Search Results (148)

Canine mammary cancers (CMCs) are one of the most prevalent types of neoplasm in dogs, are frequently malignant, and display high tumour heterogeneity, making evaluating prognosis and predicting successful treatment outcomes difficult. In a previous pilot study, overexpression of the Wnt pathway-associated protein SFRP1 was found to correlate with negative metastasis status in CMCs at both mRNA and protein levels. To establish SFRP1 as a potential biomarker for CMC progression, additional verification of these results in an independent dataset is required, as well as an investigation as to whether SFRP1 expression in CMCs is associated with altered Wnt- or RANKL signalling pathways. In an independent verification cohort of 122 cases of archival CMC FFPE material, expression of SFRP1 was assessed by RT-qPCR and immunohistochemistry. The same tumours were further assessed for RANKL, phosphoROCK2, and NFkB-p65 protein expression. Our data verified that SFRP1 mRNA (p = 0.025) was negatively associated with metastasis status; however, differences in protein expression did not reach statistical significance (p = 0.139). Neither did SFRP1 significantly correlate with expression of any of the other proteins tested. Instead, a strong association was found for RANKL positivity with increased metastasis status (p < 0.001). Co-expression of SFRP1 significantly lowered the higher risk of metastatic spread when compared to RANKL_pos/SFRP1_neg CMCs (p = 0.033). Noticeably, all vascular-invasive cell clusters observed in tissue section vessels stained positive for RANKL. Our study identified RANKL expression as a strong marker for cancer progression with a strong link to vascular-invasive cells. However, SFRP1 expression may potentially suppress the pro-metastatic nature of RANKL_pos CMCs. Full article

Obesity disrupts skeletal muscle metabolism through insulin resistance, oxidative stress, and ectopic fat deposition, yet transcriptomic findings across individual studies remain inconsistent. We performed a meta-analysis of four independent RNA sequencing (RNA-seq) studies of human vastus lateralis muscle, comparing 29 individuals with obesity (body mass index (BMI) ≥ 30 kg/m²) and 23 with normal weight. Differential expression was analyzed using DESeq2, with age and sex included as covariates in studies where individual-level data were available. Study-level results were integrated using the direction-aware inverse normal method (weighted Stouffer). Between-study heterogeneity was assessed by gene-level I² statistics derived from random-effects meta-analysis of log₂ fold changes. Functional annotation was performed with Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. The weighted Stouffer method identified 2136 differentially expressed genes (DEGs) (adjusted p < 0.05), comprising 1028 upregulated and 1108 downregulated genes, of which 674 (31.6%) were detected only through the meta-analysis. Three genes—PHLDA3 (down), CNKSR2 (down), and SFRP4 (up)—were significant in every individual study and in the combined analysis. Downregulated DEGs were enriched in cytoplasmic translation, ribosomal structure, and oxidative phosphorylation, whereas upregulated DEGs were associated with extracellular matrix organization and the focal adhesion pathway. This RNA-seq meta-analysis of skeletal muscle in obesity identifies robust DEGs and dysregulated pathways, providing candidate targets for future mechanistic and translational research. Full article

Intervertebral disk pathology, including disk herniation and degeneration, is a major contributor to chronic low back pain, and when conservative treatment fails, surgical management often involves discectomy-based procedures that leave residual annulus fibrosus (AF) defects associated with reherniation and progressive degeneration. These limitations have motivated interest in regenerative strategies using biomaterial scaffolds; however, reproducing the hierarchical, angle-ply architecture of the AF remains challenging. Here, we present a single-step extrusion-based 3D-printing approach to fabricate polycaprolactone (PCL) scaffolds with aligned microscale surface grooves that promote AF-like organization. Patterned nozzles with circumferential peaks generated uniaxial concave microgrooves (10–17 µm wide) directly during printing, enabling formation of multilamellar angle-ply constructs. Human bone marrow-derived mesenchymal stem cells cultured on patterned scaffolds aligned longitudinally within concave grooves, forming end-to-end arrays that guided extracellular matrix deposition. Gene expression analysis showed that topographical cues governed cellular organization without significantly altering gene expression profiles, while TGF-β3 supplementation upregulated outer AF-associated markers, including COL1, COL12, SFRP2, MKX, MCAM, and SCX. TAGLN expression increased specifically on patterned scaffolds in the absence of TGF-β3, indicating an association between microgroove-guided cellular organization and TAGLN expression, warranting further investigation into potential tension-related mechanisms. This novel single-step extrusion-printing approach leverages custom nozzle geometry to impart concave microgrooves, facilitating scalable fabrication of multilamellar angle-ply scaffolds that induce aligned cellular organization and support potential applications in annulus fibrosus repair, as well as mechanobiological studies of anisotropic musculoskeletal tissues. Full article

