Chronic wound management requires advanced dressings with sufficient adhesive properties and mechanical stress. Glycyrrhizic acid (GA)-derived hydrogels hold remarkable potential as biomaterials for diverse wound healing, however, their poor mechanical performance, limited stability, and inevitable cytotoxicity at high gelling concentrations severely restrict in vivo applications. Here, an innovative charred Trachycarpus-derived carbon dots (CT-CDs)-linked GA hybrid hydrogel (CT@GA-gel) was fabricated and imparted in injectable and self-healing properties for comprehensive therapy of diabetic wounds. Specially, the addition of CT-CDs with negative charge enabled the GA crosslinking to form hydrogels at very low concentrations (0.5% GA). Meanwhile, CT-CDs could significantly improve the mechanical properties and confer tissue adhesion of GA hydrogel for rapid hemostasis. Benefiting from the ROS scavenging activity of CT-CDs, the CT@GA-gel achieved immune microenvironment regulation, re-epithelialization and hair follicle hyperplasia, thereby facilitating chronic wound closure. Using transcriptomics analysis, we confirmed that the CT@GA-gel efficiently increased the gene expression associated with hemostasis, cell adhesion and extracellular matrix deposition, indicating the enhanced proliferation and remodeling during wound repair process. In the field of regenerative medicine, this work brings hope for the treatment of chronic tissue injury.

During early embryogenesis, the uterine environment undergoes marked biophysical changes that guide the embryonic cell fate. However, replicating these stage-specific cues in vitro remains challenging. Herein, we introduce a gelatin-based coacervate matrix with phase-transition-mediated tunable mechanics to recapitulate the biophysical cues of pre- and peri-implantation stages. Driven by reversible hydrophobic interactions, liquid-liquid phase separation produces coacervates with ultradynamic structures that enable dramatic volume expansion during cell proliferation in preimplantation stage. Furthermore, the liquid-like coacervate emulates the loosely organized immature extracellular matrix (ECM) of the uterine fluid environment, providing moderate cell-matrix interactions that preserve stemness. Additionally, the coacervate-solution transition allows the efficient harvesting of highly viable embryonic stem cell colonies. Moreover, upon stiffening through the coacervate-hydrogel transition, the matrix promotes peri-implantation-like invasive behaviors, including enhanced cell-matrix adhesion and secretion of ECM-degrading enzymes. These findings establish the biomimetic coacervate matrix as a versatile platform for clonal growth, stemness maintenance, and lineage initiation, offering new opportunities for developmental modeling and therapeutic applications.

Enamel mineralization critically depends on maturation-stage ameloblasts (M-ABs) regulating pH, protein secretion, and cell-matrix adhesion. However, the molecular mechanisms underlying these processes remain poorly understood. This study identifies the vacuolar-type H+-ATPase (V-ATPase) a3 subunit as a key regulator of enamel formation via its role in secretory lysosome trafficking. In a3 knockout (a3KO) mice and cultured ameloblasts, a3 is required for both lysosomal acidification and the directional transport of odontogenic ameloblast-associated protein (ODAM)-containing secretory lysosomes to the ruffled border membrane of M-ABs. At this site, ODAM is crucial for mediating ameloblast adhesion to the enamel matrix. Loss of a3 caused severe enamel hypomineralization, characterized by reduced matrix acidification, cystic enamel defects, abnormal ruffled border morphology, and ameloblast detachment from the mineralizing surface. In vitro, a3-deficient ameloblasts exhibited significantly impaired adhesion to hydroxyapatite, decreased ODAM expression, and suppressed lysosomal acidification, indicating a3 is functionally required for maintaining ameloblast function and polarity. Mechanistically, Rab27A served as an important adaptor linking a3-positive secretory lysosomes to the microtubule network, enabling their polarized movement toward the distal plasma membrane. Disruption of this a3-Rab27A axis in a3KO cells mislocalized secretory lysosomes and defective ODAM delivery into the enamel matrix, compromising enamel mineralization. These findings reveal a new mechanism by which a3 orchestrates lysosomal positioning and ODAM secretion in enamel-forming cells. By integrating proton transport with vesicular trafficking and adhesion protein delivery, a3 functions as a key regulator of enamel mineralization. This study provides new insights into the pathogenesis of enamel hypomineralization and identifies a3 and its associated pathways as potential therapeutic targets for treating developmental enamel defects.

