyzed using liquid chromatography/tandem mass spectrometry proteomics and pathway enrichment. Ethanol exposure induced marked remodeling of hepatic MBV composition, enriching proteins associated with metabolic stress, detoxification, oxidative phosphorylation, and inflammatory signaling. To investigate therapeutic potential, MBVs derived from healthy porcine urinary bladder matrix were administered to ethanol-exposed mice. MBV treatment reduced plasma alanine aminotransferase and aspartate aminotransferase levels, hepatic triglyceride accumulation, lipid peroxidation, and hepatic expression of oxidative stress-associated genes. Transcriptomic analyses showed that MBV administration partially reversed ethanol-induced pathways associated with oxidative stress, inflammatory signaling, cytokine responses, and metabolic dysfunction. MBV treatment improved hepatocellular injury despite persistent neutrophil-associated inflammatory responses, suggesting selective immunomodulation rather than broad immunosuppression. These findings identify MBVs as disease-responsive components of the hepatic microenvironment during ALD and support a proof-of-concept role for healthy extracellular matrix-derived MBVs as modulators of oxidative stress, metabolic dysfunction, and inflammatory remodeling during alcohol-induced liver injury.
Vision-language models (VLMs) represent an emerging class of multimodal artificial intelligence systems that integrate visual information with natural-language understanding and generation. In computational pathology, VLMs provide a framework for aligning histologic morphology from whole-slide images with pathology reports and other text-based knowledge sources. This review summarizes the technical foundations, major applications, evaluation strategies, and deployment considerations of pathology VLMs. Current pathology VLMs support a growing range of use cases, including image-text retrieval, label-efficient classification, visual question answering, abnormality localization, anomaly detection, report generation, and agentic workflow support. These capabilities are enabled by image encoders, text encoders or large language models, multimodal alignment strategies, and, in some systems, generative language components. Despite rapid progress, several barriers remain. The evaluation of pathology VLMs is constrained by limited domain-specific benchmarks, insufficient assessment of visual grounding, overreliance on text-based metrics, vulnerability to hallucination, and uncertain robustness under data shift. Clinical translation also requires validation across institutions, scanners, staining protocols, tissue types, and patient populations, together with workflow integration, regulatory oversight, data privacy, cybersecurity, and pathologist accountability. VLMs are therefore best viewed as assistive systems that may augment rather than replace pathologists. Responsible development will require close collaboration among pathologists, computational scientists, health systems, and regulatory stakeholders to ensure that VLMs improve pathology practice in a safe, interpretable, and clinically meaningful manner.
Effective anti-tumor immunity depends on cytotoxic T cells entering tumors, surviving there, and maintaining effector function. However, many breast cancers, particularly hormone receptor-positive subtypes, contain relatively few infiltrating T cells. This pattern reflects a hierarchy of processes, beginning with the generation of cancer-specific T cells. Once primed in lymph nodes, T cells exit nodes and enter tumors through blood vessels. This process is controlled by endothelial adhesion molecule expression and vessel integrity. Furthermore, chemokine gradients control whether T cells infiltrate cancer cell nests or are retained in the tumor stroma. Finally, the extracellular matrix adds physical and biochemical constraints through fiber alignment, cross-linking, and matricellular proteins that can limit T-cell extravasation and intratumoral positioning. These barriers are interdependent, as extracellular matrix stiffness compresses blood vessels, disrupting adhesion molecule expression and chemokine presentation. Emerging preclinical and early clinical strategies demonstrate that vascular normalization, chemokine reprogramming, and selective extracellular matrix remodeling can improve T-cell access and sensitize tumors to checkpoint blockade. Durable clinical benefit will likely require biomarker-guided combination approaches that both enable T-cell entry and preserve their function within the suppressive tumor microenvironment.
