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We prepared novel plasmon-coupled silver (Ag) and silver oxide (Ag2O) nanoparticles using green synthesis and a magnetic stirrer device from lemon juice. Firstly, we prepared six Ag samples from green tea and lemon juice, characterized by XRD, FTIR, UV, and HRTEM. We selected just two samples to apply to MDA-MB-231 cells. The samples entered the cell through endocytosis, showed moderate cytotoxicity and ROS levels, caused cell growth arrest at the G2M phase, exhibited higher inhibition of Cyclin D1, and induced early apoptosis. β-Catenin is an abundant protein in triple-negative breast cancer TNBC. Both samples showed inhibition of the β-catenin proteins pathway. Plasmon-coupled nanoparticles effectively inhibited β-catenin, physically capturing β-catenin and its pathway proteins, mimicking the action of a degradation protein complex due to their geometric properties. The prepared materials could be considered a promising treatment for TNBC that has not responded to chemotherapy and radiotherapy. As they have low toxicity to normal cells; this paves the way for new material designs without strict size limitations. [ABSTRACT FROM AUTHOR]

Background: Vitiligo is a complex disorder characterized by skin depigmentation; the canonical Wnt signaling pathway that involves β-catenin plays a crucial role in promoting the melanin production in melanocytes. Targeted inhibition of the Janus kinase JAK-STAT pathway can effectively diminish the secretion of the chemokine C-X-C motif ligand CXCL10, thereby safeguarding melanocytes. Ferula has been applied as a treatment regimen for a long period; however, its use for the treatment of vitiligo has not been previously documented. Methods: CCK-8 assay, Intracellular melanin content assay, Tyrosinase activity assay, Western blotting, qRT-PCR, and ELISA methods were employed. Using molecular docking verified the inhibitory effects of feshurin on the JAK1. Results: The sesquiterpene coumarin feshurin was separated from Ferula samarcandica. Feshurin was shown to induce GSK-3β phosphorylation, resulting in the translocation of β-catenin into the nucleus. This translocation subsequently upregulated the transcription of microphthalmia-associated transcription factor (MITF), leading to increased tyrosinase activity and melanin production. In addition, feshurin inhibited the production of chemokine CXCL10 via the JAK-STAT signaling pathway, which was verified by molecular docking. Conclusions: Based on these findings, it can be concluded that feshurin exhibits significant potential for the development of novel anti-vitiligo therapeutics. [ABSTRACT FROM AUTHOR]

Abstract We prepared novel plasmon-coupled silver (Ag) and silver oxide (Ag2O) nanoparticles using green synthesis and a magnetic stirrer device from lemon juice. Firstly, we prepared six Ag samples from green tea and lemon juice, characterized by XRD, FTIR, UV, and HRTEM. We selected just two samples to apply to MDA-MB-231 cells. The samples entered the cell through endocytosis, showed moderate cytotoxicity and ROS levels, caused cell growth arrest at the G2M phase, exhibited higher inhibition of Cyclin D1, and induced early apoptosis. β-Catenin is an abundant protein in triple-negative breast cancer TNBC. Both samples showed inhibition of the β-catenin proteins pathway. Plasmon-coupled nanoparticles effectively inhibited β-catenin, physically capturing β-catenin and its pathway proteins, mimicking the action of a degradation protein complex due to their geometric properties. The prepared materials could be considered a promising treatment for TNBC that has not responded to chemotherapy and radiotherapy. As they have low toxicity to normal cells; this paves the way for new material designs without strict size limitations.

Abstract Background Platinum-based chemotherapy regimens are a mainstay in the management of ovarian cancer (OC), but emergence of chemoresistance poses a significant clinical challenge. The persistence of ovarian cancer stem cells (OCSCs) at the end of primary treatment contributes to disease recurrence. Here, we hypothesized that the extracellular matrix protects CSCs during chemotherapy and supports their tumorigenic functions by activating integrin-linked kinase (ILK), a key enzyme in drug resistance. Methods TCGA datasets and OC models were investigated using an integrated proteomic and gene expression analysis and examined ILK for correlations with chemoresistance pathways and clinical outcomes. Canonical Wnt pathway components, pro-survival signaling, and stemness were examined using OC models. To investigate the role of ILK in the OCSC-phenotype, a novel pharmacological inhibitor of ILK in combination with carboplatin was utilized in vitro and in vivo OC models. Results In response to increased fibronectin secretion and integrin β1 clustering, aberrant ILK activation supported the OCSC phenotype, contributing to OC spheroid proliferation and reduced response to platinum treatment. Complexes formed by ILK with the Wnt receptor frizzled 7 (Fzd7) were detected in tumors and correlated with metastatic progression. Moreover, TCGA datasets confirmed that combined expression of ILK and Fzd7 in high grade serous ovarian tumors is correlated with reduced response to chemotherapy and poor patient outcomes. Mechanistically, interaction of ILK with Fzd7 increased the response to Wnt ligands, thereby amplifying the stemness-associated Wnt/β-catenin signaling. Notably, preclinical studies showed that the novel ILK inhibitor compound 22 (cpd-22) alone disrupted ILK interaction with Fzd7 and CSC proliferation as spheroids. Furthermore, when combined with carboplatin, this disruption led to sustained AKT inhibition, apoptotic damage in OCSCs and reduced tumorigenicity in mice. Conclusions This “outside-in” signaling mechanism is potentially actionable, and combined targeting of ILK-Fzd7 may lead to new therapeutic approaches to eradicate OCSCs and improve patient outcomes.

Sensorineural hearing loss can be caused by lesions to the inner ear during development. Understanding the events and signaling pathways that drive inner ear formation is crucial for determining the possible causes of congenital hearing loss. We have analyzed the innervation and expression of SOX2, JAGGED1, β-catenin (CTNNB1), and vitamin D receptor (VDR) in the inner ears of human conceptuses aged 5 to 10 weeks after fertilization (W) using immunohistochemistry. The prosensory domains of the human inner ear displayed SOX2 and JAGGED1 expression throughout the analyzed period, with SOX2 expression being more extensive in all the analyzed timepoints. Innervation of vestibular prosensory domains was present at 6 W and extensive at 10 W, while nerve fibers reached the base of the cochlear prosensory domain at 7–8 W. CTNNB1 and VDR expression was mostly membranous and present during all analyzed timepoints in the inner ear, being the strongest in the non-sensory epithelium. Their expression was stronger in the vestibular region compared to the cochlear duct. CTNNB1 and VDR expression displayed opposite expression trends during the analyzed period, but additional studies are needed to elucidate whether they interact during inner ear development. [ABSTRACT FROM AUTHOR]

Background: Colorectal cancer ranks third among the most common cancer of male. Globally colorectal adenocarcinoma accounts for approximately 1.2 million new cases, and 600,000 deaths each year1. In India, the incidence of colorectal cancer is 4.3 and 3.4 per 100,000 male and female respectively2. Colorectal carcinoma (CRC) is considered as the most preventable cancers because it arises from benign neoplasms, called polyps which evolve into colorectal carcinomas over many years. The aim of the study is to study the age, sex, site and clinical presentation of various colorectal neoplasm, to study expression of beta catenin in benign, premalignant and malignant tumors of colorectum and correlate the level of expression of beta catenin in various histological grades of colorectal carcinoma. Materials and Methods: Paraffin blocks from 23 cases has been selected which includes both polyps and colorectal carcinoma. Colorectal carcinoma has been graded and staged according to the cap protocol. Polyps has been classified in to benign and premalignant, whereas colorectal carcinoma has been graded with respect to degree of differentiation. Betacatenin immuno-histochemistry has been done. Scoring has done with respect to the localization of beta-catenin in membrane, cytoplasm and nucleus. Result: Normal colorectal mucosa and benign polyps shows predominant membranous expression. Premalignant polyp shows predominantly increased cytoplasmic expression with focal nuclear positivity and the colorectal carcinoma shows raised nuclear expression and loss of membranous expression. This proves the gradual translocation of beta catenin from membrane to nucleus in adenomacarcinoma sequence. Beta-catenin expression has been significantly correlated with depth of invasion (T), nodal metastasis (N) and thereby to the stage of the carcinoma. Beta-catenin expression has been statistically correlated with degree of differentiation (tumor grade). Both recurrence and death were equally distributed in our study, through this observation the morbidity and mortality were greatly correlated with grade of beta-catenin expression. Hence beta-catenin can be used as a marker to identify the malignant potential of premalignant dysplastic polyps which helps in early management. It is used to assess the prognosis of the patients who were in post-colectomy status hence this marker can be used as an additional prognostic marker. [ABSTRACT FROM AUTHOR]

