E-cadherin Expression Research Articles

MSC-exosomes pretreated by Danshensu extracts pretreating to target the hsa-miR-27a-5p and STAT3-SHANK2 to enhanced antifibrotic therapy

BackgroundPeritoneal fibrosis (PF) is a serious complication commonly associated with prolonged peritoneal dialysis. Mesenchymal stem cells (MSCs) and their exosomes (Exo) have shown significant therapeutic promise in treating fibrotic conditions. Danshensu (DSS), a bioactive compound from the traditional Chinese herb Danshen reverses fibrosis. This study aims to investigate a novel strategy to enhance the therapeutic efficacy against PF by DSS preconditioning MSCs-derived exosomes (DSS-Exo).MethodsThe in vitro studies included the effects of DSS duration on MSCs, and the characterization of DSS-Exo and Exo, followed by the assessment of RNA and protein expression levels of peritoneal fibrosis markers and inflammatory cytokines levels after treating human peritoneal mesothelial (HMrSV5) cells. In vivo experiments were conducted on a PF mouse model to observe cell morphology, collagen deposition, fibrosis localization, and to evaluate peritoneal functions such as filtration rate, urea nitrogen clearance, peritoneal thickness, and protein leakage. Mechanistic insights were gained through the analysis of the STAT3/HIF-1α/VEGF signaling pathway, tissue dual-fluorescence localization,chromatin immunoprecipitation sequencing (ChIP-seq), and dual-luciferase reporter (DLR) assays. Additionally, the differential expression of miRNAs between DSS-Exo and Exo was explored and validation of key miRNA.ResultsDSS-Exo significantly upregulated E-cadherin, downregulated VEGFA, α-SMA, CTGF and Fibronectin expression in HMrSV5 cells compared to untreated Exo. In vivo studies revealed that DSS-Exo enhanced the ability of Exo to improve peritoneal function,such as the peritoneal filtration rate and urea nitrogen, glucose clearance, while reducing peritoneal thickness and protein leakage, and cell morphology, reduce collagen deposition, and decrease the degree of fibrosis. Mechanistically, these exosomes inhibited the STAT3/HIF-1α/VEGF signaling pathway within peritoneal mesothelial tissues. Furthermore, ChIP-seq and DLR demonstrated that DSS-Exo affected STAT3 directly binds to SHANK2 promoter regions, forming hydrogen bonds between 5 key amino acids such as GLN-344, HIS-332 and 6 key bases such as DG-258, DG-261. miRNA profiling identified DSS-Exo increased hsa-miR-27a-5p_R-1 to regulated STAT3-SHANK2 and modulating the EMT.ConclusionThis study highlighted the innovative use of Danshensu in enhancing MSC-derived exosome therapy for PF. The identification of the hsa-miR-27a-5p_R-1-STAT3-SHANK2 axis may reveal new molecular mechanisms underlying fibrosis, further research is needed to fully elucidate its impact on PF. The integration of Danshensu from traditional Chinese medicine into modern MSC exosome therapy represents a promising frontier in the development of novel treatments for fibrotic diseases.

Plasticity of Expression of Stem Cell and EMT Markers in Breast Cancer Cells in 2D and 3D Culture Depend on the Spatial Parameters of Cell Growth; Mathematical Modeling of Mechanical Stress in Cell Culture in Relation to ECM Stiffness

The majority of the current cancer research is based on two-dimensional cell cultures and animal models. These methods have limitations, including different expressions of key factors involved in carcinogenesis and metastasis, depending on culture conditions. Addressing these differences is crucial in obtaining physiologically relevant models. In this manuscript we analyzed the plasticity of the expression of stem cell and epithelial/mesenchymal markers in breast cancer cells, depending on culture conditions. Significant differences in marker expression were observed in different growth models not only between 2D and 3D conditions but also between two different 3D models. Differences observed in the levels of adherent junction protein E-cadherin in two different 3D models suggest that spatial parameters of cell growth and physical stress in the culture may affect the expression of junction proteins. To provide an explanation of this phenomenon on the grounds of mechanobiology, these parameters were analyzed using a mathematical model of the 3D bioprinted cell culture. The finite element mechanical model generated in this study includes an extracellular matrix and a group of regularly placed cells. The single-cell model comprises an idealized cytoskeleton, cortex, cytoplasm, and nucleus. The analysis of the model revealed that the stress generated by external pressure is transferred between the cells, generating specific stress fields, depending on growth conditions. We have analyzed and compared stress fields in two different growth conditions, each corresponding to a different elasticity of extracellular matrix. We have demonstrated that soft matrix conditions produce more stress than a stiff matrix in the single cell as well as in cellular spheroids. The observed differences can explain the plasticity of E-cadherin expression in response to mechanical stress. These results should contribute to a better understanding of the differences between various growth models.

