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 ZBP1-mediated PANoptosis in photoreceptor death after RD and elucidated its interaction with the cGAS-STING pathway. Initially, upregulation 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 and immunofluorescence confirmed a significant upregulation 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 (AAV9)-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, intraperitoneal 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.

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, RBPMS immunofluorescence, and histological analyses. Following intravitreal injection of AAV2-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 CTB-555 anterograde tracing. Primary dorsal root ganglion (DRG) neurons were used to examine the effects of Bim manipulation on axon growth. RNA sequencing, qPCR, and pharmacological modulation of Drd1 signaling were performed to investigate associated molecular changes. ONC induced marked upregulation of Bim expression accompanied by substantial RGC loss. AAV2-shBim treatment reduced Bim expression, preserved retinal structure, increased RGC survival, partially improved visual function, and enhanced axonal preservation after injury. In DRG cultures, Bim knockdown promoted axon elongation, whereas Bim overexpression suppressed axon growth. Transcriptomic analysis identified significant upregulation of Drd1 following Bim inhibition, which was confirmed by qPCR. Pharmacological 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.

Histopathologic evaluation remains central to cancer diagnosis and treatment planning, yet the molecular programs underlying distinct tissue morphologies aren't routinely accessible in clinical workflows. Spatial transcriptomic/proteomic platforms provide region-specific molecular measurements but are limited by cost, throughput, and scalability. Most computational pathology models rely on either bulk tissue-based gene expression or a focused gene/protein expression-panel prediction, thereby obscuring subregion-specific morpho-molecular relationships and limiting spatial interpretation of a wider gene/protein expression network. This limitation is particularly significant in triple-negative breast cancer (TNBC), which exhibits pronounced spatial heterogeneity across tumor, stroma, and immune compartments. We developed X-SPATIO, a spatially compatible computational pipeline designed to directly link hematoxylin and eosin (H&E) morphology with region-matched mRNA and protein expression. The model was trained on H&E-defined regions of interest paired with spatially-resolved omics data obtained from GeoMx Digital Spatial Profiling. Using a multiple-instance learning approach, X-SPATIO captures morpho-molecular associations, generating spatio-morphologic attention maps that indicate predictive tissue regions. X-SPATIO demonstrated strong performance across biologically relevant spatial biomarkers, achieving area under the curve values ranging from 0.79-0.97. Attention maps revealed spatial patterns consistent with known biology, indicating alignment between learned features and tissue organization. By integrating spatial molecular ground truth with routine histopathology, X-SPATIO enables cost-effective inference of spatial biomarker expression and establishes a foundation for biologically grounded discovery and precision oncology in TNBC.

D-dopachrome tautomerase (D-DT, or MIF-2) is a homolog of the macrophage migration inhibitory factor (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, we investigated whether D-DT contributes to UVB-induced skin photoaging by modulating the expression of MMP-13. Using D-DT knockout (Ddt[-/-]) and wild-type (Ddt[+/+]) mice exposed to chronic UVB, we assessed collagen degradation and immune cell infiltration in the skin. We also used primary dermal fibroblasts from Ddt[+/+] mice 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 WT mice. rD-DT directly upregulated MMP-13 in fibroblasts, and this effect was suppressed by ERK and JNK inhibitors. Furthermore, rD-DT induced the phosphorylation of ERK and JNK 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.

Cardiovascular disease (CVD) is a major complication of JAK2V617F-positive myeloproliferative neoplasms (MPNs), yet the mechanisms linking mutant hematopoiesis to CVD remain incompletely understood. We developed three complementary murine models 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 develop spontaneous dilated cardiomyopathy, right ventricular and microvascular thrombosis, arteriolar stenosis, and an increased risk of sudden death in the absence of external stressors. Model 2, with endothelial-restricted expression, develops cardiac dysfunction only under a high-fat diet, accompanied by microvascular thrombosis and arteriolar stenosis, without any increased risk of sudden death. Model 3, with mutant hematopoiesis alone, exhibits subtle baseline microvascular remodeling and endothelial inflammation, but progresses 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 are consistently activated in cardiac ECs. Notably, endocardial ECs in Model 3 display pronounced transcriptional reprogramming and structural disruption. Mechanistically, thrombopoietin/MPL signaling emerges as a key mediator of hematopoietic-endothelial crosstalk, and 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.