The blood-brain barrier consists of tightly connected endothelial cells protecting the brain's microenvironment from the periphery. These endothelial cells are characterized by specific tight junction proteins such as Claudin-5 and Occludin, forming the endothelial barrier. Disrupting these cells might lead to blood-brain barrier dysfunction. The Wnt/β-catenin signaling pathway can regulate the expression of these tight junction proteins and subsequent barrier permeability. The aim of this study was to investigate the in vitro effects of Wnt7a mediated β-catenin signaling on endothelial barrier integrity. Mouse brain endothelial cells, bEnd.3, were treated with recombinant Wnt7a protein or XAV939, a selective inhibitor of Wnt/β-catenin mediated transcription to modulate the Wnt signaling pathway. The involvement of Wnt/HIF1α signaling was investigated by inhibiting Hif1α signaling with Hif1α siRNA. Wnt7a stimulation led to activation and nuclear translocation of β-catenin, which was inhibited by XAV939. Wnt7a stimulation decreased Claudin-5 expression mediated by β-catenin and decreased endothelial barrier formation. Wnt7a increased Hif1α and Vegfa expression mediated by β-catenin. However, Hif1α signaling pathway did not regulate tight junction proteins Claudin-5 and Occludin. Our data suggest that Wnt7a stimulation leads to a decrease in tight junction proteins mediated by the nuclear translocation of β-catenin, which hampers proper endothelial barrier formation. This process might be crucial in initiating endothelial cell proliferation and angiogenesis. Although HIF1α did not modulate the expression of tight junction proteins, it might play a role in brain angiogenesis and underlie pathogenic mechanisms in Wnt/HIF1α signaling in diseases such as cerebral small vessel disease. [ABSTRACT FROM AUTHOR]

BackgroundEmpagliflozin (EMPA) is an SGLT-2 inhibitor that can control hyperglycemia. Clinical trials have indicated its cardio-protective effects against cardiac remodeling in diabetes or non-diabetes patients. However, the underlying molecular mechanisms of EMPA’s cardio-protective effects remain elusive.MethodsWe evaluated whether the EMPA attenuated the pressure-overload-induced cardiac hypertrophy by inhibiting the Wnt/β-catenin pathway. Furthermore, the effects of the EMPA on a mouse model of transverse aortic constriction (TAC) induced cardiac hypertrophy was also evaluated. Mice were administrated with 0.5% CMC-Na as a vehicle or EMPA (10 mg/kg/day, daily, throughout the study) by intragastric gavage.ResultsThe in vivo echocardiography and histologic morphological analyses revealed that EMPA attenuated TAC-induced cardiac hypertrophy. Moreover, it also ameliorated TAC-induced cardiac fibrosis and decreased the cell size of the cardiomyocytes in isolated adult cardiomyocytes. Molecular mechanism analysis revealed that the EMPA reduced the TAC-induced enhanced expression of the Wnt/β-catenin pathway in vivo. For in vitro assessments, isolated neonatal rat cardiomyocytes (NRCMs) were treated with Angiotensin II (AngII) and EMPA; the results showed that in the absence of EMPA, the expression of the Wnt/β-catenin pathway was enhanced. In the trans-genetic heterozygous β-catenin deletion mice, EMPA attenuated TAC-induced cardiac remodeling by reducing the Wnt/β-catenin pathway. In addition, molecular docking analysis indicated that EMPA interacts with FZD4 to inhibit the TAC and AngII induced Wnt/β-catenin pathway in cardiomyocytes.ConclusionOur study illustrated that EMPA might directly interact with FZD4 to inhibit the TAC and AngII-induced activation of the Wnt/β-catenin pathway to attenuate the adverse cardiac remodeling.

Pulmonary blastomas (PB) are rare, aggressive tumours which constitute

The epithelial-mesenchymal transition (EMT) plays a crucial role in the development of pancreatic ductal adenocarcinoma (PDAC). There are insufficient and contradictory data in the scientific literature on the peculiarities of epithelial and mesenchymal marker expression in PDAC ducts and EMT zone, which requires further research on the role of EMT in the development and progression of PDAC. Aim: to conduct a comprehensive assessment of epithelial and mesenchymal markers of EMT in the PDAC ducts and EMT zone at different degrees of differentiation. Materials and methods. A comprehensive pathomorphological and immunohistochemical examination including 49 cases of surgical material from patients with PDAC, divided into groups of moderate (G2) and low degree of tumor differentiation (G3). Results. PDAC was characterized by low levels of E-cadherin expression in both ducts and EMT – Me = 22.58 % [12.81; 36.23] and Me = 25.17 % [19.04; 35.37], respectively (p > 0.05). Only membrane expression of the marker was detected in the ducts and membrane-cytoplasmic one – in the EMT zone without significant differences in the groups. There was a significant decrease in the ductal β-catenin expression (p < 0.05) in G2 with membrane-cytoplasmic staining (Me = 15.58 % [10.42; 26.24]), in G3 – with membrane (Me = 4.42 % [2.35; 5.93]); in the EMT zone, only membrane-cytoplasmic expression was observed in both groups without significant difference (p > 0.05). Membrane expression of CK7 was positive in 100 % of PDAC, significantly lower at G2 (Me = 19.51 % [10.70; 27.24]; Me = 26.19 % [20.93; 30.05], p < 0.05), and CK18 (Me = 21.34 % [9.68; 29.96] – in G2, in G3 – Me = 22.50 % [8.24; 40.08], p > 0.05). Expression of α-SMA and Vim was seen in spindle-shaped stromal cells, around tubular and trabecular structures with membrane-cytoplasmic staining. There was no significant difference between α-SMA in G2 and G3, the level of vimentin was significantly lower in G3 (p < 0.05). Conclusions. The features of PDAC E-cadherin, β-catenin, CK7 and CK18 marker expression indicate a greater EMT process at the periphery of the tumor and its severity increases with tumor progression. The optimal markers to determine the mesenchymal phenotype of PDAC cells are α-SMA and vimentin.

Activation of the Wnt pathway lies at the core of many human cancers. Wnt and macropinocytosis are often active in the same processes, and understanding how Wnt signaling and membrane trafficking cooperate should improve our understanding of embryonic development and cancer. Here, we show that a macropinocytosis activator, the tumor promoter phorbol 12-myristate 13-acetate (PMA), enhances Wnt signaling. Experiments using the Xenopus embryo as an in vivo model showed marked cooperation between the PMA phorbol ester and Wnt signaling, which was blocked by inhibitors of macropinocytosis, Rac1 activity, and lysosome acidification. Human colorectal cancer tissue arrays and xenografts in mice showed a correlation of cancer progression with increased macropinocytosis/multivesicular body/lysosome markers and decreased GSK3 levels. The crosstalk between canonical Wnt, focal adhesions, lysosomes, and macropinocytosis suggests possible therapeutic targets for cancer progression in Wnt-driven cancers.

BackgroundNeural stem cells (NSCs) are considered as candidates for cell replacement therapy in many neurological disorders. However, the propensity for their differentiation to proceed more glial rather than neuronal phenotypes in pathological conditions limits positive outcomes of reparative transplantation. Exogenous physical stimulation to favor the neuronal differentiation of NSCs without extra chemical side effect could alleviate the problem, providing a safe and highly efficient cell therapy to accelerate neurological recovery following neuronal injuries.ResultsWith 7-day physiological electric field (EF) stimulation at 100 mV/mm, we recorded the boosted neuronal differentiation of NSCs, comparing to the non-EF treated cells with 2.3-fold higher MAP2 positive cell ratio, 1.6-fold longer neuronal process and 2.4-fold higher cells ratio with neuronal spontaneous action potential. While with the classical medium induction, the neuronal spontaneous potential may only achieve after 21-day induction. Deficiency of either PI3Kγ or β-catenin abolished the above improvement, demonstrating the requirement of the PI3K/Akt/GSK-3β/β-catenin cascade activation in the physiological EF stimulation boosted neuronal differentiation of NSCs. When transplanted into the spinal cord injury (SCI) modelled mice, these EF pre-stimulated NSCs were recorded to develop twofold higher proportion of neurons, comparing to the non-EF treated NSCs. Along with the boosted neuronal differentiation following transplantation, we also recorded the improved neurogenesis in the impacted spinal cord and the significantly benefitted hind limp motor function repair of the SCI mice.ConclusionsIn conclusion, we demonstrated physiological EF stimulation as an efficient method to boost the neuronal differentiation of NSCs via the PI3K/Akt/GSK-3β/β-catenin activation. Pre-treatment with the EF stimulation induction before NSCs transplantation would notably improve the therapeutic outcome for neurogenesis and neurofunction recovery of SCI.