Yiqi Juanshen decoction alleviates renal interstitial fibrosis by targeting the LOXL2/PI3K/AKT pathway to suppress EMT and inflammation

Chronic kidney disease (CKD) is a major health concern, with renal interstitial fibrosis (RIF) as a key feature. Effective management of RIF is crucial for treating CKD. Yiqi Juanshen decoction (YQJSD), as traditional Chinese medicine, has shown promising results in CKD treatment. This study evaluates YQJSD’s effectiveness in ameliorating RIF and explores the underlying molecular mechanisms using the unilateral ureteral obstruction (UUO) model. YQJSD has been shown to effectively reduce serum creatinine and blood urea nitrogen levels, decrease extracellular matrix deposition, and down-regulate the expression of α-SMA, COL4α1, Fibronectin (FN). Mechanistically, YQJSD exerts its effects by modulating multiple pathways: it inhibits the NF-κB signaling pathway, inhibiting the expression of pro-inflammatory cytokines like NF-κB1, IL-1β, TNF-α, and CCR1. Simultaneously, YQJSD suppresses the epithelial-mesenchymal transition (EMT) by downregulating the expression of Snail1, Vimentin, Twist1, and FSP1, while increasing E-cadherin expression. Moreover, YQJSD can regulate the PI3K/AKT signaling pathway by decreasing the expression of LOXL2 and PIK3R1, along with p-AKT1/2/3. This modulation of the LOXL2/PI3K/AKT pathway contributes to the inhibition of both EMT and inflammation, highlighting a critical role in the therapeutic intervention against RIF. These findings suggest that YQJSD may serve as a promising therapeutic management of RIF in CKD patients.

Role of 11β-hydroxysteroid dehydrogenase type 1 inhibition in the antiobesity effect of J2H-1702 on adipocytes and a high-fat diet-induced NASH model.

Obesity due to excessive body fat accumulation remains a global problem. Patients with obesity have high cortisol levels, and its dysregulation is caused by increased 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) levels. The effects and mechanism of J2H-1702, an 11β-HSD1 inhibitor, on nonalcoholic steatohepatitis (NASH) were explored. This study compared the antiadipogenic effects of J2H-1702, elafibranor (PPARα/δ agonist), and BVT14225 (selective 11β-HSD1 inhibitor) using mouse 3T3-L1 pre-adipocytes. J2H-1702, elafibranor, and BVT14225 inhibited adipocyte differentiation and intracellular lipid accumulation in 3T3-L1 cells by downregulating phospho-extracellular signal-regulated kinase, extracellular signal-regulated kinase, phospho-c-Jun-N-terminal Kinase, c-Jun-N-terminal Kinase, phospho-P38 (P-P38), P38, CCAAT/enhancer-binding proteins alpha and β, peroxisome proliferator-activated receptor γ, and glucocorticoid receptor. Additionally, J2H-1702, elafibranor, and BVT14225 treatments effectively inhibited 11β-HSD1 activity, as revealed by cortisol concentrations, and inhibited cortisone-induced adipocyte differentiation and intracellular lipid accumulation in 3T3-L1 cells. These effects were associated with 11β-HSD1 protein inhibition. Furthermore, J2H-1702 and BVT14225 increased the expression of Akt and phosphoinositide 3-kinase involved in insulin resistance in 3T3L-1 adipocytes. In the LX-2 human hepatic stellate cell line, the relative expression of N-cadherin, 11β-HSD1, collagen1α (COLA1), α-actin of smooth muscle (α-SMA) genes in LX-2 activated with TGF-β increased significantly, and after treatment with J2H-1702, it was significantly reduced. The expression of E-cadherin is decreased in TGF-β-treated LX-2 cells and increased after treatment with J2H-1702. We tested the potential of J2H-1702 as a therapeutic agent for NASH using a high-fat diet-induced NASH model, with obeticholic acid, an FXR agonist, and elafibranor as reference drugs. All drugs significantly decreased the elevated triglyceride levels in the livers of high-fat, high-carbohydrate (HFHC-fed mice. The results may add to the benefits of targeting 11β-HSD1 inhibitors with antiadipogenic activity in developing a therapeutic agent for obesity treatment.

High mobility group protein N2 inhibits the progression of hepatocellular carcinoma and the related molecular mechanisms.

High mobility group protein N2 (HMGN2) related pathways are involved in chromatin regulation/acetylation. It has been reported to be involved in several types of cancers. A recent sequencing study suggested that HMGN2 might be involved in the progression of hepatocellular carcinoma (HCC). This study aimed to explore the role of HMGN2 in HCC, which has been proven to be involved in the development of HCC. In this study, we collected clinical samples and cultured normal hepatocytes and hepatocellular carcinoma cell lines to detect HMGN2 expression levels using reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) and western blot assay. Subsequently, to determine the role of HMGN2 in HCC, HMGN2 was overexpressed in HCC cell lines. MTT (3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di- phenytetrazoliumromide) assay was used to detect the cell proliferative capacity, and proliferation-related proteins were detected by RT-qPCR and western blot assay. To observe the effect of HMGN2 on cell migration and invasion capacity, Transwell assay was performed. Then, cell apoptosis was detected by flow cytometry, and caspase3 and cleaved-caspase3 were detected using western blot assay. Finally, EMT (epithelial to mesenchymal transition)-related proteins, and matrix metalloproteinase-2 (MMP-2) and MMP-9 expression were detected by RT-qPCR and western blot assay. HMGN2 expression was decreased in HCC tissues as well as in HCC cell lines. After overexpression of HMGN2, MTT results suggested that cell proliferation was decreased, and flow cytometry results showed that the apoptosis level was increased and ki-67 and proliferating cell nuclear antigen (PCNA) expression levels were decreased. On the contrary, cleaved-caspase 3 expression level was increased. HCC cells overexpressing HMGN2 showed a drastic reduction in the number of migrating and invading cells, and the expression levels of MMP-2 and MMP-9 were significantly decreased. Finally, E-cadherin expression was elevated in HCC cells transfected with the HMGN2-plasmid, while N-cadherin showed the opposite result. HMGN2 expression was significantly decreased in patients with HCC. HMGN2 inhibits the malignant behavior of HCC cells and is a potential therapeutic target for HCC.