Dependence of mesenchymally transitioned tumor niche fitness on cell-cell and cell-matrix adhesions.

TAGLN is identified as a key prognostic gene in colorectal cancer through TCGA-COAD analysis, with high expression correlating with poor survival and advanced tumor stage. Functionally, TAGLN overexpression promotes epithelial-mesenchymal transition and enhances cancer cell migration. Transcriptomic profiling reveals its involvement in extracellular matrix remodeling and cell adhesion pathways. Mechanistically, TAGLN expression is upregulated by stiff tumor-mimicking extracellular matrix and is correlated with key mediators of collagen crosslinking and EMT. Clinically, TAGLN exhibits progressive overexpression from normal epithelium to primary tumors and metastatic lesions. These findings establish TAGLN as a stiffness-responsive regulator of ECM remodeling and EMT, driving colorectal cancer metastasis and serving as a potential therapeutic target.

The development of biomaterials provides a promising avenue for uncovering pathogenic mechanisms and exploring novel therapeutic strategies, particularly for refractory dermatoses, such as prurigo nodularis (PN), a chronic pruritic dermatosis of unknown etiopathogenesis. Although immune cells play important roles in skin inflammation, few studies have investigated the pathogenesis of PN from the perspective of the interaction between basal layer epidermal stem cells (EpSCs) and the basement membrane (BM) extracellular matrix (ECM). In this study, we used quantitative proteomic techniques to construct protein expression profiles of EpSCs and their extracellular vesicles from patients with PN. By combining protein‒protein interaction networks and pathological analyses, we investigated the relationships among EpSCs, EpSC-derived exosomes, and the immune microenvironment of PN. EpSCs exhibited abnormal morphology in patients with PN, with EpSC dysfunction mediated by the activation of inflammasome pathways. To further analyze these mechanisms, we employed biomaterial-based models, including multiple models of tissue-engineered skin and 3D epidermal organoids. These results revealed that regulating cell‒matrix adhesion between basal layer EpSCs and the BM ECM promoted the function of EpSCs, making this regulation a potential target for the treatment of PN EpSC dysfunction.

The choroid plexus (CP) regulates barrier integrity, cerebrospinal-fluid (CSF) dynamics, and immune surveillance, yet its role in Alzheimer's disease (AD) remains poorly defined. We performed snRNA-seq on CP samples from 69 ROSMAP participants spanning normal cognition, mild cognitive impairment, and AD dementia, and integrated these data with spatial transcriptomics, snATAC-seq, and proteomics from CP tissue and CSF. We identified 17 CP cell states and uncovered widespread disease-associated transitions that converged into three major phenotypic axes. Along the inflammatory axis, epithelial cells and border-associated macrophages (BAMs) showed progressive immune activation, with BAMs shifting from inflammatory to stress-dominant states. In the barrier axis, epithelial cells, fibroblasts, and endothelial cells exhibited reduced junction-related gene expression and broad alterations in transport pathways. Epithelial cells also showed late-stage cilia loss and CSF-regulatory pathway impairment, indicating a breakdown in epithelial polarity and CSF sensing, consistent with abnormal CSF proteomic signatures. Along the remodeling axis, fibroblasts showed ECM alterations, while epithelial and stromal cells demonstrated aberrant cell-matrix adhesion pathways. Spatial neighborhood analysis revealed a multicellular signaling hub, with epithelial-rich niches showing the strongest dysregulation in AD. Together, these findings define a unified model of CP dysfunction in AD and position the CP as an active, multicellular contributor to AD pathophysiology.