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 up-regulation of Il11 transcripts in gastric mucosa. This study assessed mechanisms by which αGlcNAc deficiency induces tumorigenesis, focusing on the IL-11 receptor α (IL-11Rα)-glycoprotein 130 (gp130)-Janus kinase 2 (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 the pyloric gland cell layer was significantly decreased in A4gnt and Il11 doubly deficient (A4gnt[-/-];Il11[-/-]) mice at 10 but not 30 weeks compared with A4gnt-deficient mice, suggesting an IL-11 requirement in the early stage of tumorigenesis. Significant reduction of the proportion of phosphorylated STAT3-positive nuclei in pyloric mucosa of A4gnt[-/-];Il11[-/-] mice at 10 weeks supports the IL-11-mediated phosphorylated 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.

Alcohol-related liver disease (ALD) is characterized by hepatocellular injury, oxidative stress, inflammation, and extracellular matrix remodeling; effective therapies remain limited, however. Matrix-bound nanovesicles (MBVs), bioactive vesicles embedded within the extracellular matrix, have been implicated in tissue repair and immunomodulation in multiple injury models, but their role in ALD remains unknown. In the current study, hepatic MBVs were isolated from mice subjected to the chronic-plus-binge ethanol model and were analyzed by using liquid chromatography/tandem mass spectrometry proteomics and pathway enrichment approaches to determine how ethanol exposure alters MBV cargo. Ethanol exposure induced marked remodeling of hepatic MBV composition, with enrichment of proteins associated with metabolic stress, detoxification, oxidative phosphorylation, and inflammatory signaling pathways. To investigate therapeutic potential, healthy porcine urinary bladder matrix-derived MBVs 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 further showed that MBV administration partially reversed ethanol-induced pathways associated with oxidative stress, inflammatory signaling, cytokine responses, and metabolic dysfunction. Interestingly, MBV treatment improved hepatocellular injury despite persistent neutrophil-associated inflammatory responses, suggesting selective immunomodulation rather than broad immunosuppression. Collectively, 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.

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 osteosarcoma, 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 at Ser9, thereby suppressing the glycogen synthase kinase-3β/β-catenin axis. Furthermore, Wnt pathway inhibition attenuates IQGAP3-driven cell migration. Finally, the transcription factor 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.

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.

Neovascularization of the posterior eye is a progressive disease state, marked by an inflammatory initiation, angiogenic growth, and subsequent fibrotic degeneration of endothelial cells (ECs). Although implicated in age-related macular degeneration and proliferative diabetic retinopathy as a leading cause of irreversible vision loss worldwide, the mechanisms underpinning retinal EC dysregulation in neovascularization and fibrosis are not well understood. This study presents a transcriptomic investigation of cultured primary human microvascular retinal EC dysregulation following exposure to 10 ng/mL of six retinal neovascularization-associated signaling molecules [IL-6, tumor necrosis factor-α, transforming growth factor (TGF)-β1, TGF-β2, thrombin, and vascular endothelial growth factor-A] both individually and as a combined treatment for 24 hours. Tumor necrosis factor-α, thrombin, and TGF-β2 alone induced significant enhancement of inflammatory and angiofibrotic pathways, including phosphatidylinositol 3-kinase/Akt, NF-κB, and SMAD. BGN, CD34, COL1A2, CXCL8, IGFBP5, INHBA, SERPINE1, SNAI1, TGFB2, and TNFSF11 were identified as having overlapping, nodal roles in the pathologic dysfunction of retinal ECs. Cotreatment with all six ligands significantly enhanced differential gene expression, revealing 889 unique differentially expressed genes. Using a novel network-based gene correlation engine, GeneBunny, the cocktail group was found to mimic published retinal and choroidal EC transcriptomes from tissue of patients with age-related macular degeneration more accurately than individual treatment groups. These findings provide a biologically relevant characterization of the pathologic mechanisms driven by key retinal neovascularization-associated signaling molecules in retinal, enabling the identification of novel anti-fibrotic therapeutic targets.