Molecular Dynamics of Breast Cancer Subtypes: The Role of FAM83H-AS1 Long Non-coding RNA in Breast Cancer Metastasis.

Breast cancer is the leading cause of cancer-related deaths in women. Long non-coding RNAs (lncRNAs) play an important role in gene regulation and are emerging as major players in cancer biology, This study investigates the expression of FAM83H-AS1 in breast cancer and its association with tumor grade, hormone receptors, pathological diagnosis, and molecular markers related to epithelial-mesenchymal transition (EMT). The expression of the long non-coding RNA FAM83H-AS1 in 80 breast cancer patients was assessed using quantitative real-time PCR (qRT-PCR). Clinical significance was evaluated through histopathological and immunohistochemical analyses. The associations of FAM83H-AS1 expression with tumor grade, hormone receptor status, and epithelial-mesenchymal transition (EMT) markers were analyzed. A positive correlation was observed between tumor grade and the expression of FAM83H-AS1, N-cadherin, E-cadherin, and vimentin, whereas FGF-18, TGF-β, and β-catenin were downregulated. Estrogen receptor positivity was associated with CLDN1 and Snail-1 expression, while HER2 positivity was linked to vimentin expression. Snail-1 expression correlated positively with Ki-67 levels. All genes except MMP2 were significantly associated with lymph node metastasis. Comparative analysis revealed significant differences in FGF-18, TGF-β, N-cadherin, β-catenin, and MMP2 expression among luminal A, luminal B, and triple-negative breast cancer (TNBC) subtypes. FAM83H-AS1 was upregulated in TNBC compared to luminal A and inflammatory breast cancer (IBC), although the difference was not statistically significant. TNBC Exhibited upregulation of TGF-β, N-cadherin, and β-catenin, suggesting their role in the aggressive nature of this subtype. In contrast, MMP2 was downregulated in TNBC compared to IBC, potentially indicating a suppressive role in tumor invasion in TNBC. Vimentin was upregulated in IBC compared to luminal A, indicating its involvement in IBC's aggressive behavior. MMP2 and MMP9 were significantly upregulated in IBC compared to luminal A. FAM83H-AS1 shows potential as a prognostic biomarker and therapeutic target, especially in TNBC and IBC, with implications for personalized breast cancer treatment strategies. Its expression correlates with tumor grade, hormone receptor status, and EMT markers, suggesting a role in cancer progression and metastasis.

KDM1A Acts as an Oncogene and Facilitates the Epithelial Mesenchymal Transition Process in Gastric Cancer.

This study is performed to research the biological role of KDM1A in the epithelial mesenchymal transition (EMT) of gastric cancer and investigate the mechanism involved. The KDM1A, Vimentin and E-cadherin levels were studied, as well as the correlation among them in gastric cancer samples. Gastric cancer cells were transfected with KDM1A overexpression and knockdown, and the cellular infiltration, motility, morphology and F-actin expression were subsequently identified. For the protein level assessment of EMT, the western blot analysis combined with immunofluorescence was employed. The effect of KDM1A on TGF-β/Notch signaling was also detected. KDM1A was overexpressed in gastric cancer tumor tissues. In the clinical gastric carcinoma samples, the level of KDM1A was linked negatively to the expression of E-cadherin, while positively to the expression of Vimentin. Among the gastric carcinoma population, the expression of KDM1A was linked to the lymph node metastasis, TNM stage and tumor differentiation. The KDM1A downregulation prohibited the cellular motility, infiltration and F-actin expression, and suppressed EMT process. KDM1A overexpression exhibited promoting effect on EMT in gastric cancer cells. KDM1A regulated TGF-β/Notch signaling to affect EMT in gastric cancer cells. KDM1A acts as an oncogene and facilitates the epithelial mesenchymal transition process by regulating TGF-β/Notch signal pathway in gastric cancer cells.

Upregulation of the MAP2K4 gene triggers endothelial-mesenchymal transition in COVID-19.