BackgroundHepatocellular carcinoma (HCC) is a common malignant tumor worldwide, with complex pathogenesis and poor clinical prognosis. It was shown that hepatocyte nuclear factor 1β (HNF1B) is strongly expressed in hepatocyte and involved in liver development. Relevant studies have shown that HNF1B can be used as a marker for the identification of hepatocellular cholangiocarcinoma. However, regarding the mechanism by which HNF1B regulates the behavior of cancer cells to influence the development, malignancy and metastasis ability of HCC, is poorly understood at present.MethodsIn this study, the proliferation, migration, invasion, and apoptosis of liver cancer cells was investigated by modulating the expression of HNF1B in HCC cells. Furthermore, bioinformatics analysis: including differential gene and survival analysis, clinical relevance, GO/KEGG enrichment, immune cell infiltration, gene pathway prediction and PPI network analysis were applied to explore the mechanism of HNF1B to influence the development, malignancy and metastasis of HCC.ResultsOur results indicated that over-expression of HNF1B promoted the proliferation (*P < 0.05), migration (***P < 0.001) and invasive (**P < 0.01) ability of HCC cells, while significantly inhibited apoptosis. Importantly, overexpression of HNF1B may be associated with the transformation process from hepatocellular carcinoma (HCC) to intrahepatic cholangiocarcinoma (ICC) (****P < 0.0001). Bioinformatics analysis showed that HNF1B increased tumor cell heterogeneity by remodeling in the tumor microenvironment including immune cell infiltration, cell adhesion, and extracellular matrix. Moreover, HNF1B regulates the transformation of HCC to ICC through Notch, Hippo and neuroactive ligand-receptor interaction signaling pathways.ConclusionsOur study elucidated the functional roles of HNF1B on regulating the biological behavior of HCC cells, suggesting that HNF1B could be an important molecular marker of HCC. This finding provides a potential molecular target for the diagnosis and prognosis of HCC.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12885-025-15408-x.

Aims Epidemiological studies indicate a heightened risk of pancreatic adenocarcinoma (PAAD) in patients with type 2 diabetes mellitus (T2DM). This study investigates the molecular mechanisms underlying their comorbidity. Materials and methods Common DEGs between T2DM and PAAD were identified from GEO datasets. Functional and pathway enrichment analyses were performed via PPI, GO, and KEGG. Core genes were screened and their diagnostic value was validated by ROC curves. Immune infiltration and TF–mRNA–miRNA regulatory networks were constructed to explore disease mechanisms. Core gene expression and prognostic significance in PAAD were assessed using GEPIA2 and HPA. Potential therapeutics targeting core genes were predicted via the Therapeutic Target Database. Results A total of 35 DEGs were identified. GO analysis linked these genes to cell adhesion and extracellular matrix (ECM) components. KEGG enrichment highlighted ECM-receptor interaction as the top pathway. Key ECM-related molecules—ITGA3, FN1, LAMB3, ITGA2, and LAMC2—were upregulated in both T2DM and PAAD. Six potential therapeutic agents targeting ITGA2, LAMB3, and FN1 were identified. Conclusion Three genes and associated known drugs identified in this study may serve as potential targets for treating the coexistence of the two diseases.

Quercetin alleviates intervertebral disc degeneration by disrupting CEBPB-HMGA1 interaction to suppress TNFA production and reduce ECM degradation.