Extrachromosomal circular DNA (eccDNA) has emerged as a pivotal contributor to cancer progression, facilitating oncogene amplification, dysregulated gene expression, and tumor heterogeneity. Despite its significance in cancer, the interplay between eccDNA and key epigenetic regulators such as EZH2 remains largely unexplored. In this study, we systematically investigate the correlation between Tazemetostat, a highly selective EZH2 inhibitor, and alterations in the eccDNA landscape and transcriptional programs in ovarian cancer. Through integrated profiling using Circle-seq and RNA sequencing, we demonstrate that EZH2 inhibition is associated with markedly reprogrammed eccDNA dynamics. Furthermore, multi-omics integration identified that 67 genes exhibited concordant changes in both eccDNA abundance and transcript expression. Subsequent analyses also pinpointed 11 genes as putative effectors of drug response. Notably, spatial single-cell transcriptomics identified SFRP1 as the most consistently reactivated tumor suppressor across eccDNA, bulk expression, and spatial datasets, based on predefined statistical and biological criteria, by Tazemetostat. Moreover, SFRP1 was one of the genes that varied the most within cancer-associated fibroblast populations, exhibiting distinct spatial expression patterns. Taken together, this study establishes the first potential evidence that EZH2 inhibition may reprogram eccDNA dynamics to potentially restore SFRP1 tumor suppressor expression in ovarian cancer. By integrating multi-omics and spatial single-cell transcriptomics, we uncovered a novel epigenetic–eccDNA axis that may contribute to oncogenic plasticity and therapeutic resistance. This could result in a paradigm shift in targeting eccDNA-driven malignancies. Full article

Androgen receptor (AR) signaling plays a key role in male pattern baldness. We investigated whether targeting Dickkopf 1 (DKK1) and Secreted frizzled-related protein 1 (SFRP1), two AR-regulated genes, offers a novel therapeutic strategy for hair loss. AR expression was validated in freshly frozen human scalp hair follicles (HFs). AR knockdown was induced in human HFs using AR spherical nucleic acid (SNA). DKK1 and SFRP1 siRNA treatment were performed in HEK293 cells, human dermal papilla cells (hDPC), and human HFs ex vivo. Functional effects of single and combined DKK1 and SFRP1 knockdown were analyzed in human HFs ex vivo by quantitative (immuno)histomorphology. AR knockdown decreased SFRP1 and DKK1 expression. We found reciprocal mRNA upregulation between DKK1 and SFRP1 following their siRNA knockdown in HEK293 and hDPC. We therefore applied a single and combined treatment of DKK1 and SFRP1 siRNA in HFs ex vivo. SFRP1 knockdown prolonged anagen, increased hair matrix keratinocyte proliferation, reduced apoptosis, and increased DKK1 levels in HFs ex vivo, whereas DKK1 knockdown had no effect, and combined knockdown did not enhance SFRP1’s benefits. The culture-dependent compensatory regulation of SFRP1 and DKK1 underscores Wnt-signaling complexity in hair growth and strengthens the rationale for SFRP1 based therapies in anagen maintenance and hair loss. Full article