SARS-CoV-2 infection is marked by an excessive inflammatory response, leading to elevated production of pro-inflammatory cytokines through activation of intracellular pathways like mitogen-activated protein kinase (MAPK). Viruses can use the MAPK signaling pathway to their advantage, but the relationship of this pathway to the severe SARS-CoV-2 period has not been fully elucidated. MAP2K4 is involved in the MAPK signaling pathway and affects cellular processes such as cell-cell junction, cell proliferation, differentiation and apoptosis. In this study, we sought to determine the associated biomarkers that are involved in the MAP2K4 pathway and elucidate its possible roles in terms of some clinical features associated with COVID-19. We evaluated the expressions of MAP2K4, SNAI1, SLUG, ZEB1 and E-Cadherin. For this purpose, we prospectively recruited 66 individuals, 39 of whom were women and had a mean age of 65 years. The results revealed that MAP2K4 upregulation increased SNAI1 gene expression level whereas E- Cadherin level was decreased in SARS-CoV-2 positive participants. In addition, negative correlations were determined with PLT, Lymphocyte and CKMB and E- Cadherin levels in positive participants. We also observed a negative correlation between the MAP2K4 and AST, and a positive correlation between SLUG and BUN, ZEB1 and CK. We conclude that SARS-CoV-2 infection triggers fibrosis by increasing MAP2K4 regulation. Additionally, this is the first study to demonstrate the possible contribution of MAP2K4 in influencing COVID-19 clinical features, which may be relevant for identifying COVID-19 positive participants with severe complications.

Real-world of Limosilactobacillus reuteri in mitigation of acute experimental colitis

Probiotics have been proposed as a potential strategy for managing ulcerative colitis (UC). However, the underlying mechanisms mediating microbiota-host crosstalk remain largely elusive. Here, we report that Limosilactobacillus reuteri (L. reuteri), as a probiotic, secretes cytoplasmic membrane vesicles (CMVs) that communicate with host cells, alter host physiology, and alleviate dextran sulfate sodium (DSS)-induced colitis. First, L. reuteri-CMVs selectively promoted the proliferation of the beneficial bacterium Akkermansia muciniphila (AKK) by upregulating the expression of glycosidases (beta-N-acetylhexosaminidase and alpha-N-acetylglucosaminidase) involved in glycan degradation and metabolic pathways and restored the disrupted gut microbiota balance. Second, L. reuteri-CMVs were taken up by intestinal epithelial cells (IECs), elevated the expression of ZO-1, E-cadherin (Cdh1), and Occludin (Ocln), decreased intestinal permeability, and exerted protective effects on epithelial tight junction functionality. RNA sequencing analysis demonstrated that L. reuteri-CMVs repaired intestinal barrier by activating the HIF-1 signaling pathway and upregulating HMOX1 expression. Third, L. reuteri-CMVs increased the population of double positive (DP) CD4+CD8+ T cells in the intestinal epithelial layer, suppressing gut inflammation and maintaining gut mucosal homeostasis. Finally, L. reuteri-CMVs exhibited satisfactory stability and safety in the gastrointestinal tract and specifically targeted the desired sites in colitis mice. Collectively, these findings shed light on how L. reuteri interact with the host in colitis, and provide new insights into potential strategies for alleviating colitis.Graphical

Monotropein represses the proliferation and angiogenesis via the AKT/NF-κB pathway in lung cancer.

Monotropein (Mon) is an iridoid glycosides extracted from Morinda officinalis F.C. How. (Rubiaceae) that shows anticancer effects on colorectal cancer. This study aimed to investigate the therapeutic effect of Mon on lung cancer. The viability, apoptosis, invasion, and tube formation of A549 and H1299 were determined by CCK8 assay, flow cytometry, transwell assay, and tube formation assay. NF-κB expression in the nucleus was detected by immunofluorescent staining. In vivo assay, BALB/c nude mice were divided into a sham group and a 2mg/kg Mon group. For the Mon group, mice were orally administered with 2mg/kg Mon. The duration of the animal study was 35days, and the tumor formation and angiogenesis were investigated. Compared with the control (Mon 0μM) group, Mon dramatically repressed the viability, invasion, tube formation, proliferation marker Ki67, N-cadherin, and VEGFA expressions in A549 cells (Mon of 25, 50, and 100µM) and H1299 cells (Mon of 50 and 100µM). Mon dramatically promoted cell apoptosis, cleaved caspase 3, and E-cadherin expressions. Besides, Mon remarkably downregulated p-AKT and p-NF-κB protein expressions. Moreover, AKT agonist SC79 reversed the anticancer effects of 100µM Mon on A549 cells. Furthermore, Mon markedly suppressed tumor growth, decreased N-cadherin, VEGFA, p-AKT, and p-NF-κB expressions, increased apoptosis and E-cadherin expression, and SC79 reversed the inhibition effects of Mon in vivo. Thus, Mon is a potential antitumor natural drug, and more signaling pathways involved in Mon-modulated lung cancer progression are worth testing for further investigation.