Background and PurposeBreast cancer presents a substantial clinical challenge because of its complex aetiology and diverse phenotypic presentations. ST6Gal1 expression and epithelial-to-mesenchymal transition (EMT) frequently lead to metastasis, drug resistance and poor prognosis in many cancers. Nevertheless, the molecular details surrounding ST6GAL1 in the carcinogenesis of breast cancer, especially in the EMT of breast cancer, remain unclear. The objective of this study was to clarify the possible role and mechanism of ST6GAL1 in breast cancer.MethodsPCR, WB, and IHC were employed to analyze the expression of ST6GAL1, epithelial-mesenchymal transition (EMT) markers, and components of the HIF-HK2 signaling pathway in MCF-10A, MDA-MB-231, and MCF-7 cells. Wound-healing, cell adhesion, drug resistance and extracellular matrix invasion assays were used to analyse the effects of ST6GAL1 on the biological process of breast cancer cells. The HIF-HK2 signalling pathway was also analysed.ResultsST6GAL1 expression is increased in breast cancer. Altered expression of ST6GAL1 affects the biological function of breast cancer cells both in vitro and in vivo. ST6GAL1 knockdown inhibited EMT in breast cancer cells. ST6GAL1 Mediates the Activity of the HIF-HK2 Signalling Pathway in Breast Carcinoma Cells.ConclusionIn our study, in vitro and in vivo models revealed that ST6GAL1 promotes malignant phenotypes in breast cancer cells and regulates the EMT process through activation of the HIF-HK2 signalling pathway.

Comparative analysis of antibody-mediated loss-of-function versus gene knock-out and knock-down.

Distinct plasma proteome in severe pulmonary hypertension associated with chronic lung disease.

Abstract Background Alzheimer's disease (AD) is a complex neurodegenerative disorder characterized by cognitive decline and disruptions in neurovascular integrity, inflammation, immune signaling, and synaptic connectivity. While single‐cell and bulk RNA‐seq have revealed cell‐type‐specific changes in AD, the intercellular communication networks driving these processes remain unclear. This study introduces a novel integrative pipeline to systematically identify and prioritize robust cell‐cell interactions contributing to AD pathology. Method We developed an advanced computational pipeline (Figure 1) that, to our knowledge, is the first to integrate cell prioritization tools with cell‐cell interactions to identify AD‐associated interactions. To achieve this, we integrated RNA‐seq from the Mount Sinai Brain Bank (MSBB) and single‐cell RNA‐seq (scRNA‐seq) from Grubman, Mathys, and MIT datasets, totaling over 2.4 million cells. The pipeline combined Diagnostic Evidence GAuge of Single cells (DEGAS), a transfer learning framework for identifying AD‐associated cellular states, with CellChat, which infers intercellular communication networks by mapping ligand‐receptor interactions across cell types. To ensure robustness, correlation analysis validated ligand‐receptor co‐expression, while differential gene expression (DEG) and gene ontology (GO) analyses identified pathways linked to AD‐associated states. Consistent interactions across datasets will be experimentally validated via colocalization using immunofluorescence (IF) and binding via co‐immunoprecipitation (Co‐IP). Result Our pipeline consistently identified cell‐cell interactions underlying AD pathology across datasets (Figure 2). FN1‐SDC4 interactions between endothelial cells and astrocytes were associated with blood‐brain barrier dysfunction (Figure 3). CADM1‐CADM1 interactions, observed between oligodendrocytes and oligodendrocyte progenitor cells (Figure 3), highlighted their role in synaptic organization and connectivity deficits. The BSG‐PPIA interaction, specific to endothelial cells, was linked to vascular inflammation (Figure 3). Additional interactions, JAM3‐JAM3 and PTPRM‐PTPRM, were consistently identified in control contexts, suggesting roles in synaptic maintenance and axonal guidance. Correlation analysis further validated ligand‐receptor co‐expression, while DEG and GO analyses highlighted enriched pathways, including extracellular matrix organization, cell adhesion, and synaptic signaling. These results are now being further validated via IF and Co‐IP. Conclusion This study highlights the novelty of our pipeline in integrating RNA‐seq data to identify robust intercellular communication networks in AD. By prioritizing consistent interactions across datasets, we provide insights into AD pathology that inform experimental validation and therapeutic development.

