bluuebunny/crossref_metadata_2025 huggingface dataset:
P0136 The role of FDG PET/CT in the and identification of fibrosis related to Crohn’s disease
Authors: Élthes Z , Sándor K , Galuska L , Farkas B , Pető S , Palatka K
Published: January 2025

Background

There is a constant search for non-invasive evaluation methods for the identification of fibrosis in patients treated for Crohn’s disease, since this condition can severely impact patients’ quality of life in the long run. While PET/CT is mostly used in oncological evaluation, it was previously proved that it can be useful in the evaluation of inflammation associated with Crohn’s disease activity, since segmental FDG uptake is proportional with immune cell infiltration of the bowel. However, it’s role in the detection of fibrosis needs further evaluation.

Our aim was to prospectively evaluate the prognostic value of FDG/PET CT in the identification of fibrosis related to Crohn’s disease regarding global PET scores compared to other imaging techniques (i.e. MRI, CT), simple endoscopic score (SES-CD), CD Activity Index (CDAI) intraoperative findings and histological evaluation of resected bowel segment.

Methods

We prospectively evaluated our patients from in and outpatient care between October 2021 and October 2024 who underwent bowel segment resection due to stenosis that was identified at a follow-up colonoscopy.

On PET-CT, we scored the FDG uptake in the small intestine and the 4 colon segments. Regions with increased FDG uptake were segmented with metabolically active volume (MAV), uncorrected maximal standardized uptake value (SUVmax), related to the SUVmax of the liver (as a reference for normal tissue activity) and uncorrected total lesion glycolysis (TLG).

Results

A total of 17 patients were selected, 11 (65 %) males and 6 (35 %) females, with a median age of 43 years (29-44). Ten patients (59 %) had Crohn’s disease affecting the small and large bowel, while in 3 (18 %) patients it affected only the small intestines and in 4 cases (23 %) it was localized solely on the large intestines. Seven (41 %) patients were bio naive, while 10 patients (59 %) received at least one biological therapy since their diagnosis. Five patients had a history of previous resection, from which three underwent endoscopic dilatation as well before the current surgery. In active disease, PET was more informative than endoscopy to access the extent of the inflammation, and small intestine involvement.

FDG PET-CT proved to be statistically more efficient in the identification of affected bowel segments than MRI or colonoscopy. However, we could not find an association between FDG PET-CT scores and the presence of fibrosis in our patients.

Conclusion

FDG PET-CT is a non-invasive complementary method for the evaluation of the affected bowel segments in CD, however its role in the identification of fibrosis is still not evident. This needs further investigation and a larger number of patients.
P0140 Estrogen Related Receptor Alpha Contributes to Intestinal Homeostasis by Modulating the Gut Microbiota and activating autophagy
Authors: Lee J Y , Kim S , Shin S G , Kim J K , Silwal P , Kim Y J , Shin N R , Kim P S , Won M , Lee S H , Kim S Y , Sasai M , Yamamoto M , Kim J M , Jo E K , Bae J W
Published: January 2025

Background

The gut microbiota plays a pivotal role in maintaining intestinal homeostasis and mitigating inflammation. The orphan nuclear receptor ESRRA (estrogen-related receptor alpha) has been identified as a critical regulator of mitochondrial function and autophagy, yet its role in gut microbiota modulation and intestinal health remains unclear.

Methods

We investigated role of ESRRA in intestinal homeostasis using Esrra-deficient mice in a dextran sodium sulfate (DSS)-induced colitis model. Colitis severity is investigated through survival rate and body weight. Gut barrier function was evaluated using FITC-dextran. Autophagic flux was evaluated using qPCR and histological methods in HT-29 cell. ESRRA expression was also assessed in human ulcerative colitis (UC) tissue samples. Gut microbiota composition was analyzed through 16S rRNA sequencing using QIIME2. Differential abundant taxa were identified using LEfSe analysis.