Integrating E-cadherin expression levels with TNM staging for enhanced prognostic prediction in colorectal cancer patients

PurposeThis study aimed to investigate the clinical significance of E-cadherin expression levels in colorectal cancer tissues and explore the relationship between E-cadherin expression and tumor node metastasis (TNM) stage. The goal was to establish a more accurate prognostic prediction for colorectal cancer patients by analyzing E-cadherin expression levels alongside TNM staging.MethodsThe study examined colorectal cancer patients by dividing them into groups based on E-cadherin expression levels. It then assessed their 5-year event-free survival (EFS) and disease-specific survival (DSS) hazard ratios (HRs). Additionally, the prognosis of patients was analyzed by combining E-cadherin expression levels with TNM staging, particularly focusing on patients in stages III and IV.ResultsThe E-cadherinLow group had significantly worse outcomes, with HRs of 2.30 for EFS and 2.76 for DSS compared to the E-cadherinHigh group. When combined with TNM stage III/IV, patients with E-cadherinLow expression showed a poor prognosis, with HRs of 1.93 for EFS and 2.35 for DSS compared to those with E-cadherinHigh expression at the same TNM stage.ConclusionsLow levels of E-cadherin expression are associated with a poor prognosis and decreased survival in colorectal cancer patients. Combining E-cadherin expression levels with TNM staging provides a more precise prediction of patient prognosis and survival, potentially guiding personalized treatment strategies.

Loss of PRICKLE1 leads to abnormal endometrial epithelial architecture, decreased embryo implantation, and reduced fertility in mice

Abstract Successful embryo implantation requires coordinated changes in the uterine luminal epithelium, including structural adaptations, apical-basal polarity shifts, intrauterine fluid resorption, and cellular communication. Planar cell polarity (PCP) proteins, essential for cell organization, are understudied in the context of uterine physiology and implantation. PRICKLE proteins, components of PCP, are suggested to play critical roles in epithelial polarization and tissue morphogenesis. However, their function in the polarized unicellular layer of endometrial epithelium, which supports embryo implantation, is unknown. We developed an endometrial epithelial-specific knockout (cKO) of mouse Prickle1 using Lactoferrin-iCre to investigate its’s role in uterine physiology. Prickle1 ablation in the endometrial epithelium of mice resulted in decreased embryo implantation by gestational day 4.5 leading to lower fertility. Three-dimensional imaging of the uterus revealed abnormal luminal folding, impaired luminal closure, and altered glandular length in mutant uteri. Additionally, we observed decreased aquaporin-2 expression, disrupted cellular architecture, and altered E-Cadherin expression and localization in the mutant uterine epithelium. Evidence of epithelial-mesenchymal transition (EMT) was found within luminal epithelial cells, further linking PRICKLE1 loss to uterine pathologies. Furthermore, altered polarity of cell division leading to incomplete cytokinesis and increase in binuclear or multinucleated cells suggests a crucial role for PRICKLE1 in the maintenance of epithelial architecture. Our findings highlight PRICKLE1's critical role in the PCP pathway within the uterus, revealing its importance in the molecular and cellular responses essential for successful pregnancy and fertility.

Prognostic Implications of Decorin, E-Cadherin and EGFR Expression in Inflammatory and Non-Inflammatory Canine Mammary Carcinomas.

Inflammatory mammary carcinoma (IMC) is the most aggressive variant of invasive mammary tumours in dogs and in women. Decorin is an extracellular matrix molecule whose expression can be reduced or absent in various human cancers, which is associated with a poor prognosis. E-cadherin is a cell adhesion protein whose expression is reduced in several neoplasms. However, it is overexpressed in inflammatory breast cancers of women. EGFR is also associated with cancer development and is commonly overexpressed in aggressive neoplasms. This study aimed to characterise the expressions of Decorin, E-cadherin, and EGFR in canine inflammatory and non-inflammatory mammary carcinomas (IMC and non-IMC) and to evaluate their expression levels as prognostic indicators for survival and occurrence of metastases. Thirty-three IMC and 43 non-IMC cases were analysed retrospectively and submitted to immunohistochemical analysis. The reactions were quantified in five high-power field images from areas of the highest intensity and frequency of immunostaining (hot spots). We found significantly lower expression of Decorin and higher of E-cadherin and EGFR in canine IMCs. Patients with tumours that exhibited Decorin expression in less than 26.35% of epithelial cells had shorter survival (p = 0.0410) and a higher occurrence of distant metastases (p = 0.0115). E-cadherin is overexpressed in canine IMCs (p < 0.0001), similar to what occurs in women, reinforcing that dogs can be used as a study model for human IMC. EGFR overexpression in canine IMCs (p = 0.0322) provides evidence for potential targeted therapy with tyrosine kinase inhibitors.

P0213 Salivary Veillonella parvula from Crohn’s Disease Patients Exacerbates Intestinal Inflammation

Abstract Background The gut microbiota plays a significant role in the development of inflammatory bowel disease (IBD). There is a close interaction between the oral and gut microbiota. However, the role of the "oral-gut" microbial axis in the development of IBD is not yet clear. Methods 1. Analyzed oral-gut microbiota patterns in 97 IBD patients using 16S rDNA sequencing and bioinformatics methods. 2. A mouse colitis model with DSS was used. It was gavaged with oral microbiota from CD patients to assess the impacts on colitis and gut microbiota. 3. Isolated specific differential bacteria, Veillonella, from CD patient saliva and evaluated their effects on DSS-induced colitis in mice. Results 1. IBD patients had a reduced core gut microbiota, lower diversity (P &amp;lt; 0.05), and increased abundance of potential pathogens such as Escherichia-Shigella. 2. CD patients showed more correlations between saliva and fecal microbiota, with the most significant correlation between Veillonella in saliva and Escherichia-Shigella in fecal (P=0.00018) . 3. The mice which were gavaged with saliva from CD patients showed significant weight loss (P &amp;lt; 0.0001), reduced colon length (P=0.03), increased histopathological scores (P=0.01), decreased expression of colonic mucosal barrier proteins ZO-1 and E-cadherin, and exacerbated gut microbiota dysbiosis . 4. The mice which were gavaged with Veillonella parvula from CD patients showed significant weight loss (P &amp;lt; 0.0001), reduced colon length (P=0.02), increased histopathological scores (P=0.02), decreased expression of colonic mucosal barrier proteins ZO-1 and E-cadherin, and exacerbated gut microbiota dysbiosis. Conclusion The interplay between the oral and gut microbiota potentially influences the progression of IBD, with oral bacteria such as Veillonella possibly intensifying intestinal inflammation in IBD patients by disrupting the gut microbiota balance. Targeted modulation of the gut microbiota and interference with oral-gut microbial interactions may offer a promising therapeutic strategy for managing IBD.