Sinonasal glomangiopericytoma is an uncommon mesenchymal tumor with a perivascular myoid phenotype, which is categorized as a borderline/low-grade malignant soft tissue tumor by the current World Health Organization Classification of Head and Neck tumors. Here, we present the case of a 53-year-old woman with an unusual spindle cell morphology of sinonasal glomangiopericytoma arising in the nasal cavity, mimicking solitary fibrous tumor. Microscopically, the tumor showed a cellular proliferation of spindle cells in fascicles including a focal long sweeping arrangement or whorls, or with a storiform growth pattern, associated with hemangiopericytoma-like gaping blood vessels embedded in a fibrous stroma. This arrangement of the spindle cells faintly indicated a solitary fibrous tumor rather than sinonasal glomangiopericytoma. Immunohistochemically, the tumor was positively reactive to not only beta-catenin (in the nuclei) but also CD34, although signal transducers and activators of transcription 6 was negative. Mutational analysis using Sanger sequencing detected a CTNNB1 mutation. We finally diagnosed the tumor as a sinonasal glomangiopericytoma, showing an unusual spindle cell variant. Such unusual spindle cell morphology with CD34-immunoreactivity potentially leads to an incorrect diagnosis of solitary fibrous tumor because such prominent fascicles including long sweeping structures, reminiscent of desmoid-type fibromatosis, have scarcely been described in the literature. Hence, careful morphological scrutiny using appropriate diagnostic adjuncts is necessary for correct diagnosis. [ABSTRACT FROM AUTHOR]

Desmoid tumors are locally aggressive, benign neoplasms originating in connective tissues. Although the exact pathophysiology remains unknown, antecedent trauma or surgery are believed to be important contributing factors. The occurrence of paraspinal desmoid tumor in pediatric patients is extremely uncommon. Here, we present an exceedingly rare case of a pediatric patient with no surgical or family history who developed a paraspinal desmoid tumor. A 9-year-old female patient presented with 4 months of progressive back pain, right lower extremity weakness, and numbness. Spinal imaging revealed a left epidural paraspinal mass compressing her thoracic spinal cord and extending into the left thoracic cavity. A multidisciplinary approach with neurosurgery and thoracic surgery enabled gross total resection of the lesion. The patient had complete resolution of her symptoms with no signs of residual tumor on postoperative imaging. Pathology revealed a desmoid tumor that avidly stained for beta-catenin. On her last followup, she developed a recurrence, to which she was started on sorafenib therapy. Desmoid tumors are rare connective tissue neoplasms that often occur after local tissue trauma, such as that caused by surgery. This report presents a rare case of a pediatric paraspinal desmoid tumor that occurred in a patient with no surgical or family history. Such tumors should undergo surgical resection for symptomatic relief and tissue diagnosis. Close clinical and radiographic surveillance are essential in these patients due to the high recurrence rates of desmoid tumor. [ABSTRACT FROM AUTHOR]

Background: A myriad of therapeutic modalities for alopecia areata are available; however, none is of high level of evidence, creating an immense need for the evaluation of other treatment modalities, of which topical sodium valproate is of potential role via proposed decrease in beta-catenin breakdown, despite its well-known side effect of hair fall as an oral therapy. Objective: Evaluating the efficacy and the safety of sodium valproate (SV)-loaded nanospanlastics, in comparison to topical corticosteroids, this is the currently available gold standard topical treatment for patchy AA. Methodology: A total of 66 patients with patchy AA were randomly assigned to receive either topical mometasone furoate lotion or topical SV applied twice daily to all patches except a control patch, which was left untreated. Clinical, trichoscopic and biochemical assessments of beta-catenin tissue levels and Axin-2 gene expression were carried out at baseline and after 3 months. Results: Both therapeutic modalities were comparable. Potential efficacy was highlighted by significant improvement in the representative patch, the largest treated patch, to the control patch, the smallest untreated patch in both steroid and valproate groups (p = 0.027, 0.003 respectively). Both beta-catenin levels and Axin-2 gene expression were reduced after treatment, pointing to the inhibitory effect of dominating uncontrolled inflammatory milieu. Baseline beta-catenin was found to significantly negatively correlate with improvement in the representative patch in patients with baseline level above 0.42 ng/ml (p = − 0.042). Conclusion: Both topical SV and steroids are of comparable modest efficacy. Thus, further evaluation of SV is due in combination with intralesional steroids and other anti-inflammatory treatment modalities, together with developing individualized approaches based on baseline beta-catenin level. ClinicalTrials.gov Identifier: NCT05017454, . [ABSTRACT FROM AUTHOR]

Background: Hair loss occurs due to various biological and environmental causes, which can have psychosocial consequences. The Wnt/β‐catenin signaling is well‐known for its role in hair growth and regeneration, as it induces the proliferation and differentiation of hair cells. When the leucine‐rich G protein‐coupled receptor 5 (Lgr5) interacts with the R‐spondins, the frizzled receptor (FZD), a Wnt receptor, becomes stabilized, resulting in an increased β‐catenin activity. Aim: We investigated whether the octapeptide that binds to Lgr5 enhances proliferation and differentiation of human primary hair cells through the activation of Wnt/β‐catenin signaling. Methods: The binding affinity of the octapeptide to Lgr5 was evaluated using surface plasmon resonance (SPR). We confirmed changes in proliferation and related factors like β‐catenin activation and growth factors (GFs) expression in human hair follicle dermal papilla cells (HHFDPCs). Additionally, we observed the proliferation and the expression of differentiation markers in human hair follicle outer root sheath cells (HHFORSCs), human hair follicle germinal matrix cells (HHFGMCs), and human hair follicle stem cells (HHFSCs). We used three‐dimensional HHFDPC spheroid culture treated with dihydrotestosterone (DHT) to create in vitro conditions that mimic androgenetic alopecia, and we studied the effects of octapeptide on Wnt expression and HHFSC differentiation. Results: The binding of the octapeptide to Lgr5 was confirmed using SPR analysis. In HHFDPCs, treatment with octapeptide resulted in a concentration‐dependent increase in proliferation. We also observed increased nuclear translocation of β‐catenin and increased expression of its downstream targets. HHFDPCs treated with octapeptide exhibited increased expression of growth factors and phosphorylation of Akt and ERK. In addition, we confirmed that octapeptide increased proliferation and induced differentiation in HHFORSCs, HHFGMCs, and HHFSCs. Under the HHFDPC spheroid culture conditions, we found that octapeptide restored the inhibition of Wnt‐5a and Wnt‐10b expressions by DHT. In HHFSCs treated with HHFDPC spheroid culture media, we observed that octapeptide recovered the inhibition of differentiation by DHT. Conclusion: We found that octapeptides activated the Wnt/β‐catenin signaling and induced the proliferation and differentiation of human primary hair cells by acting as an exogenous ligand for Lgr5. In addition, octapeptides recovered inhibited hair regeneration characters by DHT in androgenetic alopecia‐mimic in vitro model. These findings suggest that octapeptides may be a promising therapeutic option for treating hair loss. [ABSTRACT FROM AUTHOR]

Background: Acute myeloid leukaemia(AML) is a disease of the haematopoietic stem cells. Beta-catenin, a key element of the Wnt signalling, which is particularly active in various cancers. Our aim of this study was to determine beta-catenin gene expression levels in AML patients. Then, it is planned to compare the genes between AML grouped according to beta-catenin gene expression levels by DNA microarray. When the gene group identified in this way is combined with the genes in the control group, the aim is to determine the genes that are associated with beta-catenin in AML. Methods: In this study, beta-catenin gene expression levels were determined in 19 AML patients and 3 controls by qRT-PCR. Transcriptome analysis was performed on AML grouped according to beta-catenin expression levels. Differentially expressed genes(DEGs) were identified and investigated with using DAVID, GO, KEGG and STRING. Results: The transcriptome profiles of our AML samples showed different molecular signature profiles according on their beta-catenin levels(high-low). A total of 20 genes have been identified as hub genes. Among these, TTK, HJURP, KIF14 and BTF3, RPL17, RSL1D1 were found to be associated with poor survival and suggested to be associated with beta-catenin in AML. Furthermore, for the first time in our study, the ELOV6 gene, which is the most highly up-regulated gene in human AML samples, was correlated with a poor prognosis via beta-catenin high levels. Conclusion: It is suggested that the identification of beta-catenin-related gene profiles in AML may help to select new therapeutic targets for the treatment of AML.

