bluuebunny/crossref_metadata_2025_split huggingface dataset:
P0080 Thalidomide-Carbomer Gel for Rectal Administration Alleviates DSS-Induced Colitis and fibrosis in Mouse by Targeting S1pr5 and Il1f9 (Il36γ)
Authors: He X , Jiang L , Hu J , Yang H 
Published: January 2025

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.
P0199 Development of a high-throughput multiplex immunofluorescence and an AI-assisted image analysis tool to study T cells dynamics in pre-clinical rodent models of IBD
Authors: Preti M , Hofving T , Bretin A , Benonisson H , Gonzalez-King Garibotti H , Burgueno J , Borde A , Birrel M , Preite S , Piras A 
Published: January 2025

Abstract

Background
T cell therapies are emerging as a new approach to treat and cure patients suffering from immune diseases where T cell homeostasis is no longer maintained. For the evaluation of treatments in pre-clinical rodent models, tools are required that enable fast, observer-independent and high-resolution analysis of different T cells in tissue sections. Multiplex staining of tissue samples provides a valuable opportunity for comprehensive analysis of cellular dynamics with enhanced spatial resolution and tissue context compared to traditional staining and flow cytometry methods. We have developed a high-throughput multiplex immunofluorescence panel for detecting different T cell types and activation status in mouse models of inflammatory bowel disease (IBD). Moreover, we used an AI-assisted method to enhance image analysis, enabling automated, accurate, and quantification of complex cellular interactions.

Methods
For the development and optimization of the various stainings we used formalin-fixed paraffin embedded (FFPE) mouse colon sections from an efficacy study with murine polyclonal regulatory T cells (Tregs) in the T cell transfer mouse model of IBD, enabling access to sections with healthy versus diseased pathology, as well as samples where Tregs were transferred to reduce colon pathology. Multiplex staining was performed with antibodies (such as CD3, FOXP3, CD25, CD69, CTLA-4 and KI67) using a Leica BOND RX autostainer, by applying sequential primary antibody staining, followed by secondary HRP-conjugated antibodies and Opal-TSA chemistry. Whole slide images (WSI) were generated by a Phenoimager HT (Akoya) and analyzed using Visiopharm software, with DAPI+ nuclei detection via AI-based application.

Results
We successfully developed a multiplex panel that enables high throughput staining and spatial resolution analysis of over six markers. We were able to determine the activation status of different subtypes of T cells and their specific localization in the colon. We could quantify infiltration of activated effector T cells (Teffs) in both the mucosa and submucosa in colon samples with high pathological scores, as well as FOXP3+ cells in samples from mice treated with Tregs.

Conclusion
We developed an Opal-based multiplex immunofluorescence method to detect T cells in mouse colon samples, focusing on activation and proliferation markers. Additionally, an AI-assisted image analysis tool that provided fast, reproducible, and quantitative results with spatial resolution. This innovative multiplex method holds promise for adaptation to different mouse models of colitis and other rodent models related to immune diseases, furthering its potential impact in the area of T cell therapy research.
P0202 Modeling Inflammatory Bowel Disease in mice with “Humanized” immune system
Authors: Coupe B , Charton A , Placide F , Machet F 
Published: January 2025

Abstract

Background
Inflammatory bowel disease (IBD) is a chronic recurrent inflammatory disease with unknown etiology. Dextran sulfate sodium (DSS) induced colitis is a common preclinical mouse model in IBD research. DSS colitis involves activation of the submucosal immune system and can be used to study Ulcerative Colitis characteristics in different phases (acute, chronic and remission) and predict compound efficacy. However, optimizing translation to human is still needed and the use of mice with Human Immune System (HIS) provide a better model to study human immune system responses in vivo. HIS mice are created by transplanting human immune cells or their progenitor cells into highly immunodeficient recipient mouse hosts, thereby "humanizing" their immune systems. In this study we discussed colitis feature and the use of humanized mice model for testing therapeutics in preclinical studies

Methods
15-weeks-old female HIS mice were treated with 2.5% dextran sulfate sodium (DSS) in drinking water (ad.lib.) for 5 days to induce acute colitis. Colonic and systemic human cytokines were assessed by MSD plex assay. Human immune cell infiltration into the colon was measured by flow cytometry. Colonic histological damage was assessed on colon Swiss roll after H&E staining

