bluuebunny/crossref_metadata_2025_split huggingface dataset:
P0159 Effects of Transcranial Direct Current Stimulation (tDCS) on enteric nervous system (ENS) gene expression and intestinal barrier in chronic DSS colitis
Authors: Wiese J J , Manna S , Bubeck M , Patankar J , Siegmund B , Prüß M S , Hering N , Schumann M 
Published: January 2025

Abstract

Background
Chronic visceral pain is a clinical challenge in &gt;50% of Inflammatory Bowel Disease (IBD) cases. A randomized controlled trial (RCT) revealed that transcranial direct current stimulation (tDCS) can modulate brain activity and effectively alleviate chronic abdominal pain associated with IBD. To elucidate the underlying mechanism of tDCS, we implemented this technique in a mouse model of inflammatory, chronic DSS colitis.

Methods
The tDCS protocol (10 stimulations) was deduced from human tDCS studies. We administered an anodal direct current (0.5 mA, 20 min) to both primary motor cortices over two separate periods (5 consecutive days of stimulation per period separated by a two-day break). The study was controlled by a sham-tDCS group.
For determining mucosal barrier integrity murine intestinal mucosa was mounted to Ussing chambers. Histology was done (H&E staining of FFPE colon sections). Murine activity after tDCS was assessed by mouse tracking in home cages (RFID-chipped mice). Myenteric plexus (MP) cells from the murine terminal ileum, were isolated and cultivated for 10 days. Immunostaining and bulk-RNA sequencing (NovaSeq Xplus system, read length 150 bp, read depth 30 M copies/run) were performed.

Results
DSS caused an inflammation-driven thickening of the colon wall, resulting in an increased transmural resistance. This effect was ameliorated by tDCS (mean±SEM DSS+sham-tDCS: 72.2±8.2 Ω·cm²; DSS+tDCS: 48.1±2.1 Ω·cm²; MWU: p=0.007). Induction of a DSS colitis was associated with a reduced murine activity, which was partially recovered by tDCS (mean±SEM DSS+sham-tDCS: 70.7±15.2 rel.dist.U/day; DSS+tDCS: 205.9 ± 40.4 rel.dist.U/day; MWU: p=0.029).
Neuronal cell identity was confirmed in isolated murine MP cells by immunostaining (b3-tubulin, S100B). Bulk-RNAseq significantly separated tDCS- from sham-tDCS treated DSS colitis mice as revealed by PCA. Consecutively, 739 differentially expressed genes were identified in tDCS-treated colitis mice compared to sham-tDCS-treated DSS colitic mice (622 upregulated, 117 downregulated genes; log2FC≥1.5, p-adj&lt;0.05). KEGG and GO pathway analysis included neuroactive ligand-receptor interaction (p=7.94·10-3), gated channel activity (p=2.4·10-7), substrate-specific channel activity (p=1.99·10-7), leukocyte migration (p=3.99·10-7).

Conclusion
In colitis, tDCS has the capacity to modulate activity of DSS-treated mice and alters gene expression in MP cells. This reveals another layer of evidence for the impact of central pathways on the brain-gut-axis.

References
Neeb et al., Brain Stimulation, 2019
P0167 Circular extrachromosomal DNA (eccDNA): a potential diagnostic and prognostic tool to be introduced into the clinical practice of Inflammatory Bowel Disease
Authors: Foscarini E , Emoli V , Petito V , Di Vincenzo F , Masi L , Troisi S , Pane C , Piazzesi A , Turchini L , Pugliese D , Laterza L , Lopetuso L R , Gerovska D , Arauzo M , Putignani L , Regenberg B , Gasbarrini A , Scaldaferri F 
Published: January 2025

Abstract

Background
The diagnosis of Inflammatory Bowel Disease, which continues to increase year by year, is complex; these disorders can be confirmed by investigations such as fecal calprotectin, endoscopy, histology of biopsies, and CT scan; despite this, there is a lack of highly specific and minimally invasive techniques that allow not only diagnosis but also determination of the severity of the disease and its evolution. Given these challenges, the search for reliable and specific predictors that can direct the clinician toward personalized patient care is very important. In this context, can help extrachromosomal circular DNA (eccDNA) that are eukaryotic nuclear circle DNA molecules, independent from chromosomes, with a length ranging from a few hundred to a few thousand base pairs. In addition to its already known role in cancer, in IBD increased levels of eccDNA have been found compared with healthy controls. Circular DNA could represent a potential biomarker that can be isolated from peripheral blood or intestinal mucosa and is useful for making early diagnoses and assessing disease status.