Secreted frizzled-related protein 3 (sFRP3/FRZB) is a soluble Wnt antagonist with established roles in skeletal development, however, its specific function in myogenesis remains underexplored. This study investigated the regulatory role of FRZB in muscle development, hypothesizing that it contributes to breed-specific growth differences in pigs. We examined FRZB expression in fetal tissues of slow-growing (Tibetan and Wujin) and fast-growing (Large White) pigs, and assessed its function in C2C12 myoblasts via siRNA-mediated knockdown. FRZB was widely expressed across porcine fetal tissues, with significantly higher abundance in the longissimus dorsi of slow-growing breeds. In vitro, FRZB silencing significantly enhanced myoblast proliferation and migration. Furthermore, knockdown accelerated differentiation and promoted the formation of longer, thicker multinucleated myotubes, accompanied by the upregulation of myogenic (MyoD, MyoG, MyHC) and fusion (β1-integrin, Myomaker) markers. Transcriptional profiling revealed a shift toward hypertrophy (Fst and Nog upregulation) and away from atrophy (Atrogin1 downregulation). These findings identify FRZB as a negative regulator of myogenesis via the Wnt signaling pathway. The elevated expression in indigenous breeds suggests FRZB may impose a molecular constraint on muscle development, highlighting its potential as a candidate gene for regulating carcass traits. Full article

Although the clownfish, Amphiprion ocellaris (A. ocellaris), is a popular ornamental marine fish worldwide, the mechanisms underlying color pattern variation remain unclear. Given that the Platinum-type clownfish, nearly entirely white, has high economic value, understanding the biological mechanism that accounts for the difference between orange and white colors in A. ocellaris is crucial. To investigate these coloration differences, we performed RNA sequencing analysis and identified differentially expressed genes (DEGs) by comparing white and orange skin samples from three A. ocellaris individuals. A total of 76 DEGs were detected, including 56 downregulated and 20 upregulated genes. DEG sequences were annotated using Danio rerio and Stegastus partitus as reference species, selecting the best hit based on the lowest E-value. A protein–protein interaction (PPI) network and Gene Ontology biological process terms were additionally analyzed. Several DEGs previously reported to be associated with pigmentation, including hpdb, cldn11b, sfrp5, slc2a9, slc2a11b, si:ch211-256m1.8, fhl2, rab38, and ttc39b were identified. Based on the functions of these DEGs, it is inferred that leucophores and xanthophores contribute to both white and orange coloration by modulating related genes, including slc2a11b and slc2a9. Additionally, sfrp5, sost, and sp7 genes were identified to interact with each other in the PPI analysis, with sfrp5 and sost being associated with the Wnt signaling pathway, which contributes to melanocyte specification and osteoblast differentiation. Based on these findings, we propose sost and sp7 as candidate genes that might provide insights relevant to extreme white pigmentation phenotypes, such as those observed in Platinum-type clownfish. For a clearer understanding, further studies integrating quantitative genetics and functional analyses are required. Full article

Background/objectives: Progression of pancreatic ductal adenocarcinoma (PDAC) and other carcinomas relies on cancer-associated fibroblasts (CAFs). A subset of CAFs is derived from adipose stromal cells (ASCs) recruited by tumors and the ASC-CAF conversion has been associated with invasiveness and poor prognosis. Methods: To explore the underlying molecular mechanisms, we used a model based on primary ASCs derived from human visceral adipose tissue co-cultured with human PDAC cell line Capan-1. To investigate cancer progression in vivo, we also used mice orthotopically grafted with mouse KPC cells. Results: Genomic analysis revealed that Capan-1 co-culture induces Wnt and TGFβ signaling and extracellular matrix (ECM) gene expression in ASC. We investigated the function of two markers of the fibroblastic transition highly induced by cancer cells: a long non-coding RNA LINC01614 and a Wnt signaling modulator SFRP4. By using ASCs with either SFRP4 or LINC01614 knocked out (ko), we showed that both genes are required for Wnt/TGFβ signaling and ECM induction in ASCs by Capan-1. Analysis of changes in Capan-1 genes that rely on LINC01614 and SFRP4 expression in ASCs also identified the Wnt and TGF pathways. SFRP4 ko in ASCs suppressed both migration and invasion of Capan-1 cells. We show that tumors in SFRP4 ko mice have less desmoplasia, less epithelial dedifferentiation, reduced growth rate, and reduced progression to metastasis. Conclusions: We conclude that SFRP4 promotes cancer progression in pancreatic cancer and is a promising therapeutic target. Full article