Results
HIS mice challenged with DSS exhibited body weight loss and increase disease activity index (loose stool or diarrhea, rectal bleeding) compared to naive HIS mice. HIS-DSS mice show a strong human pro-inflammatory response, including elevated hIL-1B, hTNF, hIL-6 in plasma and colon as well as specific human cytokines (MCP-4, IL-8, IL-26, IL-37…). Colon microscopic analysis show loss of goblet cells, loss of crypt structure and increase of inflammatory cell infiltration at the level of mucosa and submucosa

Conclusion
These data show a significant increase of human pro-inflammatory cytokines after DSS intake as well as histologic score characteristic of Ulcerative Colitis. They also confirm that HIS mice represent a relevant model for the preclinical assessment of therapeutics in IBD.
P0221 The role of caffeine and its metabolites in the modulation of intestinal inflammation and epithelial barrier integrity
Authors: Kulokiene U , Inciuraite R , Butaite G , Salteniene V , Kiudelis V , Jonaitis L V , Kiudelis G , Kupcinskas J , Skieceviciene J 
Published: January 2025

Abstract

Background
Methylxanthine caffeine and its metabolites are widely consumed bioactive compounds that are known to influence various physiological processes 1. However, their impact on gut health, particularly in modulating inflammation and epithelial permeability, remains underexplored. In this study we aimed to investigate the effects of caffeine and its primary metabolites paraxanthine, theobromine and theophylline on inflammatory responses and barrier integrity in gut epithelial models, providing insights into their potential role in gastrointestinal health in the context of Inflammatory bowel disease (IBD).

Methods
Primary screening of different concentrations of caffeine and its metabolites paraxanthine, theobromine and theophylline was performed in Caco-2 cells. Selected metabolites were further tested on inflamed 3D intestinal epithelial organoids derived from ulcerative colitis patient (n = 1) and non-IBD individual (n = 1). Inflammation was induced for 24 h using TNF-α and IFN-γ cytokine mix. The effects on inflammation status and epithelial barrier integrity were tested through targeted gene expression analysis using RT-PCR, while the effects on generation of reactive oxygen species were investigated by Flow cytometry and DHE assay.

Results
Primary screening revealed that both caffeine and its metabolite paraxanthine affected the expression of genes encoding inflammatory cytokines (TNF-α, CXCL9; p &lt; 0.05) and tight junction proteins (OCLN, ZO-1, CLDN1; p &lt; 0.05) and were selected for further analysis in 3D intestinal organoids. Neither caffeine nor paraxanthine reduced the expression of inflammatory cytokine-encoding genes in inflamed intestinal organoids. However, both metabolites significantly restored the expression of tight junction protein-coding genes OCLN, ZO-1 and CLDN1 (p &lt; 0.05). A tendency of reduction in reactive oxygen species production (by 11 %) was also observed after exposure with paraxanthine.

Conclusion
Methylxanthine caffeine and its primary metabolite paraxanthine might have a positive effect on gut epithelial barrier function through restoration of the expression of tight junction protein-coding genes.
Study was funded by the European Union and the State Secretariat for Education, Research and Innovation (SERI) (project: miGut-Health, grant no. 101095470).

References
Reddy V. S. et al. (2024) Pharmacology of caffeine and its effects on the human body. European Journal of Medicinal Chemistry Reports, 10 (100138). https://doi.org/10.1016/j.ejmcr.2024.100138.
P0282 Fibroblast activation protein (FAP)-cleaved type III Collagen [C3F] is a potential marker for intestinal fibrosis in patients with Crohn’s Disease
Authors: Tsapanou Katranara T , Poulsen A , Sorokina Alexdóttir M , Buhl Riis L , Pehrsson M , Rasmussen J , Gögenur I , Burisch J , Karsdal M A , Bay-Jensen A C , Seidelin J B , Mortensen J H 
Published: January 2025

Abstract

Background
Crohn ‘s disease (CD) is characterized by chronic inflammation in the gut, where severe complications such as fibrotic strictures require surgical resection. Stricture development involves excessive extracellular matrix (ECM) deposition due to continuous activation of intestinal myofibroblasts, that further contribute to intestinal fibrogenesis. Emerging data suggests that stricture-related intestinal myofibroblasts overexpress a serine protease called fibroblast activation protein (FAP). Furthermore, type III collagen deposition is increased during fibrosis in all layers of the intestinal tract, and studies have shown elevated collagen degradation in serum from patients with CD. The present study aimed at evaluating the potential of FAP-cleaved type III collagen [C3F] as a serum marker for intestinal fibrosis in CD.