Methods
We examined peripheral blood, collected by blood sampling . "Technical Tests" were performed in order to determine the right procedure for storing blood samples and optimize the results obtained in terms of eccDNA extraction . The samples will be divided into two groups, for the first group the procedure is to centrifuge the blood at 2000g for 15 minutes at different timepoints with or without storing the plasma at 4° C; for the second group the sample is centrifuged immediately after blood collection and the plasma is stored in the refrigerator at different timepoints [Table 1]. Then for both groups we proceed with eccDNA extraction by the "Circle-Seq" analysis.

Results
No significant differences are shown between the two groups in terms of the amount of eccDNA [Figure 1]. In particular, the non-immediate centrifugation and separation of plasma from whole blood does not negatively affect the extraction yield, highlighting that degradative factors are not present in whole blood. In addition, refrigerated storage for a longer period of time is a positive factor for eccDNA concentration, probably because in this way circular DNA found within microvesicles has time to be extruded, increasing its concentration in plasma.

Conclusion
Circulating eccDNAs could represent potential biomarkers to be detected in plasma by liquid biopsy, as the method is minimally invasive and easily reproducible in analytical laboratories, becoming a diagnostic and prognostic tool to be brought into clinical practice.

References
Fundings: European Union’s Horizon 2020 Research and Innovation Programme (899417)
P0168 Differential expression of enteric glial cell markers in inflamed and non-inflamed tissue of patients with Ulcerative Colitis during clinically active disease and remission
Authors: Beaudeau J , Katinios G , Biskou O , Lindqvist C M , Walter S A , Bednarska O , Söderholm J D , Salomon B , Halfvarson J , Schoultz I , Hjortswang H , Keita Å V 
Published: January 2025

Abstract

Background
Ulcerative colitis (UC), a major subtype of inflammatory bowel disease, is characterized by chronic inflammation of the colon and rectum. Enteric glial cells (EGC) play a crucial role in gut barrier maintenance and may contribute to UC pathophysiology. This study aims to investigate EGC and their associated proteins in patients during clinically active disease and in clinical remission following anti-inflammatory treatment.

Methods
Colonic or rectal biopsies were obtained at baseline from both inflamed and non-inflamed segments in 14 patients with clinically active UC initiating anti-inflammatory therapy and from the same segments when the patients were in clinical and endoscopic remission during follow-up. As control groups, colonic biopsies from 16 patients with irritable bowel syndrome (IBS)-Mixed and 16 healthy controls were included. Immunofluorescence staining assessed two EGC markers: the glial fibrillary acidic protein (GFAP+) and S100 calcium-binding protein B (S100β+). Relative estimates of inflammatory proteins in biopsies were analysed using Olink technology, In vitro, the EGC cell line CRL-2690 was exposed to interleukins (IL)-4, IL-6, and IL-18 at varying concentrations and durations, with GFAP expression analysed by western blot.

Results
In patients with UC, immunofluorescence analysis revealed significantly higher GFAP+ and S100β+ EGC counts in inflamed biopsies during active disease compared to macroscopically non-inflamed biopsies obtained during clinical and endoscopic remission. Compared to baseline biopsies from macroscopically non-inflamed mucosa, GFAP expression significantly decreased during follow-up (Figure 1), while S100β levels remained unaltered. Regardless of mucosal inflammatory status, patients with UC exhibited higher EGC counts than IBS-mixed patients and healthy controls. Our findings also showed a significant upregulation of EGC-associated pro-inflammatory proteins, such as IL-6, IL-8 and TNF in inflamed biopsies from patients with clinically active UC, compared to biopsies obtained when the patients were in remission as well as to healthy controls. Notably, these protein levels decreased during inactive UC, approaching levels observed in the healthy controls. In vitro, IL-6 upregulated GFAP expression dose- and time-dependently, while IL-4 and IL-18 induced expression in a less predictable pattern.

Conclusion
Elevated EGC counts, and pro-inflammatory proteins characterise inflamed mucosa in UC during clinical active disease highlighting their potential roles in UC related inflammation. These findings may suggest that EGC could be explored as a potential therapeutic target and may contribute to the discovery of novel biomarkers for disease monitoring.
P0169 Relapse of ulcerative colitis on dose reduction of tofacitinib is associated with markers of T-cell activation but not clinical features: a prospective, multicentre observational study
Authors: Honap S , Kim S , Whettlock E , Centritto A , Seah D , Baillie S , Luber R , Choong L M , Samaan M , Pavlidis P , Patel K , Dart R , John S , Irving P 
Published: January 2025

Abstract

Background
Tofacitinib (TOF) is an effective treatment for ulcerative colitis (UC). A relationship between dose and both response and side-effect profile is recognised for JAK inhibitors. Dose reduction from 10mg bd to 5mg bd is therefore associated with improved safety but also with a risk of relapse. The BRITE study aimed to identify clinical predictors and biomarkers of relapse in a cohort of patients with stable UC undergoing TOF dose reduction.