CTNNB1, encoding β-catenin protein, is the most frequently altered proto-oncogene in hepatic neoplasms. In this study, we studied the significance and pathological mechanism of CTNNB1 gain-of-function mutations in hepatocarcinogenesis. Activated β-catenin not only triggered hepatic tumorigenesis but also exacerbated Tp53 deletion or hepatitis B virus infection-mediated liver cancer development in mouse models. Using untargeted metabolomic profiling, we identified boosted de novo pyrimidine synthesis as the major metabolic aberration in β-catenin mutant cell lines and livers. Oncogenic β-catenin transcriptionally stimulated AKT2, which then phosphorylated the rate-limiting de novo pyrimidine synthesis enzyme CAD (carbamoyl-phosphate synthetase 2, aspartate transcarbamoylase, dihydroorotase) on S1406 and S1859 to potentiate nucleotide synthesis. Moreover, inhibition of β-catenin/AKT2-stimulated pyrimidine synthesis axis preferentially repressed β-catenin mutant cell proliferation and tumor formation. Therefore, β-catenin active mutations are oncogenic in various preclinical liver cancer models. Stimulation of β-catenin/AKT2/CAD signaling cascade on pyrimidine synthesis is an essential and druggable vulnerability for β-catenin mutant liver cancer.

Background: Colorectal cancer (CRC) has consistently shown a global rise in incidence. p53 exerts regulatory function in signaling processes, with the p53 mutation accelerating the late stage of colorectal carcinogenesis. Programmed death ligand 1 (PD-L1) is an immune checkpoint regulator whose role in targeted therapy of CRCs needs to be evaluated. Beta-catenin plays a fundamental role in cell adhesion, with long-term effects on disease progression and survival. A detailed evaluation of all these factors is essential to tailoring new therapeutic approaches in advanced-stage cases. Aims and Objectives: To study the immunohistochemical (IHC) expression of p53, PD-L1, and beta-catenin in different histologic grades and stages of colorectal carcinomas. Materials and Methods: A total of 29 cases of segmental colectomy were included in the study population. Tumor sections were sent for IHC study of p53, PD-L1, and beta-catenin and interpreted as per standard protocol. Findings were tabulated against the histologic grade and stages of CRCs. Statistical analysis was done using SPSS 25.0 to test for statistical significance at P

Background: Vitiligo is a complex disorder characterized by skin depigmentation; the canonical Wnt signaling pathway that involves β-catenin plays a crucial role in promoting the melanin production in melanocytes. Targeted inhibition of the Janus kinase JAK-STAT pathway can effectively diminish the secretion of the chemokine C-X-C motif ligand CXCL10, thereby safeguarding melanocytes. Ferula has been applied as a treatment regimen for a long period; however, its use for the treatment of vitiligo has not been previously documented. Methods: CCK-8 assay, Intracellular melanin content assay, Tyrosinase activity assay, Western blotting, qRT-PCR, and ELISA methods were employed. Using molecular docking verified the inhibitory effects of feshurin on the JAK1. Results: The sesquiterpene coumarin feshurin was separated from Ferula samarcandica. Feshurin was shown to induce GSK-3β phosphorylation, resulting in the translocation of β-catenin into the nucleus. This translocation subsequently upregulated the transcription of microphthalmia-associated transcription factor (MITF), leading to increased tyrosinase activity and melanin production. In addition, feshurin inhibited the production of chemokine CXCL10 via the JAK-STAT signaling pathway, which was verified by molecular docking. Conclusions: Based on these findings, it can be concluded that feshurin exhibits significant potential for the development of novel anti-vitiligo therapeutics.

With self‐renewal and pluripotency features, embryonic stem cells (ESCs) provide an invaluable tool to investigate early cell fate decisions. Pluripotency exit and lineage commitment depend on precise regulation of gene expression that requires coordination between transcription (TF) and chromatin factors in response to various signaling pathways. SET domain‐containing 3 (SETD3) is a methyltransferase that can modify histones in the nucleus and actin in the cytoplasm. Through an shRNA screen, we previously identified SETD3 as an important factor in the meso/endodermal lineage commitment of mouse ESCs (mESC). In this study, we identified SETD3‐dependent transcriptomic changes during endoderm differentiation of mESCs using time‐course RNA‐seq analysis. We found that SETD3 is involved in the timely activation of the endoderm‐related gene network. The canonical Wnt signaling pathway was one of the markedly altered signaling pathways in the absence of SETD3. The assessment of Wnt transcriptional activity revealed a significant reduction in Setd3‐deleted (setd3∆) mESCs coincident with a decrease in the nuclear pool of the key TF β‐catenin level, though no change was observed in its mRNA or total protein level. Furthermore, a proximity ligation assay (PLA) found an interaction between SETD3 and β‐catenin. We were able to rescue the differentiation defect by stably re‐expressing SETD3 or activating the canonical Wnt signaling pathway by changing mESC culture conditions. Our results suggest that alterations in the canonical Wnt pathway activity and subcellular localization of β‐catenin might contribute to the endoderm differentiation defect of setd3∆ mESCs. [ABSTRACT FROM AUTHOR]

Ameloblastic carcinoma is a rare malignant neoplasm with characteristic histopathological features that are directed towards an aggressive surgical approach than benign odontogenic lesions. It affects people of all ages, mostly in the posterior mandible, without a preference for race or gender. De novo cancer is one of its primary types, while the second type is defined as a malignant change from an antecedent case of benign ameloblastoma. The rapid progression of molecular biology led to the revelation that ameloblastoma contains a BRAF-V600E genetic mutation over 60%. Besides conventional ameloblastic carcinomas, rare histologic variants have also been described in the literature, including clear and spindle cells. These variants pose diagnostic challenges as to whether it is a dedifferentiation or a distinct entity. The dearth of data lends credence to the notion that these histologic variations are related to high-grade neoplasms and more aggressive outcomes. As a result, the current report intends to analyze a series of patients diagnosed with conventional ameloblastic carcinoma of the head and neck region with spindle and clear cell types along with a brief assessment of the literature. [ABSTRACT FROM AUTHOR]

Background: Colorectal cancer (CRC) has consistently shown a global rise in incidence. p53 exerts regulatory function in signaling processes, with the p53 mutation accelerating the late stage of colorectal carcinogenesis. Programmed death ligand 1 (PD-L1) is an immune checkpoint regulator whose role in targeted therapy of CRCs needs to be evaluated. Beta-catenin plays a fundamental role in cell adhesion, with long-term effects on disease progression and survival. A detailed evaluation of all these factors is essential to tailoring new therapeutic approaches in advanced-stage cases. Aims and Objectives: To study the immunohistochemical (IHC) expression of p53, PD-L1, and beta-catenin in different histologic grades and stages of colorectal carcinomas. Materials and Methods: A total of 29 cases of segmental colectomy were included in the study population. Tumor sections were sent for IHC study of p53, PD-L1, and beta-catenin and interpreted as per standard protocol. Findings were tabulated against the histologic grade and stages of CRCs. Statistical analysis was done using SPSS 25.0 to test for statistical significance at P<0.05. Results: The study population predominantly consisted of males (66%), mainly in the age group of 46-60 years. p53 was significantly more expressed in left colonic tumors. PD-L1 expression was also significantly associated with histologic grade and nodal metastasis. High beta-catenin expression was seen in well-differentiated adenocarcinomas and lower T and N-stage disease. Positive p53 also correlated with positive PD-L1 expression. However, expression of p53 was also high in tumors with low beta-catenin expression. Conclusion: p53, PD-L1, and beta-catenin may have a role in prognostication and predicting long-term survival outcomes in CRC cases. These may also act as potential therapeutic targets with a more detailed evaluation and a larger sample size of CRC cases. [ABSTRACT FROM AUTHOR]