Methods
The cohort consisted of 49 patients with luminal CD and 62 patients with stenotic CD. Clinical assessment was conducted for all patients at baseline and 12 months post study initiation. For the luminal phenotype, endoscopy was performed at both time points. For the stenotic group, radiographical assessment was done prior to surgery and endoscopy was performed at 12 months. C3F was measured in serum at all timepoints for baseline, post-operative day (POD), 1, 3, 6 and 12 months. To compare C3F levels at baseline for luminal vs stenotic phenotype, Mann-Whitney U-test was conducted. For the stenotic group, mixed-effect analysis, with a Dunnett’s multiple comparison, was conducted to compare C3F levels for baseline vs POD and Spearman’s rank correlation was applied.

Results
C3F baseline levels were significantly higher in patients with stenotic CD compared to luminal CD (Figure 1). In the stenotic group, C3F was significantly increased in patients at POD compared to baseline (Figure 1). Furthermore, C3F showed a positive correlation with C-reactive protein (CRP) (ρ=0.28, p=0.006) and a significant negative correlation with disease duration (ρ= -0.257, p=0.048) for patients with the stenotic CD phenotype (Table 1).

Conclusion
C3F was elevated in patients with stenotic CD, with even higher levels after the surgical resection. The data suggests that C3F reflects fibroblast activity during fibrostenosis and tissue remodeling following surgery. The positive correlation with CRP supports the association with inflammation leading to collagen deposition and fibrosis. Interestingly, C3F shows negative correlation with disease duration suggesting that C3F could reflect fibrosis at early stages of stenotic CD. These findings highlight the potential use of C3F as a novel biomarker for intestinal fibrosis in CD.
P0291 Tailoring Crohn’s disease surveillance after ileocolic resection: the importance of inflammatory biomarkers
Authors: Ferreira A I , Lima Capela T , Xavier S , Arieira C , Cotter J 
Published: January 2025

Abstract

Background
Recurrence of Crohn’s disease (CD) after ileocolic resection is common. However, reliable non-invasive monitoring methods are lacking, for instance, the use of faecal calprotectin (FC) has showed inconsistent results in this subgroup of patients. The aim of this study was to evaluate clinical and biochemical factors associated with post-operative endoscopic recurrence in CD patients.

Methods
Retrospective single-center study including patients with CD who underwent ileocolonoscopy for CD surveillance, after ileocolic resection. The patients’ clinical status was evaluated through Harvey-Bradshaw index (HBI), with clinical remission considered as ≤ 4 points. Biochemical parameters included erythrocyte sedimentation rate (ESR), c-reactive protein (CRP) and FC. Clinical and biochemical parameters were evaluated in the 6 months prior to ileocolonoscopy. Post-operative endoscopic recurrence was defined as a Rutgeerts score ≥ i2b.

Results
A total of 81 patients and 121 ileocolonoscopies were included. Most patients were female (55.6%), with a mean age of 48±13 years. Regarding the Montreal classification, 37 patients had ileal disease (45.7%) and 44 ileocolonic disease (54.3%); 9 patients had nonstricturing, nonpenetrating disease (11.1%), 35 stricturing (43.2%) and 37 penetrating disease (45.7%). Nineteen patients had perianal disease (23.5%). The mean time between the surgery and the endoscopic evaluation was 135±79 months. Post-operative endoscopic recurrence was observed in 52 ileocolonoscopies (43.0%).
Symptomatic patients (HBI &gt; 4 points) were 11 times more likely to have post-operative endoscopic recurrence (OR 10.686, 95%CI 2.930-38.974, p&lt;0.001). Those with post-operative endoscopic recurrence had a significantly higher FC values (median 189 vs 69 µg/g, p&lt;0.001) and higher ESR values (median 14 vs 9 mm, p=0.002). In fact, patients with FC values ≥ 150 µg/g and those with ESR values &gt; 12 mm were 4 and 3 times more likely to have post-operative endoscopic recurrence (OR 3.654, 95%CI 1.605-8.316, p=0.002 and OR 3.059, 95%CI 1.397-6.696, p=0.005, respectively). The presence of post-operative endoscopic recurrence was not associated with CRP values (median 2.9 vs 2.9 mg/L, p=0.466), and neither with mean time between the surgery and the endoscopic evaluation (mean 126±83 vs 142±75 months, p=0.263).