Methods
Steroid-free patients with stable UC responding to TOF 10mg bd for at least 8 weeks were followed from dose reduction to time of relapse or for one year. Relapse was defined by worsening modified Mayo score, including an increase in the rectal bleeding and endoscopic subscores of ≥ 1 point. Clinical and endoscopic assessment with biopsy was performed prior to dose reduction (baseline) and at end of trial. PBMCs were isolated at baseline and were cryopreserved. Flow cytometry analysis of phospho-STAT levels of STAT1, STAT3, STAT4 and STAT5 was performed in T cells from unstimulated and cytokine-stimulated baseline PBMC. In addition, CD4 T cells were isolated from PBMC and were stimulated with plate-bound anti-CD3 and anti-CD28 (TCR activation) for 5 days; cell supernatants were analysed for 17 CD4 T-helper (Th) cell effector cytokines using a 17-cytokine Luminex assay.

Results
50 patients were enrolled across 3 centres; baseline characteristics are displayed in table 1. 23 (46%) patients relapsed after a median duration of 23 weeks (IQR 9-45). Demographics, disease duration, prior advanced therapy exposure (including duration on TOF), and baseline clinical, biochemical (calprotectin/CRP), and histo-endoscopic disease severity were not associated with risk of relapse. In contrast, molecular differences in CD4 T-cell function were identified in relapsers and non-relapsers. pSTAT5 levels were significantly induced by IFNa in relapse patients (p&lt;0.05), but not in remission patients (Fig 1a), while canonical pSTAT5 activation by IL2 showed no significant differences between the two cohorts of patients. Furthermore, TCR activation of baseline CD4 T cells showed significantly higher expression of IL10 (p=0.01) in remission patients compared to relapse (Fig 1b). Higher levels of resting pSTAT1High expression that was refractory to IFNa stimulation in relapse patients but inducible in remission patients was also identified (p=&lt;0.01).

Conclusion
In this large prospective study of TOF dose reduction, in contrast to previous studies, no baseline clinical or endoscopic predictors of relapse were found. However, potentially clinically useful association with markers of T cell activation were identified and should be investigated further.
P0170 The presence of creeping fat in a TNBS-induced colitis model is associated with greater acute inflammation and bacterial translocation
Authors: Suau R , Clua L , Guasch M , Monfort-Ferré D , Boronat-Toscano A , Aquino M E , Matute K S , Cabrer M , Calafat M , Mañosa M , Domènech E , Jimeno M , Serena C , Bartolí R , Manyé J 
Published: January 2025

Abstract

Background
The hyperplasia of the mesenteric adipose tissue in Crohn’s disease, known as creeping fat, is associated with transmural lesions and the translocation of microbial antigens. In a rat model of TNBS-induced colitis, 50% of the animals presented adipose hyperplasia. Macroscopic evaluations showed that these animals had a more advanced progression of the disease, and colonoscopy could predict the presence of fat two days before euthanasia (L Clua et al, ECCO, 2024).

Methods
Colitis was induced using TNBS (n=10; half female), with the animals being sacrificed after five days. Tissue from the lesion along with mesenteric adipose tissue was obtained for histopathological staining and evaluation (haematoxylin-eosin, Masson’s trichrome, and Gram). Samples of intestinal mucosa and mesenteric fat were preserved in RNAlater to assess the expression of inflammatory markers by qRT-PCR. Results were statistically compared between animals with and without creeping fat, including sex.

Results
In histopathological analysis, using a validated scoring system, animals with creeping fat showed significantly greater severity of acute inflammation at the transmural level, with an increase in cryptitis and a more pronounced and extensive fibrosis pattern. Additionally, colonies of Gram-positive cocci were detected in all animals with creeping fat. In contrast, animals without fat hyperplasia exhibited appreciable adipogenesis in only two individuals, with an average of 105 adipocytes per field, compared to 185 in animals with creeping fat. qRT-PCR revealed higher expression of Il1b, Il6, Il10, Nod2, and Tlr2 in the intestinal mucosa and hyperplastic mesenteric fat of animals with creeping fat. A significant increase in Tnfa was also observed in hyperplastic adipose tissue, while the expression of Tgfbr1 was higher in the intestinal mucosa of animals without creeping fat. No differences were found between sexes.