Objective: To investigate whether beta-catenin and Forkhead box protein P1 (FOXP1) play a role in pathogenesis of polypoid endometriosis (PE). Material and Method: Our study included fifteen cases of PE. Clinical findings were gathered from archived files of relevant clinics and pathology reports. All glass slides were re-examined for confirmation of the diagnosis and the detection of additional microscopic findings. An immunohistochemical examination was performed using anti beta-catenin and FOXP1 antibodies in fifteen cases of PE, and in a control group that contained nine cases of endometrial polyps (EP) and nine cases of conventional ovarian endometriosis (OE). Result: Stromal nuclear beta-catenin expression was observed in six cases in PE, five cases in EP and one case in the OE group. Stromal FOXP1 staining in PE and EP was significantly reduced as compared to OE. Five PE and two EP cases showed stromal FOXP1 staining while all the OE cases showed stromal FOXP1 staining. The Stromal FOXP1 staining was statistically significant between PE vs OE (p=0.002) and EP vs OE (p=0.023) cases. There was no difference between PE and the control cases in terms of nuclear beta-catenin staining (p=0.69). There was no correlation between these two antibodies and histologic features. Conclusion: The loss of stromal FOXP1 is another biological difference of PE and the overall similarity of expression of FOXP1 between PE and EP could be regarded as a contributing factor for polyp formation. [ABSTRACT FROM AUTHOR]

Simple Summary: Personalised cancer treatment improves outcome for patients but requires that each individual tumour is classified using molecular tools. Extensive genomic studies for each individual patients are out of reach for the majority of the population, therefore, pathologists need to apply readily available tests to achieve molecular classification for all cancer patients. This paper describes how gastric cancer can be classified using on slide tests already used by histopathologists. Using this classification, oncologists can select more effective treatment for each patient. Background and Objectives: Gastric cancer (GC) is one of the most commonly diagnosed cancers and the fourth cause of cancer death worldwide. Personalised treatment improves GC outcomes. A molecular classification is needed to choose the appropriate therapy. A classification that uses on-slide biomarkers and formalin-fixed and paraffin-embedded (FFPE) tissue is preferable to comprehensive genomic analysis. In 2016, Setia and colleagues proposed an on-slide classification; however, this is not in widespread use. We propose a modification of this classification that has six subgroups: GC associated with Epstein–Barr virus (GC EBV+), GC with mismatch-repair deficiency (GC dMMR), GC with epithelial–mesenchymal transformation (GC EMT), GC with chromosomal instability (GC CIN), CG that is genomically stable (GC GS) and GC not otherwise specified (GC NOS). This classification also has a provision for biomarkers for current or emerging targeted therapies (Her2, PD-L1 and Claudin18.2). Here, we assess the implementation and feasibility of this inclusive working classification. Materials and Methods: We constructed a tissue microarray library from a cohort of 79 resection cases from FFPE tissue archives. We used a restricted panel of on-slide markers (EBER, MMR, E-cadherin, beta-catenin and p53), defined their interpretation algorithms and assigned each case to a specific molecular subtype. Results: GC EBV(+) cases were 6%, GC dMMR cases were 20%, GC EMT cases were 14%, GC CIN cases were 23%, GC GS cases were 29%, and GC NOS cases were 8%. Conclusions: This working classification uses markers that are widely available in histopathology and are easy to interpret. A diagnostic subgroup is obtained for 92% of the cases. The proportion of cases in each subgroup is in keeping with other published series. Widescale implementation appears feasible. A study using endoscopic biopsies is warranted. [ABSTRACT FROM AUTHOR]

Purpose: Choledochal cysts are congenital anomalies that occur as localized cystic or fusiform dilatations of the biliary tree. Reflux and stasis of pancreatic enzymes in the biliary duct may relate to the development of intestinal metaplasia which might be an important factor related to the carcinogenesis of choledochal cyst, thus the expression of beta-catenin in the metaplastic epithelium might be associated with malignant transformation of choledochal cyst epithelium. Methods: This study was conducted at a tertiary care pediatric center between October 2014 and March 2017. Forty patients were evaluated for epithelial lining, mural ulceration, fibrosis, inflammation, and metaplasia. Results: Out of 40, 12 cases (30.0%) were the infantile age group and 28 cases (70.0%) were in the classic pediatric group. Ulceration was classified as grade 0 (14 cases, 35.0%), grade 1 (17 cases, 42.5%), or grade 2 (nine cases, 22.5%). Inflammation was classified as grade 0 (2 cases, 5.0%), grade 1 (26 cases, 65.0%), or grade 2 (12 cases, 30.0%). Fibrosis was classified as grade 0 (five cases, 12.5%), grade 1 (11 cases, 27.5%), grade 2 (17 cases, 42.5%), or grade 3 (seven cases, 17.5%). Metaplasia was noted in five (12.5%) out of 40 cases. All choledochal cysts with metaplasia showed beta-catenin nuclear positivity on immunohistochemistry and were followed up. Conclusion: This study emphasizes the importance of detailed histopathological examination and documentation of metaplastic changes. Metaplasia was associated with beta-catenin nuclear positivity. These findings suggest a potential role for beta-catenin as a marker of metaplastic changes in choledochal cysts. [ABSTRACT FROM AUTHOR]

Desmoid fibromatosis is a myofibroblastic neoplasm of intermediate biologic potential, which has a strong predilection for local recurrence but does not metastasize. Arranged in long, sweeping fascicles with infiltrative borders, desmoid fibromatosis typically consists of uniform, bland myofibroblastic spindle cells that harbor mutation of CTNNB1 (or less often APC). In this report, we present a remarkable case of desmoid fibromatosis associated with striking nuclear pleomorphism. We hypothesize that this striking pleomorphism is due to a germline TP53 mutation, a finding first suspected due to strong and diffuse p53 staining by immunohistochemistry and subsequently confirmed by molecular testing. The combination of the pleomorphism and TP53 mutation in desmoid fibromatosis represents a novel finding unreported in the literature. [ABSTRACT FROM AUTHOR]

Mutations in the gene for β-catenin cause liver cancer cells to release fewer exosomes, which reduces the number of immune cells infiltrating the tumor.

Summary: The reprogramming of glutamine metabolism is a key event in cancer more generally and in hepatocellular carcinoma (HCC) in particular. Glutamine consumption supplies tumours with ATP and metabolites through anaplerosis of the tricarboxylic acid cycle, while glutamine production can be enhanced by the overexpression of glutamine synthetase. In HCC, increased glutamine production is driven by activating mutations in the CTNNB1 gene encoding β-catenin. Increased glutamine synthesis or utilisation impacts tumour epigenetics, oxidative stress, autophagy, immunity and associated pathways, such as the mTOR (mammalian target of rapamycin) pathway. In this review, we will discuss studies which emphasise the pro-tumoral or tumour-suppressive effect of glutamine overproduction. It is clear that more comprehensive studies are needed as a foundation from which to develop suitable therapies targeting glutamine metabolic pathways, depending on the predicted pro- or anti-tumour role of dysregulated glutamine metabolism in distinct genetic contexts.

Abstract Background Hematogenous tumor spread of malignant meningiomas occurs very rarely but is associated with very poor prognosis. Case presentation We report an unusual case of a patient with a malignant meningioma who developed multiple metastases in bones, lungs and liver after initial complete resection of the primary tumor. After partial hepatic resection, specimens were histologically analyzed, and a complete loss of E-cadherin adhesion molecules was found. No oncogenic target mutations were found. The patient received a combination of conventional radiotherapy and peptide receptor radionuclide therapy (PRRT). Due to aggressive tumor behavior and rapid spread of metastases, the patient deceased after initiation of treatment. Conclusions E-cadherin downregulation is associated with a higher probability of tumor invasion and distant metastasis formation in malignant meningioma. Up to now, the efficacy of systemic therapy, including PRRT, is very limited in malignant meningioma patients.

Basal‐like breast cancer (BBC) and glioblastoma multiforme (GBM) are aggressive cancers associated with poor prognosis. BBC and GBM have stem cell‐like gene expression signatures, which are in part driven by forkhead box O (FOXO) transcription factors. To gain further insight into the impact of FOXO1 in BBC, we treated BT549 cells with AS1842856 and performed RNA sequencing. AS1842856 binds to unphosphorylated FOXO1 and inhibits its ability to directly bind to DNA. Gene Set Enrichment Analysis indicated that a set of WNT pathway target genes, including lymphoid enhancer‐binding factor 1 (LEF1) and transcription factor 7 (TCF7), were robustly induced after AS1842856 treatment. These same genes were also induced in GBM cell lines U87MG, LN18, LN229, A172, and DBTRG upon AS1842856 treatment. By contrast, follow‐up RNA interference (RNAi) targeting of FOXO1 led to reduced LEF1 and TCF7 gene expression in BT549 and U87MG cells. In agreement with RNAi experiments, CRISPR Cas9‐mediated FOXO1 disruption reduced the expression of canonical WNT genes LEF1 and TCF7 in U87MG cells. The loss of TCF7 gene expression in FOXO1 disruption mutants was restored by exogenous expression of the DNA‐binding‐deficient FOXO1‐H215R. Therefore, FOXO1 induces TCF7 in a DNA‐binding‐independent manner, similar to other published FOXO1‐activated genes such as TCF4 and hes family bHLH transcription factor 1. Our work demonstrates that FOXO1 promotes canonical WNT gene expression in examined BBC and GBM cells, similar to results found in Drosophila melanogaster, T‐cell development, and murine acute myeloid leukemia models.