A major challenge in cardiac research is the limited translatability of drug screening and toxicity assays due to the use of in vitro models that poorly mimic the native cardiac environment. Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) offer a promising route forward, but conventional 2D culture on rigid substrates hinders their functional maturation and predictive accuracy. This study addresses this problem by investigating the effect of hybrid fibronectin-based hydrogels with tunable stiffness on the mechanical and electrical properties of hiPSC-CMs. We engineered hydrogels with stiffness mimicking the lowest range of neonatal heart tissue stiffness (2-4 kPa) and compared hiPSC-CM behavior on these substrates to that on standard fibronectin-coated glass. Our results demonstrate that hydrogel culture promotes more uniform and stable cardiomyocyte contractions, as evidenced by increased single peak percentages and altered contraction duration. Electrophysiological analysis revealed that hydrogel stiffness influences action potential duration and signal amplitude. Furthermore, hiPSC-CMs on hydrogels exhibited enhanced cell-matrix and cell-cell adhesion, indicating improved structural and functional connectivity. Drug testing with known cardioactive compounds, including isoproterenol and nifedipine, revealed distinct differences in drug responses between hydrogel and glass cultures, suggesting that hydrogels provide a more physiologically relevant platform for assessing drug effects. This work highlights the potential of engineered hydrogel substrates to enhance the functional maturity and predictive accuracy of hiPSC-CMs for cardiac research and drug development.

Colorectal cancer (CRC) stands as the most common gastrointestinal malignancy, with the Follistatin-like 1 (FSTL1) gene associated with unfavorable outcomes in diverse cancer forms. Nonetheless, the precise mechanisms through which FSTL1 influences CRC pathogenesis and its associated multi-omics characteristics remain unclear. This study leveraged RNA sequencing and single-cell RNA sequencing (scRNAseq) data from the Gene Expression Omnibus (GEO) database to scrutinize the expression profile, prognostic significance, and clinicopathological relevance of FSTL1 in colorectal cancer. Gene set enrichment analysis (GSEA) and protein-protein interaction (PPI) network analysis were conducted to delve into the potential biological functions of FSTL1 in colorectal cancer and its associated pathways. The ESTIM algorithm and Tumor Immune Estimation Resource (TIDE) database were utilized to investigate FSTL1 expression in immune cell infiltration and immune checkpoints. Mutation characteristics of FSTL1 were elucidated using the cBioPortal database. The pRRophetic package was employed to identify potential chemotherapeutic drugs targeting FSTL1. Expression levels of FSTL1 in colon cancer and adjacent normal tissues were evaluated using the Human Protein Atlas (HPA) database. Furthermore, single-cell sequencing technology and cell communication were employed to assess the immunogenomic features of FSTL1 in the tumor microenvironment. In vitro experiments were conducted to validate the efficacy of FSTL1. FSTL1 levels were markedly higher in colorectal cancer tissues compared to healthy tissues, correlating significantly with unfavorable patient outcomes. Elevated FSTL1 expression was closely linked to key biological processes including the PI3K pathway, cell adhesion, proliferation, migration, and extracellular matrix remodeling. Furthermore, FSTL1 expression exhibited strong associations with immune cell levels and the tumor immune microenvironment in colorectal cancer. Notably, FSTL1 mutations predominantly comprised missense mutations and displayed significant correlations with various immune checkpoints and low methylation levels. Axitinib emerged as a promising targeted therapeutic option for patients with high FSTL1 expression. Analysis from the HPA database confirmed elevated FSTL1 expression in colorectal tissues compared to adjacent normal tissues. Single-cell sequencing analysis identified stromal cells as the primary source of abundant FSTL1 expression, potentially influencing tumor microenvironment remodeling by myeloid and B cells via the APP-CD74 pathway. Downregulating FSTL1 expression in colon cancer cells suppresses the proliferation, motility, and invasiveness of SW480 and HCT116 colon cancer cells. In summary, the findings of the study indicate that elevated FSTL1 expression could serve as a prognostic biomarker for unfavorable outcomes in colorectal cancer (CRC) diagnosis and may also identify potential targets for immunotherapy in CRC.