Müller glia (MG) are the predominant nonneuronal cells of the vertebrate retina, playing an integral role in retinal homeostasis and photopic signal transmission. In lower vertebrates, MG retain robust regenerative potential, acting as a source of neuronal progenitors following injury. In contrast, mammalian MG primarily respond to damage with gliosis, a process that provides initial neuroprotection but ultimately restricts their neurogenic capacity. This limitation is especially consequential in humans, where genetic disorders and age-related diseases, such as diabetic retinopathy, glaucoma, and macular degeneration, lead to retinal neuron death and irreversible visual loss. One promising therapeutic strategy is to reprogram MG in vivo into stem-like progenitor cells capable of replacing lost neurons, with the expectation that these new cells can form functional synaptic circuits, replace lost neurons, and restore retinal function. Recent studies have reviewed the integrated regulatory network, encompassing signaling pathways, including Notch, Wnt/β-catenin, mitogen-activated protein kinase/extracellular signal-regulated kinase, phosphatidylinositol 3-kinase/AKT, SMADs, and Hippo-YAP, alongside key transcription factors and epigenetic mechanisms, which collectively govern MG cell cycle re-entry and neurogenesis in mammals. This review synthesizes current knowledge of this integrated regulatory network, mainly focusing on the role of signaling pathways in MG regeneration, highlighting strategies to harness the intrinsic neurogenic potential of MG for retinal repair, underlining their promise as cellular targets for curing degenerative retinal diseases from intrinsic cellular sources.

Organ and tissue functions emerge from the coordinated activity of cell networks. Therapeutics that act on pathogenic cell networks, modulating their cellular interplay, follow naturally. Over several decades, our laboratory has developed a series of approaches for rewiring cell networks, culminating in a class of cell surface-directed signal converter proteins (SCPs) that do so by modulating juxtacrine and autocrine signaling in and among their nodal cells. A first such SCP has now produced encouraging clinical data for cancer immunotherapy. Yet, these early network-directed fusion proteins rest on a deliberately simplified picture: discrete end-cell types plugged into graphically tractable networks. That picture is increasingly at odds with what computational cell typing and spatiotemporal analytics, along with epigenetics, now reveal-a hyperdiverse, plastic, experience-shaped cellular landscape embedded in dynamic, multiway networks. Setting the stage for a next generation of network modulators, an extended cell differentiation synthesis, is proposed, which formalizes paracell ultradifferentiation and aging-associated differentiation phases. According to this model, cells are ever evolving, and no two cells are alike. A richer cellular ontology forces a more elaborate network ontology, with paralogous networks and their shifting subnetworks opening a concrete design space for next-generation network-directed SCP therapeutics. This exploration calls for a willingness to embrace complexity more fully and borrow freely from conceptual fields close and afar.

Liver X receptor alpha (LXRα) regulates serum cholesterol and lipoprotein levels under homeostatic conditions and in response to dietary fat. Previously, aged Lxrα[-/-] mice fed a standard diet were shown to develop phenotypes resembling early age-related macular degeneration (AMD). Herein, the hypothesis tested was that introducing a high-fat, high-cholesterol (HFC) diet in young Lxrα[-/-] mice could bypass aging and accelerate onset of AMD-like pathology. Young Lxrα[+/+] and Lxrα[-/-] mice (2 to 3 months old) were fed either a standard diet or an HFC diet for up to 2 months. Retinal function and morphology were assessed by electroretinography and fundus/optical coherence tomography imaging, histopathology by transmission electron microscopy, and retinal markers by immunostaining. Young Lxrα[-/-] mice fed an HFC diet exhibited impaired retinal function with reduced scotopic a- and b-waves, photopic b-waves, and c-waves. Although the retinal ultrastructure of control and HFC-fed Lxrα[+]/[+] mice appeared normal, HFC-fed Lxrα[-/-] mice showed retinal pigment epithelium thinning, disorganized basal infoldings, intracellular lipid droplets, thin basal laminar deposits, and Bruch membrane thickening enriched in apolipoprotein E. Increased microglia/macrophage infiltration in the retina and choroid suggested elevated local inflammation. Although an HFC diet accelerated AMD-like changes in young Lxrα[-/-] mice, these lesions were less severe than those observed in aged Lxrα[-/-] mice on a standard diet, indicating that aging remains the dominant driver of disease severity.