Background: Pancreatic adenocarcinoma (PAAD), often referred to as the “king of cancers,” remains poorly understood in terms of the regulatory mechanisms involving brown adipocytes (BAs). Methods: Bioinformatics approaches were employed to explore the role of BAs in PAAD progression, utilizing transcriptomic data from public databases. Prognostic genes were identified through differential expression analysis, univariate Cox regression, and machine learning. A risk model categorizing patients into high- and low-risk groups was developed, accompanied by a nomogram. Functional analysis, immune microenvironment profiling, somatic mutation analysis, and drug sensitivity testing were performed, with further validation via gene localization, immunohistochemistry, and clinical sample analysis. Results: Six prognostic genes (SERPINB5, CALU, TFRC, LY6D, SFRP1, and GBP2) were identified, with the model and nomogram exhibiting robust predictive performance. Notable differences between the high- and low-risk groups were found in immune pathways, cell infiltration, tumor mutational burden, and drug sensitivity (e.g., axitinib). Conclusions: SERPINB5, SFRP1, and TFRC were highly expressed in PAAD samples, providing new insights into potential therapeutic strategies in PAAD treatment. Full article

Background/Objectives: Colorectal cancer (CRC) is a major contributor to cancer-related deaths worldwide. While existing screening tools are effective, their high cost and limited availability restrict widespread adoption, particularly in low- and middle-income settings. The identification of affordable, non-invasive biomarkers is therefore critical to improve early CRC detection and survival outcomes. Methods: A systematic literature search was performed through PubMed, ScienceDirect, Medline, ISI Web of Knowledge, and Google Scholar to identify studies reporting stool- and blood-based biomarkers for CRC detection. Data were extracted using a standardized template, including study details, specimen type, detection method, and diagnostic performance parameters such as sensitivity and specificity. Results: DNA methylation biomarkers demonstrated high diagnostic potential. Syndecan 2 (SDC2) and Short Stature Homeobox 2 (SHOX2) achieved a combined stool sensitivity of 91.35%. Other methylation markers, including NDRG4, SEPT9, and BCAT1, showed a composite sensitivity of 82.7%. Plasma-based methylation markers such as GATA5, FOXE1, and SYNE1 reported sensitivities ranging from 18–47% and specificities of 93–99%. Hypermethylation of SFRP2 and WIF-1 achieved 81.3% sensitivity in CRC and precursor lesions. Matrix metalloproteinases (MMP-2 and MMP-9) were elevated in CRC patients, with stool MMP-9 yielding 72.2% sensitivity and 95% specificity. A stool gene panel (UBE2N, IMPDH1, DYNC1LI1, HRASLS2) reached 96.6% sensitivity and 89.7% specificity, while a methylation-based panel (ALX4, BMP3, NPTX2, RARB, SDC2, SEPT9, VIM) achieved 90.7% sensitivity. MicroRNAs (miR-21, miR-92a, miR-223, miR-182) showed excellent diagnostic performance, with sensitivities exceeding 96% and specificities above 75%. Conclusions: DNA methylation and microRNA biomarkers hold strong promises for non-invasive CRC screening. Multi-marker panels demonstrate superior diagnostic accuracy and may provide a cost-effective, scalable approach for early CRC detection in resource-limited settings. Full article