Background:In contrast to what has already been shown in HFpEF associated left ventricular (LV) diastolic dysfunction, leukocytes’ role in frequently occurring right ventricular dysfunction (RVD) secondary to HFpEF are so far missing, partially due to the lack of suitable small animal models. Here, we follow a translational research approach by establishing a murine HFpEF model developing manifest RVD and analyzed human HFpEF cohorts to study the mechanistic link between leukocytes and RVD in HFpEF.Methods:8-week-old male and female C57BL/6J or Cx3cr1CreER/+R26tdTomato/+ mice were divided into four experimental groups: i) chow, ii) HFpEF (N[ω]-nitro-l-arginine methyl ester (L-NAME), 60% high-fat diet), iii) chronic hypoxia (10% O2) and iv) HFpEF and hypoxia (RV-HFpEF) to assess bi-ventricular function and myeloid cell dynamics. To test whether myeloid cells are causally involved in the development of RV remodeling in HFpEF, we additionally treated RV-HFpEF mice with the colony stimulating factor 1 receptor inhibitor PLX-5622 (PLX) to deplete myeloid cells. After 12 weeks, all experimental groups were subjected to transthoracic echocardiography, invasive hemodynamics or flow cytometry.Results:RV-HFpEF resulted in LV diastolic dysfunction indicated by increased E/E’ ratio, reduced global longitudinal peak strain, smaller end-diastolic diameters and increased isovolumetric relaxation time compared to chow. RV-HFpEF animals developed RV hypertrophy and RVD evident as increased Fulton’s index and collagen content as well as elevated RV systolic pressures (RVSPs) and reduced tricuspid annular plane systolic excursion, respectively. Flow cytometric analyses revealed elevated total leukocyte, monocyte, and macrophage counts in RV tissue of RV-HFpEF compared to chow or LV tissue from RV-HFpEF animals. These data were confirmed by unbiased proteomic analyses of RV tissue from RV-HFpEF mice, demonstrating increased abundance of proteins involved in activation of the innate immune system, macrophage chemotaxis, cell adhesion and extracellular matrix organization when compared to LV tissue or other experimental groups. Fate mapping experiments revealed that recruited monocyte-derived macrophages became the main source of total cardiac macrophages in RV tissue from RV-HFpEF mice. Depletion of myeloid cells was associated with rescued RVSP profiles compared to RV-HFpEF control mice. In HFpEF patients, RV dilation was associated with an increased percentage of circulating monocytes. In RV biopsies from HFpEF patients, we found increased expression of adhesion molecules, fibrotic markers and inflammatory transcripts.Conclusion:We demonstrate that dysregulated myeloid cell dynamics are associated with, and directly contribute to, the pathogenesis of HFpEF-associated RVD in humans and mice.

Primary cilia have emerged as key regulators in cancer biology, influencing tumor progression and therapeutic response through diverse signaling pathways. In this study, we identify WDR60, a component of the dynein-2 complex essential for retrograde intraflagellar transport, as a novel modulator of pancreatic neuroendocrine tumor (Pa-NET) behavior. Transcriptomic analysis of the GEO dataset GSE73338 revealed that WDR60 is significantly upregulated in both primary and metastatic Pa-NETs compared to normal pancreatic islets. Moreover, WDR60 expression is higher in G2 compared to G1 Pa-NETs. Functional analyses in QGP-1 cells following WDR60 silencing demonstrated broad transcriptional reprogramming with enrichment of pathways related to cell adhesion and extracellular matrix (ECM) remodeling. Notably, WDR60 knockdown reduced cell migration and enhanced adhesion without affecting cell viability or proliferation. Among the key upregulated genes were PIK3AP1, RAP1B, and RFLNA, suggesting that WDR60 is involved in regulating PI3K/AKT signaling and cytoskeletal dynamics. To explore potential therapeutic implications, we examined the effects of Ciliobrevin A (HPI-04), an inhibitor of Hedgehog signaling and ciliogenesis. HPI-04 significantly reduced WDR60 expression, impaired cell migration, and increased adhesion. RT-qPCR confirmed overlapping gene expression changes between HPI-04 treatment and WDR60 silencing, although some differences, such as CD164 regulation, suggest WDR60-independent mechanisms. Collectively, these findings identify WDR60 as a critical regulator of cell motility and adhesion in Pa-NETs via ciliary signaling and cytoskeletal remodeling. They also support the therapeutic potential of targeting cilia-associated pathways, including WDR60 and Hedgehog signaling, in the treatment of Pa-NETs.