Targeted refinement of regenerative materials requires mechanistic understanding of cell-material interactions. The nanoparticulate mineralized collagen glycosaminoglycan (MC-GAG) scaffold is shown to promote skull regeneration in vivo without additive exogenous growth factors or progenitor cells, suggesting potential for clinical translation. This work evaluates modulation of MC-GAG stiffness on canonical Wnt (cWnt) signaling. Primary human bone marrow-derived mesenchymal stem cells (hMSCs) are differentiated on two MC-GAG scaffolds (noncrosslinked, NX-MC, 0.3 kPa vs conventionally crosslinked, MC, 3.9 kPa). hMSCs increase expression of activated β-catenin, the major cWnt intracellular mediator, and the mechanosensitive YAP protein with near complete subcellular colocalization on stiffer MC scaffolds. Overall Wnt pathway inhibition reduces activated β-catenin and osteogenic differentiation, while elevating BMP4 and phosphorylated Smad1/5 (p-Smad1/5) expression on MC, but not NX-MC. Unlike Wnt pathway downregulation, isolated canonical Wnt inhibition with β-catenin knockdown increases osteogenic differentiation and mineralization specifically on the stiffer MC. β-catenin knockdown also increases p-Smad1/5, Runx2, and BMP4 expression only on the stiffer MC material. Thus, while stiffness-induced activation of the Wnt and mechanotransduction pathways promotes osteogenesis on MC-GAG, activated β-catenin is a limiting agent and may serve as a useful target or readout for optimal modulation of stiffness in skeletal regenerative materials.

Objective: To determine the diagnostic ability of Beta-catenin in poorly differentiated colorectal carcinomas. Study Design: Cross-sectional study Place and Duration of Study: Department of Histopathology PNS Shifa Hospital, Karachi Pakistan from Jun 2019 to Jun 2020 Methodology: After ethical approval from the Institutional Review Board, 60 patients of both gender and all age group, diagnosed with a case of colorectal carcinoma on biopsy and resection specimens analyzed on histopathology on H & E staining at PNS SHIFA were included in the study. The resection specimens of CRC included those obtained from hemicolectomy, abdominoperineal resection and biopsies. Beta-catenin was interpreted using immunohistochemistry as cytoplasmic and nuclear staining with varying intensity. The scoring method was 0 as negative, 1+ as weakly positive, 2+ as moderate positive, and 3+ as strongly positive. Results: Among the 60 patients with resection specimens included from hemicolectomy, abdominoperineal resection and colorectal biopsies, strong nuclear positive results were observed in 30(50 %) patients, moderate nuclear positivity in 12 (20 %), weak nuclear positivity in 6(10 %) and positive cytoplasmic staining in 12(20 %) patients. Conclusion: Beta-catenin can be used successfully as a diagnostic utility in poorly differentiated colorectal carcinoma patients.

BackgroundOverexpression of the transcription factor NR5A1 and constitutive activation of canonical Wnt signalling leading to nuclear translocation of beta-catenin are hallmarks of malignancy in adrenocortical carcinoma (ACC). Based on the analysis of genomic profiles in H295R ACC cells, Mohan et al. (Cancer Res. 2023; 83: 2123-2141) recently suggested that a major determinant driving proliferation and differentiation in malignant ACC is the interaction of NR5A1 and beta-catenin on chromatin to regulate gene expression.MethodsI reanalyzed the same set of data generated by Mohan et al. and other published data of knockdown-validated NR5A1 and beta-catenin target genes,ResultsBeta-catenin is mainly found in association to canonical T cell factor/lymphoid enhancer factor (TCF/LEF) motifs in genomic DNA. NR5A1 and beta-catenin regulate distinct target gene sets in ACC cells.ConclusionOverall, my analysis suggests a model where NR5A1 overexpression and beta-catenin activation principally act independently, rather than functionally interacting, to drive ACC malignancy.

ABSTRACTClassically, Axin1 is considered a regulator of Wnt/β-catenin signaling. However, Axin1’s roles in host-microbial interactions have been unknown. Our recent study has demonstrated that deletion of intestinal epithelial Axin1 in epithelial cells and Paneth cells protects the host against colitis by enhancing Akkermansia muciniphila. Loss of intestinal epithelial or Paneth cell Axin1 results in increased Wnt/β-catenin signaling, proliferation, and cell migration. This is associated with morphologically altered goblet and Paneth cells, including increased Muc2 and decreased lysozyme. Axin1 deletion specifically enriched Akkermansia muciniphila. Akkermansia muciniphila in Axin1 knockout mice is the driver of protection against DSS-induced inflammation. Here, we feature several significant conceptual changes, such as differences between Axin1 and Axin2, Axin1 in innate immunity and microbial homeostasis, and Axin1 reduction of Akkermansia muciniphila. We discuss an important trend in the field related to Paneth cells and tissue-specific Axin1 manipulation of microbiome in health and inflammation.

Background: Hematogenous tumor spread of malignant meningiomas occurs very rarely but is associated with very poor prognosis. Case presentation: We report an unusual case of a patient with a malignant meningioma who developed multiple metastases in bones, lungs and liver after initial complete resection of the primary tumor. After partial hepatic resection, specimens were histologically analyzed, and a complete loss of E-cadherin adhesion molecules was found. No oncogenic target mutations were found. The patient received a combination of conventional radiotherapy and peptide receptor radionuclide therapy (PRRT). Due to aggressive tumor behavior and rapid spread of metastases, the patient deceased after initiation of treatment. Conclusions: E-cadherin downregulation is associated with a higher probability of tumor invasion and distant metastasis formation in malignant meningioma. Up to now, the efficacy of systemic therapy, including PRRT, is very limited in malignant meningioma patients. [ABSTRACT FROM AUTHOR]

Pilomatricomas (PMs) are common benign adnexal tumors that show a predilection for the head and neck region and are characterized at the molecular level by activating mutations in the beta‐catenin (CTNNB1) gene. Giant PMs are a rare histopathological variant, according to the World Health Organization, which are defined by a size greater than 4 cm and are reported to show upregulation of yes‐associated protein compared to PMs of typical 1–3 cm size. We describe the case of a 67‐year‐old man with an 8 cm giant PM involving his temporal scalp, whose PM we characterized by 10X spatial gene expression analysis. This revealed five total transcriptomic clusters, including four distinct clusters within the giant PM, each with a unique transcriptional pattern of hair follicle‐related factors, keratin gene expression, and beta‐catenin pathway activity. [ABSTRACT FROM AUTHOR]

Basal‐like breast cancer (BBC) and glioblastoma multiforme (GBM) are aggressive cancers associated with poor prognosis. BBC and GBM have stem cell‐like gene expression signatures, which are in part driven by forkhead box O (FOXO) transcription factors. To gain further insight into the impact of FOXO1 in BBC, we treated BT549 cells with AS1842856 and performed RNA sequencing. AS1842856 binds to unphosphorylated FOXO1 and inhibits its ability to directly bind to DNA. Gene Set Enrichment Analysis indicated that a set of WNT pathway target genes, including lymphoid enhancer‐binding factor 1 (LEF1) and transcription factor 7 (TCF7), were robustly induced after AS1842856 treatment. These same genes were also induced in GBM cell lines U87MG, LN18, LN229, A172, and DBTRG upon AS1842856 treatment. By contrast, follow‐up RNA interference (RNAi) targeting of FOXO1 led to reduced LEF1 and TCF7 gene expression in BT549 and U87MG cells. In agreement with RNAi experiments, CRISPR Cas9‐mediated FOXO1 disruption reduced the expression of canonical WNT genes LEF1 and TCF7 in U87MG cells. The loss of TCF7 gene expression in FOXO1 disruption mutants was restored by exogenous expression of the DNA‐binding‐deficient FOXO1‐H215R. Therefore, FOXO1 induces TCF7 in a DNA‐binding‐independent manner, similar to other published FOXO1‐activated genes such as TCF4 and hes family bHLH transcription factor 1. Our work demonstrates that FOXO1 promotes canonical WNT gene expression in examined BBC and GBM cells, similar to results found in Drosophila melanogaster, T‐cell development, and murine acute myeloid leukemia models. [ABSTRACT FROM AUTHOR]