Conclusion
The presence of symptoms (HBI &gt; 4 points), FC values ≥ 150 µg/g and ESR values &gt; 12 mm was associated with post-operative endoscopic recurrence in CD. Therefore, the HBI, FC and ESR values can be used to monitor CD patients after ileocolic resection, and prompt endoscopic evaluation should be performed in those with HBI &gt; 4 points, FC values ≥ 150 µg/g and/or ESR values &gt; 12 mm.
P0421 Clinical validity of a seroproteomic index of endoscopic activity in pediatric Crohn’s disease
Authors: Orlansky Meyer E , Dotson J , Saeed S , Zitomersky N , Focht G , Rabizadeh S , Sockolow R , Griffiths A , Navas-López V M , Sladek M , Baldassano R , Otley A , Spencer E , Ho J , Dervieux T , Turner D , Dubinsky M 
Published: January 2025

Abstract

Background
A seroproteomic index (i.e. the serum index) corresponds to 13 protein markers, including CRP, combined as a multianalyte assay with a score range 0-100 where higher score is associated with higher probability of endoscopic activity in adult CD. We aimed to evaluate its performances in pediatric CD.

Methods
Children with CD were enrolled from two separate cohorts. Cohort 1 enrolled CD from multiple centers in the United States (n=118) and cohort 2, was multinational (n=160). A simple endoscopic score for CD (SESCD) was collected (&lt;3 points indicate endoscopic healing) with a blood specimen. Clinical disease status was measured using the Harvey Bradshaw index (HBI, cohort 1) or pediatric CD activity index (PCDAI, cohort 2) (PCDAI&lt;10 or HBI&lt;5 indicate clinical remission). Fecal calprotectin (FC) was collected in 180 patients from both cohorts. Serum specimens were processed in Prometheus Laboratories, San Diego, CA. Statistical analysis consisted of linear regression analysis, calculation of receiver operating characteristics curves (ROC) with area under the curve (AUC), sensitivity, specificity, Odds ratio (OR).

Results
In the 278 enrolled children (median age 14 years [IQR: 12-16 years], 46% females), median SESCD was 5 points (IQR: 0-12 points, 63% with endoscopic active disease); median CRP was 2.5 mg/L (IQR: 0.3-8.4 mg/L), median FC was 592 µg/g (IQR: 135-1147 µg/g) and median serum index was 33 (IQR: 17-60 points). AUROC analysis yielded significantly better area under ROC curve (AUROC, 0.87±0.03) for FC than the serum index (0.78±0.04) and CRP (0.77±0.04) (p&lt;0.05) in distinguishing endoscopic active disease from endoscopic healing. There was a 9-fold higher likelihood of endoscopic active disease with serum index above 32 points (OR= 8.95 [95%CI: 5.02,15.96] p&lt;0.001) with 70% sensitivity and 79% specificity (PPV=85%; NPV=61% at 63% pretest) (Table).
In multivariate analysis EHI above 32 points (aOR= 3.00 [95%CI: 1.25, 4.76] p=0.043), active clinical disease (aOR= 2.01 [95%CI: 0.85, 4.76] p=0.114), FC&gt;250 µg/g (aOR= 5.61 [95%CI: 2.4,13.09] p&lt;0.01) and CRP above 3 mg/L (aOR=2.09 [95%CI: 0.9-4.85])(p=0.08) associated with active endoscopic disease) (Figure). In the subset with clinical remission (n=158), or CRP below 3 mg/L (n=136), or FC levels below 250 µg/g (n=116), EHI above 32 points was still associated with higher likelihood of endoscopic disease (OR=5.4 [95%CI: 2.6,11.1], OR= 6.8 [95%CI: 2.9,15.8], and OR=4.0 [95%CI: 1.3,12.8], respectively) (p&lt;0.05) (Figure).