Conclusion
The results indicated a more intense acute inflammatory and fibrotic component in animals with creeping fat, consistent with the macroscopic observations. The hyperplasia and inflammation observed in the mesenteric adipose tissue may be provoked by bacterial translocation, whereas animals without hyperplasia present a component influenced by TGF-β1 activity that could help counteract the inflammation.

References
L Clua, R Suau, M Guasch, D Monfort-Ferré, I Vañó, C Serena, E Domènech, R Bartolí, J Manyé, P022 Development of a novel experimental model for studying creeping fat in Crohn’s Disease, Journal of Crohn’s and Colitis, Volume 18, Issue Supplement_1, January 2024, Pages i275–i276, https://doi.org/10.1093/ecco-jcc/jjad212.0152
P0171 IL-36 signaling as a drug target in Crohn’s disease patients with IL36RN mutations
Authors: Hecker J , Plattner C , Löscher B S , Saurenbach J , Letizia M , Rieder D , Freise I , Koop K , Neufert C , Kunkel D , Schütz A , Becker C , Atreya R , Trajanoski Z , Franke A , Sonnenberg E , Siegmund B , Weidinger C ,   The TRR241 IBDome Consortium
Published: January 2025

Abstract

Background
The IL-36 signaling pathway has recently been identified as a key regulator of intestinal homeostasis and inflammation. However, the role of mutations in the IL-36R signaling pathway in the pathogenesis of inflammatory bowel disease remains unclear.

Methods
Mutations in the IL-36 receptor antagonist (IL-36RA, IL36RN) were identified in a cohort of 86 ulcerative colitis patients, 244 Crohn’s disease patients and 45 non-inflamed controls by whole exome sequencing followed by targeted Sanger sequencing. In-depth immune cell profiling of one IL36RN-mutated patient was performed using mass cytometry and multiplexed immunoassays. Overexpression experiments and functional assays characterized identified IL36RN mutations. Clinical and molecular responses of one IL36RN-mutated patient to a combined treatment with the IL-36R-blocking antibody spesolimab and the TNF-blocker certolizumab-pegol were assessed by fecal calprotectin, endoscopy, multiplexed immunoassays and mass cytometry.

Results
We identified four Crohn’s disease patients with heterozygous missense mutations in IL36RN. Experimental overexpression and functional assays demonstrated that two identified mutations resulted in a reduced protein expression of IL-36RA. In-depth immune profiling of one IL36RN-mutated patient revealed an increased response of PBMCs to IL-36 stimulation and elevated serum levels of IL-36-regulated cytokines. Administration of the IL-36R-blocking antibody spesolimab in combination with certolizumab-pegol over a period of 22 weeks to this patient resulted in a reduction of intestinal inflammation and alterations in immune cell composition and function.

Conclusion
Our findings indicate that pathogenic IL36RN mutations may contribute to the pathogenesis of Crohn’s disease in a subset of patients and that inhibiting IL-36 signaling in combination with TNF-blockade could offer a personalized therapeutic approach for these patients.

Disclaimer
Spesolimab was made available based on an out-of-scope request. Boehringer Ingelheim was given the opportunity to review this abstract as a courtesy. Boehringer Ingelheim had no role in the design, analysis or interpretation of the results in this study.
P0173 The impact of dietary composition on metallothionein gene expression in the intestinal epithelium by exclusive enteral nutrition
Authors: Curry D , Lemons J , Friedman E , Li Y , Hao F , Li H , Liu L , Patterson A , Henao-Mejia J , Lewis J , Wu G 
Published: January 2025

Abstract

Background
The mechanism by which exclusive enteral nutrition (EEN) is effective in the treatment of Crohn’s disease remains elusive. Epidemiologic studies have shown that a substantial proportion of patients with Crohn’s disease are deficient in micronutrients such as zinc, which has been shown to play an important role in the integrity of intestinal epithelial barrier function. Zinc plays a critical role in the transcriptional regulation of metallothioneins (MT), cysteine rich binding proteins that regulate both epithelial and immune function where they are generally believed to play a beneficial role in reducing disease.

Methods
Transcriptomic analyses were performed by both bulk RNAseq as well as GeoMx spatial transcriptomics. Enzymatic food digests using both bile acids and pancreatic enzymes, analyzed by Proton NMR metabolomics, were used to generate cell culture media for studies in intestinal epithelial cells.