Background: Adolescents and young adults (AYA) with papillary thyroid carcinoma (PTC) often present with more extensive cervical lymph node metastasis (LNM) than older adults (AD). We aimed to identify age-associated molecular and immune features that might explain this phenotype and to explore potential translational implications for managing aggressive AYA PTC. Methods: We analyzed clinical and transcriptomic data from 501 PTC cases in The Cancer Genome Atlas (TCGA), stratified as AYA (<30 years, n = 64) and AD (≥30 years, n = 437). An institutional RNA-seq cohort (n = 13; 7 AYA, 6 AD) was used to screen for differentially expressed genes (DEGs). DEGs were defined by p ≤ 0.05 and |log2 fold change| ≥ 1. Intersection with invasion- and dissemination-related gene sets yielded a final age-related DEG list. Functional enrichment (GO/KEGG via DAVID), PPI network analysis (STRING, Cytoscape/cytoHubba), and immune deconvolution (CIBERSORT LM22) were performed. Protein-level validation was carried out by immunohistochemistry (IHC) in an independent cohort (n = 56; 28 AYA, 28 AD). Statistical comparisons used chi-square/Fisher's exact tests for categorical variables, t-tests or nonparametric tests for continuous variables, and EdgeR with FDR correction for transcriptomic analyses. Results: In TCGA, LNM was more frequent in AYA than in AD (62.1% vs. 47.8%, p = 0.031). From intersected analyses, we identified 239 core DEGs distinguishing highly invasive, age-related tumors. Key upregulated genes in AYA included CXCR4, OPCML and S100A2; downregulated genes included ATP1A3, CHL1, HLA-DRA and IL-1β. Enriched pathways involved extracellular matrix organization, cell adhesion, calcium signaling and canonical oncogenic cascades (PI3K-Akt, MAPK, Wnt, Ras). Immune deconvolution showed reduced naïve B cells, M1 and M2 macrophages and resting mast cells and an increased proportion of M0 macrophages in AYA tumors. IHC validated differential protein expression for seven markers. Collectively, the data indicate an immune-suppressed, immune-excluded microenvironment in AYA PTC. Conclusions: AYA PTC exhibits distinct molecular and immune features that may underlie its propensity for lymphatic dissemination. These findings support evaluation of translational strategies, such as CXCR4 inhibition, restoration of antigen presentation, and macrophage reprogramming, to convert "cold" tumors into immune-permissive lesions. Validation in larger, prospective, multicenter cohorts is required.

Integrin-mediated cell-matrix adhesions regulate communication between cells and the extracellular matrix. In matrix-secreting cells, fibrillar adhesions (FBs) containing high levels of α5β1 integrins and the tensin3 adaptor protein are essential for fibronectin (FN) fibrillogenesis. Here, we demonstrate that tensin3 binds to four helical regions (R3, R4, R8, and R11) of talin, the principal integrin activator. Structural analysis revealed the residues critical for the tensin3-talin interaction, and mutational analysis showed that talin R8 and R11 are essential for FB formation and FN fibrillogenesis. Cellular experiments demonstrate that tensin3 binding to talin not only regulates integrin activation, but also modulates tensin3's propensity to undergo liquid-liquid phase separation (LLPS). Formation of such LLPS condensates increased when cells were plated on soft substrates compared with stiff ones. This effect was abolished by blocking the interaction between tensin3 and talin. Our data suggest a model in which LLPS condensates provide a signaling platform involved in cellular responses to sudden changes in tissue mechanics.