Conclusion
These data support the association of the serum index with endoscopic outcome in pediatric CD patients.
P0423 Anastomotic leakage after ileoanal pouch surgery: risk factors and salvage
Authors: Moojen T , Vlug M , Visser E , Reijntjes M , Lange J , Bislenghi G , Carvello M , Warusavitarne J , Hompes R , Stassen L , Faiz O , Spinelli A , D’Hoore A , Bemelman W 
Published: January 2025

Abstract

Background
Chronic anastomotic leakage (AL) is the most common cause of pouch failure after restorative proctocolectomy with ileal pouch-anal anastomosis for Ulcerative Colitis. This study aims to investigate the factors associated with AL and successful salvage of leaking anastomoses after ileoanal pouch surgery.

Methods
This multicentre retrospective cohort study included patients ≥18 years old with Ulcerative Colitis or unclassified Inflammatory Bowel Disease who underwent ileoanal pouch surgery between 2016 and 2021 in six European centres, with &gt;12 months follow-up. The primary outcome was AL rate. Secondary outcomes included factors associated with the occurrence of AL, diagnosis, management and salvage of AL and stoma-free survival.

Results
411 patients were included, of which 13.6% (n=56) had a diagnosed AL with a significantly higher rate in low-volume centres. Of the 56 AL, 44 were diagnosed early and 12 late. Three-stage approach was associated with late diagnosis and treatment. AL were managed using various techniques. The overall AL salvage rate was 85.4%, 92% when diagnosed and treated early and 60% when diagnosed and treated late (p=0.010). Successful AL salvage was associated with long-term stoma-free status (p=0.002). The long-term stoma-free rate was 95.5% in patients with early diagnosed and treated leaks, but only 41.7% when diagnosed and treated late (p&lt;0.001).

Conclusion
Early diagnosis and treatment of AL diminishes the negative effect of AL after ileoanal pouch surgery. Pro-active anastomotic assessment is mandatory to enable early diagnosis and management especially in patients at increased risk of silent leaks such as three-staged patients.
P0429 Endocytoscopy in real-time assessment of histological and endoscopic activity in ulcerative colitis
Authors: Chaemsupaphan T , Ali M S , Paramsothy S , Leong R W 
Published: January 2025

Abstract

Background
With 500-fold magnification, endocytoscopy (CF-H290ECI; Olympus Medical Systems Corporation, Japan; ARTG 121183) is a novel advanced imaging technology enabling real-time imaging of the intestinal mucosa. Endoscopic-histological remission assessment in ulcerative colitis (UC) has the potential to become the standard of care. Endoscopists, however, are uninvolved in the assessment of histology. Endocytoscopy might improve workflow and efficiency in the evaluation of UC activity.

Methods
In this single-center, prospective cohort study at Concord Hospital, Sydney, UC participants consenting to colonoscopy and endocytoscopy for disease assessment were recruited. Methylene blue chromoendoscopy enhanced cellular imaging. Data collected were demographics, disease activity, Mayo endoscopic score (MES) and endocytoscopic activity based on crypt shape, inter-crypt distance, cellular infiltration, and extent of vascularization. Correlation against the MES and the Nancy histological index (NHI) was conducted and validation against the ErLangen Endocytoscopy in CoLiTis (ELECT) score, and the endocytoscopy score (ECSS) was performed using Kappa (κ) statistics and Spearman correlation coefficient (r). P value &lt;0.05 was deemed statistically significant. Duration and adverse events data were recorded.

Results
A total of 61 colonic bowel segments from 15 UC patients were evaluated (endoscopic improvement 69%, histological remission 67%) (Table 1). Endocytoscopy correlated significantly with MES using both the ECSS (κ=0.60, P&lt;0.001; r=0.78, P&lt;0.001) and the ELECT (κ=0.88, P&lt;0.001; r=0.81, P&lt;0.001) scoring systems. Endocytoscopy correlated with NHI using both the ECSS (κ=0.47, P&lt;0.001; r=0.69, P&lt;0.001) and the ELECT (κ=0.88, P&lt; 0.001; r=0.74, P&lt;0.001) systems. The ELECT score demonstrated superior diagnostic performance in determining histological remission with sensitivity, specificity, and AUROC of 100%, 85%, and 0.90 (95% CI: 0.78-1.00), versus 68.3%, 85%, and 0.88 (95% CI: 0.75-1.00) with ECSS (Figure 1). No serious adverse events were reported, apart from temporary urinary discoloration from methylene blue excretion.

Conclusion
Instant simultaneous endoscopic and histological assessment of UC is possible using endocytoscopy, which was safe, accurate and well-tolerated. The ELECT score reflects histology significantly better than the ECSS.