Clinical correlation of p16 expression with lymphatic invasion and epithelial-mesenchymal transition (EMT) in oropharyngeal carcinomas

Background/Aim To examine the clinical correlation of p16 expression with Epithelial–Mesenchymal Transition (EMT) markers and lymphatic invasion in OPSCC patients in terms of clinical status at presentation, subsequent progression, and survival. Methods Tissue blocks of biopsy-proven Oropharyngeal Squamous Cell Carcinoma were subjected to Immunohistochemistry (IHC) for evaluating the expression of p16, e-cadherin, vimentin and podoplanin. This expression pattern was correlated with the demographic details, treatment response and survival patterns. Results 60 patients were finally available for evaluation in this study. Prevalence of HPV infection in our study was found to be 11.7%. E-cadherin expression was found in all HPV-associated patients whereas vimentin was not expressed in any of these. 71.4% patients had low Podoplanin expression and 85.7% had low lymphatic vessel count. Among the HPV- associated patients, 85.8% had T3-T4 stage, 100% had N2-N3 disease and 95% had stage IV disease. It was also found that p16-positive patients had significantly higher 1-year Overall survival (OS) (80%, p=0.045), higher 1-year Progression-free survival (PFS) (60%) and higher 1-year Locoregional recurrence-free survival (LRRFS) (75%) when compared to p16-negative patients. Conclusion Prevalence of HPV infection was found to be similar to that of previous studies conducted in India. As previous literature suggests, the HPV-positive patients in our study presented with advanced nodal disease at presentation and thereby, an advanced overall stage. Further follow-up of these patients including their treatment details, determination of possible prognostic markers, and evaluation of their survival parameters can be done which can help in modifying the existing treatment modalities as HPV-associated OPSCC are known to have better prognosis according to literature.

P0068 Distribution of α4β7 integrin and MAdCAM-1 expression according to the location of intestinal inflammation in a murine model of spontaneous chronic enterocolitis

Abstract Background The selective blockade of α4β7 integrin is a treatment for inflammatory bowel disease (IBD), with different efficacy according to the location of inflammation along the gastrointestinal tract. A more pronounced effect has been reported in patients with predominant colonic inflammation. It may be attributed to differences in the regional expression of adhesion molecules. This study aimed to investigate the expression of α4β7 integrin and mucosal addressin cell adhesion molecule-1 (MAdCAM-1) in colon, ileum, and mesentery, using IL-10 knockout (KO) mice as a model of spontaneous chronic enterocolitis. Methods At 20 weeks, the colon, ileum, and mesentery were extracted from IL-10KO and C57BL/6 wild-type (WT) mice. Protein levels of α4β7 integrin and MAdCAM-1 were determined by Western blot in the different tissues. Additionally, pro-inflammatory cytokines (TNF-α and IFN-γ) and epithelial adhesion molecules (Zonula Occludens-1, ZO-1; E-cadherin and Mucin 2, Muc2) expression in the ileum, as well as adiponectin expression in the mesentery, were analyzed using real-time PCR. Results Analysis demonstrated a significant increase in gene expression of the pro-inflammatory cytokines TNF-α and IFN-γ in the ileum of IL-10 KO mice compared with WT mice (p&amp;lt;0.01). In addition, IL-10KO mice exhibited reduced expression of ZO-1, E-cadherin, and Muc2 in the ileum relative to WT mice (p&amp;lt;0.001). IL-10 deficiency also led to a marked decrease in adiponectin expression in the mesentery (p&amp;lt;0.05). We then analyzed the protein level of MAdCAM-1 and α4β7 integrin according to anatomic region. Remarkably, the protein analysis revealed significantly higher levels of MAdCAM-1 (p&amp;lt;0.01) and α4β7 integrin (p&amp;lt;0.05) in the colon of IL-10 KO mice compared to WT. In the ileum, while MAdCAM-1 levels remained unchanged, α4β7 integrin expression was increased in IL-10KO mice (p&amp;lt;0.01). No significant differences in MAdCAM-1 or α4β7 integrin levels were observed in the mesentery between IL-10KO and WT mice Conclusion IL-10-deficient mice develop inflammation in the ileum, accompanied by a reduction in adiponectin production in the mesentery. In this setting, both α4β7 integrin and MAdCAM-1 showed increased expression in the colon, which was not observed in the ileum or mesentery, despite the inflammation. This differential regional expression pattern of adhesion molecules may explain the variation in the efficacy of anti-integrin therapies. Characterizing these patterns in patients with IBD could help identify those who would benefit most from these treatments.