Background: The adrenal cortex provides adequate steroidogenic responses to environmental changes. However, in desert rodents, the adrenocortical activity varies according to several factors especially sex, age, and seasonal variations. Herein, we examined the sex differences in the adrenal cortex activity and explored the involvement of sex hormones in the regulation of this function in Libyan jird Meriones libycus. Materials and methods: Twenty-four adult male and female animals weighing 109–110 g were captured in the breeding season and equally assigned into control and gonadectomised groups. Animal euthanasia was performed 50 days after the gonadectomy. Adrenal gland was processed for structural and immunohistochemistry study of b-catenin, whereas plasma was used for cortisol assay. Results: The results showed that female adrenal gland weight was heavier than male and gonadectomy reduced this dimorphism. The adrenal cortex thickness was greater in the female than in the male, mainly due to significant development of the zona fasciculata. Females presented higher cell density in fasciculata and reticularis zones. The plasma cortisol was higher in females than in males. The immunolocalisation of beta-catenin showed that the expression was particularly glomerular in both sexes. However, in the female, the immunostaining was present in the zona reticularis while it was absent in the control male. Orchiectomy reduced zona glomerulosa cell density and induced hypertrophy of zona reticularis characterised by strong beta-catenin immunoreactivity. Conclusions: Results indicated that sex hormones had a major role in the regulation of the Saharan gerbil’s adrenal homeostasis by modulating beta-catenin signalling. Androgens seem to inhibit the Wnt-beta-catenin pathway and oestrogens are activators of the adrenal inner zones. [ABSTRACT FROM AUTHOR]

The Wnt signaling pathway is a highly conserved regulator of metazoan development and stem cell maintenance. Activation of Wnt signaling is an early step in diverse malignancies. Work over the past four decades has defined a "canonical" Wnt pathway that is initiated by Wnt proteins, secreted glycoproteins that bind to a surface receptor complex and activate intracellular signal transduction by inhibiting a catalytic complex composed of the classical tumor suppressor Adenomatous Polyposis Coli (APC), Axin, and Glycogen Synthase Kinase-3 (GSK-3). The best characterized effector of this complex is β-catenin, which is stabilized by inhibition of GSK-3, allowing β-catenin entrance to the nucleus and activation of Wnt target gene transcription, leading to multiple cancers when inappropriately activated. However, canonical Wnt signaling through the APC/Axin/GSK-3 complex impinges on other effectors, independently of β-catenin, including the mechanistic Target of Rapamycin (mTOR), regulators of protein stability, mitotic spindle orientation, and Hippo signaling. This review focuses on these alternative effectors of the canonical Wnt pathway and how they may contribute to cancers. [ABSTRACT FROM AUTHOR]

In melanoma, immune cell infiltration into the tumor is associated with better patient outcomes and response to immunotherapy. T‐cell non‐inflamed tumors (cold tumors) are associated with tumor cell‐intrinsic Wnt/β‐catenin activation, and are typically resistant to anti‐PD‐1 alone or in combination with anti‐CTLA‐4 therapy. Reversal of the 'cold tumor' phenotype and identifying new effective immunotherapies are challenges. We sought to investigate the role of a newer immunotherapy agent, B7‐H3, in this setting. RNA sequencing was used to identify co‐targeting strategies upon B7‐H3 inhibition in a well‐defined preclinical melanoma model driven by β‐catenin. We found that immune checkpoint molecule B7‐H3 confers a suppressive tumor microenvironment by modulating antiviral signals and innate immunity. B7‐H3 inhibition led to an inflamed microenvironment, up‐regulation of CD47/SIRPa signaling, and together with blockade of the macrophage checkpoint CD47 resulted in additive antitumor responses. We found that the antitumor effects of the B7‐H3/CD47 antibody combination were dependent on cytokine signaling pathways (CCR5/CCL5 and IL4). [ABSTRACT FROM AUTHOR]

Pancreatic solid pseudopapillary neoplasm (SPN) is a rare low grade malignant tumour. Distinguishing this entity from other pancreatic neoplasms is critical for therapeutic decision making and prognostication. It predominantly affects young female patients <40 years of age, with excellent clinical outcome following surgical removal. The gold standard diagnostic test is cytopathological or histopathological assessment of fine needle aspirate. There are two main difficulties with this. First, SPN can present with morphological and immunohistochemical appearances that can closely mimic other pancreatic tumours, in particular, pancreatic neuroendocrine tumour (NET). Second, the amount of diagnostic material from fine needle aspiration can be limited. Here, we present a cytopathological case with both challenges during the pre-operative investigation of SPN. The case exemplifies the importance of combining morphological features with a targeted panel of immunohistochemistry to arrive at the diagnosis. [ABSTRACT FROM AUTHOR]

Background: Kidney development is regulated by cellular interactions between the ureteric epithelium, mesenchyme, and stroma. Previous studies demonstrate essential roles for stromal β‐catenin in kidney development. However, how stromal β‐catenin regulates kidney development is not known. We hypothesize that stromal β‐catenin modulates pathways and genes that facilitate communications with neighboring cell populations to regulate kidney development. Results: We isolated purified stromal cells with wild type, deficient, and overexpressed β‐catenin by fluorescence‐activated cell sorting and conducted RNA Sequencing. A Gene Ontology network analysis demonstrated that stromal β‐catenin modulates key kidney developmental processes, including branching morphogenesis, nephrogenesis and vascular formation. Specific stromal β‐catenin candidate target genes that may mediate these effects included secreted, cell‐surface and transcriptional factors that regulate branching morphogenesis and nephrogenesis (Wnts, Bmp, Fgfr, Tcf/Lef) and secreted vascular guidance cues (Angpt1, VEGF, Sema3a). We validated established β‐catenin targets including Lef1 and novel candidate β‐catenin targets including Sema3e which have unknown roles in kidney development. Conclusions: These studies advance our understanding of gene and biological pathway dysregulation in the context of stromal β‐catenin misexpression during kidney development. Our findings suggest that during normal kidney development, stromal β‐catenin may regulate secreted and cell‐surface proteins to communicate with adjacent cell populations. Key Findings: β‐catenin in the renal stroma plays an essential role during kidney development. However, how stromal β‐catenin regulates kidney development is unknown. We identified differentially expressed genes and biological pathways in the renal stroma in mouse models with stabilized or deficient stromal β‐catenin. Our data suggest that stromal β‐catenin may regulate branching morphogenesis, nephrogenesis and vascular development through the regulation of secreted and cell‐surface proteins to facilitate cell‐to‐cell communication during development. Among the differentially expressed genes, we identified and validated candidate target genes that have known roles in kidney development, and other novel targets with unexplored roles. [ABSTRACT FROM AUTHOR]

The adherens junctions (AJs) maintain the epithelial cell layers' structural integrity and barrier function. AJs also play a vital role in various biological and pathological processes. AJs perform these functions through the cadherin-catenin adhesion complex. This study investigated the presence, cell-specific localization, and temporal distribution of AJ components such as classical type I cadherins and beta-catenin in the cow cervix and vagina during the estrous cycle. Immunohistochemistry and Western blot analysis results demonstrated that beta-catenin and epithelial (E)-, neural (N)-, and placental (P)-cadherins are expressed in the cow cervix and vagina during the estrous cycle. These adhesion molecules were localized in the membrane and cytoplasm of the ciliated and non-ciliated cervical cells and the stratified vaginal epithelial cells. Positive immunostaining for P-, N-cadherin, and beta-catenin was also observed in the vascular endothelial cells of the cervical and vaginal stroma. Quantitative immunohistochemistry examinations revealed that in the cervical and vaginal epithelia, P-cadherin's optical density values (ODv) were the highest; in contrast, the N-cadherin ODv were the lowest. The ODv of P-cadherin and beta-catenin in the cervical epithelium and E-cadherin in the vagina were significantly higher in the luteal phase versus the follicular phase of the estrous cycle. Furthermore, the ODv of P-cadherin, N-cadherin, and beta-catenin in the cervix's central and peripheral epithelial regions were different during the estrous cycle. These findings indicate that classical cadherins and beta-catenin in the cervix and vagina exhibit cell- and tissue-specific expression patterns under the influence of estrogen and progesterone hormones during the estrous cycle. [ABSTRACT FROM AUTHOR]