Dermal papilla cells (DPCs) serve as the signaling hub regulating hair follicle (HF) development and cyclical growth. This study aims to investigate the biological function and molecular mechanisms of TGFBR1 (transforming growth factor β receptor 1), a differentially expressed gene identified through single-cell transcriptomic sequencing (scRNA-seq) in the DPCs from fine-wool sheep. Primary DPCs were isolated and purified using a combination of enzymatic digestion and mechanical dissociation, followed by immunofluorescence identification (α-SMA and SOX2-positive). Following successful transfection with constructed TGFBR1 overexpression plasmids and siRNA interference vectors, cell proliferation was assessed via EDU staining and CCK-8 assays. mRNA expression of key genes in Wnt/β-catenin, BMP, and Notch signaling pathways (PCNA, CCND1, CTNNB1, SFRP2, BMP2, NOTCH3, SMAD4, etc.) was validated by RT-qPCR. Single-cell transcriptomics revealed significant downregulation of TGFBR1 in DPCs from fine-wool sheep. Functional validation demonstrated that TGFBR1 overexpression markedly suppressed DPC proliferation, whereas knockdown of TGFBR1 expression promoted DPC proliferation. Molecular mechanism studies showed that TGFBR1 overexpression significantly downregulated PCNA, CCND1, CTNNB1, NOTCH3, and SMAD4 while upregulating SFRP2, BMP2, and TGFB1 expression. These findings demonstrate that TGFBR1 acts as a negative regulator of DPCs proliferation by modulating the activity of multiple signaling pathways, including Wnt/β-catenin, BMP, and Notch, thereby suppressing the proliferative capacity of DPCs. This study not only provides new theoretical support for elucidating the role of the TGF-β signaling pathway in H development but also offers theoretical reference for in-depth research on molecular breeding in ultra -fine-wool sheep and the molecular mechanisms underlying HF development. Full article

Cardiovascular diseases have become a leading global health burden, with rising mortality worldwide. WNT and JAK/STAT have been highlighted as emerging biomarkers in cardiovascular disease pathogenesis. This study assessed the Wnt/JAK-STAT signaling pathway in relation to SFRP5 and genetic polymorphisms in cardiac patients. This prospective case–control study included 100 patients with various cardiac diseases (IHD, valvular heart disease, HF, cardiomyopathy, and arrhythmia) and 50 matched healthy controls. Clinical and echocardiographic assessments were performed. Plasma SFRP5, Wnt5a, and JAK levels were measured using ELISA; STAT5A expression by flow cytometry; and SFRP5 (rs780369540) gene polymorphism by TaqMan real-time PCR were also performed in all participants. Cardiac patients showed significantly higher median BMI (33 vs. 28.5 kg/m², p = 0.001) and markedly increased median value of each Wnt5a (16.85 vs. 5.6 pg/mL, p < 0.001), median JAK (9.45 vs. 2.4 pg/mL, p < 0.001), and STAT5A expression (87.55% vs. 33%, p < 0.001), with lower SFRP5 levels (4 vs. 6.7 ng/L, p < 0.001) compared to control. The SFRP5 (rs780369540) T allele was more frequent in patients (51.5% vs. 32%, p = 0.001), and dominant TT + TC genotypes were higher (66% vs. 42%, p = 0.005) compared to the control group. TT carriers showed higher median Wnt5a, lower median SFRP5, and reduced ejection fraction compared to other genotypes (TC, CC) carriers. Multivariate analysis identified elevated Wnt5a, JAK, and decreased SFRP5 as independent predictors of cardiovascular disease (p < 0.05). Cardiac patients showed altered WNT5a, JAK, and SFRP5 levels. SFRP5 polymorphism predicted cardiovascular risk independently. Full article