P0080 Thalidomide-Carbomer Gel for Rectal Administration Alleviates DSS-Induced Colitis and fibrosis in Mouse by Targeting S1pr5 and Il1f9 (Il36γ)

Abstract Background Inflammatory bowel disease (IBD) is a chronic inflammatory condition of the digestive tract, including ulcerative colitis (UC) and Crohn's disease (CD). Thalidomide, an inhibitor of Tnf-α, is used in IBD treatment but is limited by systemic side effects when taken orally. This study aimed to develop a thalidomide-carbomer gel for rectal administration to reduce systemic side effects and explore new therapeutic targets. Methods An acute colitis model was established using DSS-induced wild-type C57 mice. Mice were treated with thalidomide-carbomer gel via rectal administration. Body weight, disease activity index (DAI), colon length, liver and spleen indices, and histological changes were evaluated. RNA-seq, qPCR, WB, and immunohistochemistry were performed to assess inflammation and fibrosis markers. In vitro, an LPS-induced inflammation model was constructed using NCM460 cells, and cell migration and inflammatory cytokine expression were analyzed following drug treatment. Results Thalidomide-carbomer gel significantly alleviated symptoms of DSS-induced colitis in mice, including reducing body weight loss, DAI score, colon shortening, and liver/spleen indices. Histological analysis showed decreased infiltration of inflammatory cells and reduced expression of Tnf-α, α-SMA, and Tgf-β. The gel also enhanced intestinal barrier function by increasing the expression of E-cadherin and Occludin. RNA-seq and qPCR analyses suggested that the therapeutic effect may involve the regulation of S1pr5 and Il1f9（Il36γ）pathways. Toxicity tests indicated no significant adverse effects on hematological or liver and kidney functions. In NCM460 cells, thalidomide inhibited LPS-induced cell migration and IL-6 release. Conclusion Thalidomide-carbomer gel, through the inhibition of Tnf-α, S1pr5, and Il1f9 (Il36γ), significantly improved DSS-induced colitis and fibrosis in mouse, showing promising therapeutic potential for IBD.

Processed Products of Aconitum soongaricum Stapf. Inhibit the Growth of Ovarian Cancer Cells In vivo via Regulating the PI3K/AKT Signal Pathway.

The alkaloids of songorine, aconitine, and benzoylaconitine, as the processed products of Aconitum soongaricum Stapf., can significantly inhibit the migration and invasion of ovarian cancer cells in vitro. Herein, we studied the in vivo role and mechanism of these natural products in processed A. soongaricum Stapf. A xenograft tumor model was constructed. Tumor volumes and weights were calculated. HE staining assessed the histopathological changes of tumors. Inflammatory factors were detected using ELISA. Gene and protein expressions of E-cadherin, N-cadherin, PIK3CA, and AKT1 proteins were measured using RT-qPCR and immunohistochemistry. Protein expressions of E-cadherin, N-cadherin, PIK3CA, AKT1, p-PIK3CA, and p- AKT1 proteins were detected using western blot analysis. Songorine, aconitine, and benzoylaconine significantly inhibited the growth of tumors as evidenced by decreased tumor volume and weight. The extent and scope of tumor cell necrosis were less in the songorine group compared to the vehicle group. Songorine, aconitine, and benzoylaconine significantly reduced IL-6, IL-1β, and TNF-α levels. Furthermore, songorine, aconitine, and benzoylecgonine induced down-regulation of N-cadherin and AKT1 mRNA in comparison to the vehicle group. Meanwhile, songorine, aconitine, and benzoylaconine also significantly reduced N-cadherin, p-PIK3CA, and p-AKT1 proteins, while upregulating E-cadherin protein expression in comparison to the vehicle group. These effects were further enhanced when combined with the PI3K inhibitor LY294002. Songorine, aconitine, and benzoylaconine may inhibit ovarian cancer growth in vivo by blocking the PI3K/AKT signaling pathway. Our findings may provide evidence for the clinical application of the processed products of Aconitum soongaricum Stapf. in ovarian cancer treatment.

P0117 Exploring the role and mechanism of leukotriene D4 in fistulizing Crohn's disease based on multi-omics research