Desmoid tumour is a rare, benign soft tissue tumour. It is noted to be clinically aggressive but pathologically benign, showing a high local recurrence rate; however, it cannot metastasize. Although it is benign, it can impact functionality. The majority of these cases remain sporadic, and the aetiology is unknown. The retroperitoneum is a rare site, and there are few reports of desmoidtype fibromatosis occurring in this region. It is difficult to distinguish from other tumours and to identify the tumour’s origin. Here, the authors present a case of a 36-year-old female who presented with abdominal pain and a mass in her abdomen. Imaging studies revealed a large retroperitoneal mass extending from the L5 vertebral body up to the inferior endplate of the L1 vertebral body. Exploratory laparotomy was performed, and the retroperitoneal mass was resected. Grossly, it was a capsulated dark brown to grey-coloured mass measuring 15×12×10 cm. The mass was sent for histopathology {Haematoxylin and Eosin (H&E)} study and reported as a retroperitoneal desmoid tumour, which was further confirmed on Immunohistochemistry (IHC) by showing immunopositivity for Desmin, Muscle-Specific Actin (MSA), and Beta-catenin. The patient is doing well with no further complaints to date. No disease relapse has occurred. Follow-up has been done every three months till the writing of the present report.

Background & aimsGain of function (GOF) mutations in the CTNNB1 gene are one of the most frequent genetic events in hepatocellular carcinoma (HCC). T-box transcription factor 3 (TBX3) is a liver-specific target of the Wnt/β-catenin pathway and thought to be an oncogene mediating activated β-catenin-driven HCC formation.MethodsWe evaluated the expression pattern of TBX3 in human HCC specimens. Tbx3 was conditionally knocked out in murine HCC models by hydrodynamic tail vein injection of Cre together with c-Met and ΔN90-β-catenin (c-Met/β-catenin) in Tbx3flox/flox mice. TBX3 was overexpressed in human HCC cell lines to investigate the functions of TBX3 in vitro.ResultsA bimodal expression pattern of TBX3 in human HCC samples was detected: high expression of TBX3 in GOF CTNNB1 HCC and downregulation of TBX3 in non-CTNNB1 mutant tumors. High expression of TBX3 was associated with increased differentiation and decreased expression signatures of tumor growth. Using Tbx3flox/flox mice, we found that ablation of Tbx3 significantly accelerates c-Met/β-catenin-driven HCC formation. Moreover, Tbx3(-) HCC demonstrated increased YAP/TAZ activity. The accelerated tumor growth induced by loss of TBX3 in c-Met/β-catenin mouse HCC was successfully prevented by overexpression of LATS2, which inhibited YAP/TAZ activity. In human HCC cell lines, overexpression of TBX3 inhibited HCC cell growth as well as YAP/TAZ activation. A negative correlation between TBX3 and YAP/TAZ target genes was observed in human HCC samples. Mechanistically, phospholipase D1 (PLD1), a known positive regulator of YAP/TAZ, was identified as a novel transcriptional target repressed by TBX3.ConclusionOur study suggests that TBX3 is induced by GOF CTNNB1 mutants and suppresses HCC growth by inactivating PLD1, thus leading to the inhibition of YAP/TAZ oncogenes.Lay summaryTBX3 is a liver-specific target of the Wnt/β-catenin pathway and thought to be an oncogene in promoting liver cancer development. Herein, we demonstrate that TBX3 is in fact a tumor suppressor gene that restricts liver tumor growth. Strategies which increase TBX3 expression and/or activities may be effective for HCC treatment.

Liver fibrosis occurs in response to chronic liver injury and is characterized by the production of excess extracellular matrix (ECM) proteins, largely by activated hepatic stellate cells (HSCs). Numerous studies have implicated microribonucleic acids (miRNAs) in liver fibrosis, but the mechanisms remain unclear. Herein, HSC activation by miR-33a-5p during hepatic fibrosis was investigated. The miR-33a-5p was increased in the fibrotic mice and activated HSCs. AntagomiR-33a-5p inhibited HSC activation, proliferation, and migration in vitro, while simultaneously inducing apoptosis. The luciferase reporter assays indicated that the miR-33a-5p bound to the three prime untranslated region (3'UTR) of Dickkopf-1 (DKK1). Further investigation revealed that the miR-33a-5p targeted DKK1-modulated wingless-related integration site (Wnt)/b-catenin signaling directly to control hepatic fibrosis. Notably, the mice treated with antgomiR-33a-5p exhibited increased expression of DKK1 and reduced expression of fibrosis markers, along with reduced fibrosis. The RNA was isolated from activated and quiescent LX-2 cells and subsequently sequenced. Transcriptomic and bioinformatic analyses indicated strong downregulation of DKK1 during LX-2 cell activation. This paper presents the first demonstration of the miR-33a-5p-mediated modulation of liver fibrosis, with miR-33a-5p found to interact with DKK1, leading to regulation of Wnt/b-catenin signaling. The transcriptomic changes occurring during HSC activation were also defined. Overall, the findings suggest that both miR-33a-5p and DKK1 may be useful targets for treating liver fibrosis. [ABSTRACT FROM AUTHOR]

Expanding antiviral treatment options against SARS-CoV-2 remains crucial as the virus evolves under selection pressure which already led to the emergence of several drug resistant strains. Broad spectrum host-directed antivirals (HDA) are promising therapeutic options, however the robust identification of relevant host factors by CRISPR/Cas9 or RNA interference screens remains challenging due to low consistency in the resulting hits. To address this issue, we employed machine learning, based on experimental data from several knockout screens and a drug screen. We trained classifiers using genes essential for virus life cycle obtained from the knockout screens. The machines based their predictions on features describing cellular localization, protein domains, annotated gene sets from Gene Ontology, gene and protein sequences, and experimental data from proteomics, phospho-proteomics, protein interaction and transcriptomic profiles of SARSCoV-2 infected cells. The models reached a remarkable performance suggesting patterns of intrinsic data consistency. The predicted HDF were enriched in sets of genes particularly encoding development, morphogenesis, and neural processes. Focusing on development and morphogenesis-associated gene sets, we found β-catenin to be central and selected PRI-724, a canonical β-catenin/CBP disruptor, as a potential HDA. PRI-724 limited infection with SARS-CoV-2 variants, SARS-CoV-1, MERS-CoV and IAV in different cell line models. We detected a concentration-dependent reduction in cytopathic effects, viral RNA replication, and infectious virus production in SARS-CoV-2 and SARS-CoV-1-infected cells. Independent of virus infection, PRI-724 treatment caused cell cycle deregulation which substantiates its potential as a broad spectrum antiviral. Our proposed machine learning concept supports focusing and accelerating the discovery of host dependency factors and identification of potential host-directed antivirals. [ABSTRACT FROM AUTHOR]

Cross talk between immune cells and the intestinal crypt is critical in maintaining intestinal homeostasis. Recent studies highlight the direct impact of vitamin D receptor (VDR) signaling on intestinal and microbial homeostasis. However, the tissue‐specific role of immune VDR signaling is not fully understood. Here, we generated a myeloid‐specific VDR knockout (VDRΔLyz) mouse model and used a macrophage/enteroids coculture system to examine tissue‐specific VDR signaling in intestinal homeostasis. VDRΔLyz mice exhibited small intestine elongation and impaired Paneth cell in maturation and localization. Coculture of enteroids with VDR−/− macrophages increased the delocalization of Paneth cells. VDRΔLyz mice exhibited significant changes in the microbiota taxonomic and functional files, and susceptibility to Salmonella infection. Interestingly, loss of myeloid VDR impaired Wnt secretion in macrophages, thus inhibiting crypt β‐catenin signaling and disrupting Paneth cell differentiation in the epithelium. Taken together, our data have demonstrated that myeloid cells regulate crypt differentiation and the microbiota in a VDR‐dependent mechanism. Dysregulation of myeloid VDR led to high risks of colitis‐associated diseases. Our study provided insight into the mechanism of immune/Paneth cell cross talk in regulating intestinal homeostasis. [ABSTRACT FROM AUTHOR]