Background: The aim of this study is to investigate the pathophysiology of cataract by analyzing signaling pathways in three sample types obtained from four different lens groups: age-related (ARC), diabetic (DC), post-vitrectomy cataract (PVC) and clear control lenses. Methods: Three sample types—the aqueous humor, the anterior capsule and the phaco cassette content—were collected during cataract surgery from 39 participants (ARC = 12, DC = 11, PVC = 7 and control = 9). The samples were prepared based on Sp3 protocol. The recognition and quantification of proteins were performed with liquid chromatography online with tandem mass spectrometry using the DIA-NN software. Perseus software (v1.6.15.0) was used for statistical analysis. Data are available via ProteomeXchange with identifiers PXD045547, PXD045554, PXD045557, and PXD069667. Results: In total, 1986 proteins were identified in the aqueous humor, 2804 in the anterior capsule, and 3337 in the phaco cassette samples. Proteins involved in actin and microtubule cytoskeleton organization, including ACTN4, were downregulated in all three cataract groups compared to controls. Proteins involved in glycolipid metabolic process, including GAL3ST1, GAL3ST4, and GLA, were upregulated in ARC compared to controls. Proteins involved in the non-canonical Wnt receptor signaling pathway, including FRZB, SFRP1, SFRP2, SFRP5, WNT5A, and WNT7A, were upregulated in ARC compared to DC, PVC, and controls. Conclusions: Comprehensive proteomic profiles were generated using DIA proteomics by comparing ARC, DC, and PVC versus controls. This is the first study to use phaco cassette contents to investigate cataract formation in comparison to controls. Our findings significantly enhance the current understanding of human cataract pathophysiology and provide novel insights into the mechanisms underlying cataract formation. Full article

Short fibre-reinforced polymers (SFRPs) are increasingly used in structural applications where mechanical integrity under complex loading is critical. However, conventional modelling approaches often fail to accurately predict mechanical behaviour in weldline regions formed during injection moulding, where microstructural anomalies and pre-existing damage significantly degrade performance. This study addresses these limitations by extending a hybrid micro–macromechanical constitutive framework to incorporate localised initial damage at weldlines. Calibration and validation of the model were conducted using directional tensile tests on dumbbell-shaped polyamide 66 specimens reinforced with 25 wt% glass fibres, featuring controlled weldline geometry. Digital image correlation (DIC) was employed to capture strain fields, while injection moulding simulations provided fibre orientation distributions and weldline positioning. Results demonstrate that incorporating initial damage and its independent evolution for the cold weld region significantly improves prediction accuracy in weldline zones without compromising model efficiency. The proposed approach can be integrated seamlessly with existing finite element framework and offers a robust solution for simulating SFRP components with weldlines, enhancing reliability in safety-critical applications. Full article

Colorectal cancer (CRC) remains one of the leading causes of cancer-related morbidity and mortality worldwide. Despite significant advances in screening and treatment, the prognosis for advanced-stage disease continues to be poor. One thriving area of research focuses on the use of epigenetic alterations for the diagnosis, prediction of treatment response, and prognosis of CRC. In this study, we evaluated original studies and meta-analyses published within the past five years to identify the most clinically relevant epigenetic biomarkers. DNA methylation-based assays, particularly those targeting SDC2 and SEPT9 in stool and plasma, exhibit superior diagnostic accuracy compared to other epigenetic modalities. Circulating microRNAs (miRNAs), including miR-211, miR-197, and miR-21, as well as specific long non-coding RNAs (lncRNAs) such as SNHG14, LINC01485, and ASB16-AS1, also show promising diagnostic potential. Furthermore, panels combining multiple epigenetic markers, especially those incorporating DNA methylation targets, have demonstrated improved sensitivity and specificity for early-stage CRC detection. In the context of therapeutic prediction, microRNAs such as miR-140, miR-21, and miR-4442 have been associated with chemotherapy resistance and recurrence risk. DNA methylation markers like LINE-1, mSEPT9 and ERCC1 have also shown predictive value, while lncRNAs including MALAT1 and GAS6-AS1 remain less validated. Regarding prognosis, miRNAs appear to be the most promising biomarkers, with miR-675-5p and miR-150 being associated with poor survival, while miR-767-5p and miR-215 predict favorable outcomes. Methylation of NKX6.1, IGFBP3, and LMX1A has been identified as an independent negative prognostic factor, while SFRP2 hypermethylation is linked to better prognosis. Selected lncRNAs, including THOR and LINC01094, have also demonstrated significant prognostic value. Despite these advances, challenges persist, including inconsistent reporting, limited external validation, and a lack of replication by independent research groups. Full article