Results
RNAseq of rectal biopsies obtained from healthy human subjects consuming EEN (Modulen-IBD, Nestlé Health Science) vs. an omnivore diet revealed a very specific induction of genes involved in mineral transport, namely MTs such as MT1G and MT2A. Spatial transcriptomics revealed that the induction of MTs was specific to the colonic epithelium. To develop a cellular model to characterize the mechanism underlying this response, we investigated the impact of animal protein, vegetable, and EEN based foods on the transcriptome of intestinal epithelial cells in culture by both RNAseq and targeted quantitative RT-PCR. This model involved the development of a physiologically relevant modality to digest whole food to generate serum-free cell culture media that was characterized using NMR-based metabolomics. Animal protein and EEN diets induced the expression of specific MTs whereas vegetable diets reduced their expression. We also show that zinc induced MT gene expression in intestinal epithelial cells whereas phytic acid, a phytochemical with known anti-nutrient properties that chelates zinc, reduces the ability of protein-based foods and EEN to induce MT gene expression.

Conclusion
The consumption of EEN leads to a very specific transcriptomic signature of MT gene expression in the rectal epithelium of humans that can be reproduced in cell culture by exposure to both animal protein-based foods and EEN. By contrast, vegetable-based foods inhibit MT gene expression possibly through the chelation of zinc. These results are consistent with the notion that the exclusion of plant-based foods through the consumption of EEN may have beneficial effects in the treatment of Crohn’s disease by reducing exposure to anti-nutrient phytochemicals that reduce the absorption of beneficial micronutrients such as zinc.
P0175 A new blocker of endothelial dysfunction ameliorates colitis independently of IL-10 by regulating both barrier function and the recruitment of immune cells
Authors: Park I S , Kim J H , Kim Y W , Kim D , Shin Y , Kim K B , Kim T I , Kim Research professor S W , Kwon Y G , Cheon J H 
Published: January 2025

Abstract

Background
Inflammatory bowel diseases (IBDs) like Crohn’s disease (CD) and ulcerative colitis (UC) are chronic gastrointestinal disease with complex pathogenesis, leading to a lack of effective treatments. The efficacy of CU104, a new blocker of endothelial dysfunction,1 in other IBD models with a CD-like phenotype2 and its specific mechanism of action in colitis are still unknown.

Methods
To evaluate the therapeutic potential of CU104, we induced colitis using dextran sodium sulfate (DSS) in IL-10 knockout mice3 and assessed its effectiveness. Additionally, we used Caco-2, HCT-116, and HT-29 cells to investigate its impact on intestinal barrier function, inflammatory signal pathway, actin dynamics,4 and gene expression through FITC-dextran permeability, trans-epithelial electrical resistance (TEER), reporter gene, gene expression profiling, and immune assays.

Results
CU104 demonstrated therapeutic efficacy in DSS-induced colitis in IL-10 KO mice by potently suppressing innate immune activation, vascular and intestinal barrier dysfunction, and immune cell recruitment. Mechanistically, CU104 inhibited the activation of nuclear factor kappa-light-chain-enhancer of activated B cell (NF-κB) and interferon regulatory factor (IRF),5 as well as the ezrin/radixin/moesin (ERM) signaling pathways6 in vitro and in vivo, which play critical roles in maintaining barrier integrity and regulating immune cell recruitment during inflammation by regulating actin dynamics.

Conclusion
Our findings suggest that CU104, a pseudo-sugar derivative of cholesterol that regulates actin dynamics and inflammatory signaling, could be a promising new treatment for CD along with UC.

References
1.Kim, Y. S. et al. CU06-1004 Alleviates Experimental Colitis by Modulating Colonic Vessel Dysfunction. Front Pharmacol 11, 571266 (2020).
2.Katsandegwaza, B., Horsnell, W. & Smith, K. Inflammatory Bowel Disease: A Review of Pre-Clinical Murine Models of Human Disease. Int J Mol Sci 23 (2022).
3.Wei, H. X., Wang, B. & Li, B. IL-10 and IL-22 in Mucosal Immunity: Driving Protection and Pathology. Front Immunol 11, 1315 (2020).
4.Kang, S. et al. TRIM40 is a pathogenic driver of inflammatory bowel disease subverting intestinal barrier integrity. Nat Commun 14, 700 (2023).
5.Li, J. Y. et al. IRF/Type I IFN signaling serves as a valuable therapeutic target in the pathogenesis of inflammatory bowel disease. Int Immunopharmacol 92, 107350 (2021).
6.Heemskerk, N. et al. F-actin-rich contractile endothelial pores prevent vascular leakage during leukocyte diapedesis through local RhoA signalling. Nat Commun 7, 10493 (2016).