Abstract Background Alterations in the gut microbiota and its metabolites are involved in the pathogenesis of Crohn's disease (CD). In a study targeting patients with different phenotypes of CD, we investigated the characteristics of the gut microbiota and metabolites between Crohn's disease with fistula and Crohn's disease without fistula. Methods We divided the research subjects into three groups, including 11 CD patients with fistulas, 9 CD patients without fistulas, and 5 healthy individuals (control group). Stool samples from patients were collected and sequenced. The fecal microflora was analyzed by 16S ribosomal RNA gene amplification sequence, and the metabolites were quantified by liquid chromatography tandem mass spectrometry (LC-MS/MS). Bioinformatics analysis was employed to explore the microbial and metabolite characteristics of CD fistulas. Then, we further verified through mouse experiments and cell experiments. Results Compared with healthy controls, CD patients mainly showed a reduction in the Firmicutes phylum and an increase in the Proteobacteria phylum. The reduction of these short-chain fatty acid-producing bacteria and the increase in harmful bacteria are likely important factors leading to CD. Compared with non-fistula patients, the beneficial bacteria such as the genus Romboutsia in fistula patients were significantly reduced. In addition, we also found structural disruption and functional dysregulation of microorganisms in fistula patients. Metabolomics analysis indicated that the increase in pro-inflammatory metabolites such as leukotriene D4 is a specific change in fistula. Spearman correlation analysis showed that the reduction of beneficial bacteria was significantly negatively correlated with the increase in leukotriene D4, suggesting that the reduction of beneficial bacteria may lead to an increase in leukotriene D4 and thus promote the formation of fistulas. In vitro experiments further demonstrated that leukotriene D4 can delay the healing of fistula in mice. Cell experiments also proved that leukotriene D4 can reduce the expression of E-cadherin and upregulate the expression of EMT-related genes (snail, vimentin). Transcriptome analysis revealed that leukotriene D4 may lead to the formation of fistulas by activating the AGE-RAGE signaling pathway in diabetic complications. Conclusion In CD patients, the reduction of beneficial bacteria may promote the upregulation of leukotriene D4, thus promoting the formation of fistula.

DOP107 Real-time intestinal barrier assessment by endocytoscopy and confocal laser endomicroscopy uniquely correlates with multiple barrier protein expression and reflects the gut-brain axis

Abstract Background The intestinal barrier is gaining recognition as a key indicator of mucosal healing in IBD. Nonetheless, its assessment remains challenging. This study evaluates advanced endoscopic tools for real-time gut barrier assessment, examining their correlation with automated epithelial and vascular barrier markers and exploring their potential to elucidate the gut-brain axis. Methods IBD patients undergoing endoscopic assessment or surveillance and healthy controls were included. The intestinal mucosa was assessed with advanced imaging, i.e. ultra-high magnification endocytoscopy (ECS) or confocal laser endomicroscopy (CLE), using newly developed scores for barrier evaluation.1,2 Barrier healing was defined as ECS ≤ 1 -ileum and colon- and CLE ≤ 4 -colon- or ≤ 3 -ileum-. Biopsies were collected from different bowel segments, including representative inflamed and non-inflamed areas. Epithelial and vascular barrier markers, including ZO-1, Claudin-2, E-cadherin, PV-1 and CD-31, were analysed through confocal microscopy, and their expression was automatically quantified using QuPath. The validated IBD disk, focusing on energy, sleep and emotional components, was employed to evaluate neuropsychological functional impairment and indirectly assess gut-brain interaction.3 A per-biopsy-based linear regression model was used for inferential analysis using R software. Results A cohort of 33 IBD patients (15 CD and 18 UC) and 2 healthy controls were recruited (65 biopsies; 3.8 X106 cells). 9/16 (56%) patients who underwent ECS in the colon and 6/9 (67%) in the ileum had ECS scores indicative of barrier healing. CLE assessment revealed features suggestive of barrier healing in 3/11 patients (27%) in the colon and 1/14 (7%) in the ileum. ECS and CLE sub- and total scores demonstrated unique correlation with epithelial and vascular barrier proteins. For the epithelial barrier, CLE total scores in colon and ileum and villi architecture at ECS significantly correlated with Claudin-2 mean expression (p&amp;lt;0.01). Regarding the vascular barrier, a significant correlation was found between ECS-assessed vascular architecture in ileum and colon and PV-1 mean expression (p&amp;lt;0.01). In contrast, CLE-assessed blood flow in the colon was significantly correlated with CD-31 expression (p=0.03). Interestingly, patients with higher levels of sleeping difficulties based on IBD disk showed higher Claudin-2 and lower E-cadherin expression, while those with altered CD-31 expression reported lower energy levels. Conclusion CLE and ECS may offer a unique ability for real-time assessment of barrier impairment in IBD, showing promises for targeting real-time barrier healing and unravelling the complexities of gut-brain axis. References 1Iacucci M, Jeffery L, Acharjee A, et al. Computer-Aided Imaging Analysis of Probe-Based Confocal Laser Endomicroscopy With Molecular Labeling and Gene Expression Identifies Markers of Response to Biological Therapy in IBD Patients: The Endo-Omics Study. Inflamm Bowel Dis. 2023;29(9):1409-1420. doi:10.1093/ibd/izac233 2Majumder S, Santacroce G, Maeda Y, et al. Endocytoscopy with automated multispectral intestinal barrier pathology imaging for assessment of deep healing to predict outcomes in ulcerative colitis. Gut. 2024;73(10):1603-1606. Published 2024 Sep 9. doi:10.1136/gutjnl-2024-332894 3Le Berre C, Flamant M, Bouguen G, et al. VALIDation of the IBD-Disk Instrument for Assessing Disability in Inflammatory Bowel Diseases in a French Cohort: The VALIDate Study. J Crohns Colitis. 2020;14(11):1512-1523. doi:10.1093/ecco-jcc/jjaa100