Results

Esrra-deficient mice exhibited heightened susceptibility to DSS-induced colitis, with body weight loss, disrupted gut barrier integrity, and elevated inflammation (Figure 1A-E). Autophagy regulators and markers Tfeb, Vps11, and Uvrag were significantly decreased in Esrra-deficient mice after DSS treatment (Figure 1F). Indeed, the mCherry-EGFP-LC3B reporter system showed secretion of autophagosome was not properly activated in Esrra-deficient cells (Figure 1G). ESRRA was also decreased in UC patients from GSE9452 and GSE11223 studies (Figure 1H). Immunohistochemical staining showed ESRRA was significantly reduced in UC colon tissues and gene set enrichment analysis showed autophagy is correlated with ESRRA expression (Figure 1I, J). Esrra-deficient mice also displayed altered gut microbiota composition, characterized by increased microbial diversity (Figure 2A-D). Unweighted UniFrac distance-based PCoA showed a significant cluster of each group (Figure 2E). LEfSe analysis showed Alloprevotella was significantly decreased in Esrra-deficient mice before/after DSS treatment (Figure 2F, H). Barnesiella and Anaserofustis were significantly decreased by DSS treatment in WT and KO mice (Figure 2G, I).

Conclusion

This study, published in the journal Autophagy 1, reveals that ESRRA is a vital regulator of intestinal homeostasis, coordinating autophagic activity and gut microbiota to protect against colitis. Targeting ESRRA or its downstream pathways could offer novel therapeutic strategies for inflammatory bowel diseases.

References

1.Kim S, Lee JY, Shin SG, Kim JK, Silwal P, Kim YJ, et al. ESRRA (estrogen related receptor alpha) is a critical regulator of intestinal homeostasis through activation of autophagic flux via gut microbiota. Autophagy 2021; 17:2856-75.
P0141 Estrogen receptor β alleviates colitis in intestinal epithelial cells by activating HIF-1α and ATG-9a mediated autophagy
Authors: Li J , Chen Y , Zhu L
Published: January 2025

Background

Ulcerative colitis (UC) manifests as excessive inflammation and compromised integrity of the intestinal epithelial barrier. Estrogen receptor β (ERβ) exerts anti-inflammatory effects in the colon. However, the mechanism remains unclear. This study aims to explore the role and mechanism of ERβ in alleviating colitis, and provide a novel therapeutic target for UC.

Methods

The level of ERβ and autophagy in the inflammatory and non-inflammatory colons of patients with UC were assessed. Mice with dextran sulfate sodium (DSS)-induced colitis were used to establish an in vivo model. The body weight change, colon length, disease activity index (DAI) and histological score of mice in each group were compared. In vitro, HT-29 cell inflammation was induced by lipopolysaccharide (LPS). ERB041, an ERβ specific agonist, was administered to DSS-induced mice and LPS-induced HT-29 cells. Autophagy in vivo and in vitro was evaluated by western blot, transmission electron microscope, and immunofluorescence. Co-immunoprecipitation, chromatin immunoprecipitation, and dual-luciferase reporter assay were conducted to further delineate the mechanism between ERβ and autophagy.

Results

Compared with non-inflammatory mucosa, the expression of ERβ in the inflammatory mucosa in patients with UC was down-regulated, with autophagy inhibited significantly. ERβ activation attenuated the intestinal epithelial barrier disruption, decreased the level of pro-inflammatory cytokines, and significantly attenuated DSS-induced experimental colitis. Mechanically, ERB041 intervention activated autophagy, which was manifested as up-regulation of LC3BII expression, decreased p62 level and increased number of autolysosomes. ERβ interacted with hypoxia-inducible factor 1α (HIF-1α) and activated autophagy. Furthermore, HIF-1α bound to the promoter region of ATG- 9a, and positively regulates its transcription.

Conclusion

ERβ exerts anti-inflammatory effect by up-regulating HIF-1α and activating ATG-9a mediated autophagy in intestinal epithelial cells, and may be a promising therapeutic target for UC.
P0142 CD47-SIRPα Contributes to the Microenvironment of Stricturing Crohn's Disease by Mediating the Crosstalk between Macrophages and Stromal cells: “Eat Me” or “Don't Eat Me”
Authors: Zhao X , Wang D , Sun S , Zhang Y , Zou D
Published: January 2025

Background

Intestinal fibrosis is a common complication of Crohn’s disease (CD), but no anti-fibrotic therapy is currently available. Collagen deposition, the pathological characteristics of intestinal fibrosis, is driven by an imbalance between extracellular matrix (ECM) production and degradation, which is controlled by the crosstalk between stromal cells and macrophage phagocytosis. The underlying mechanism of macrophage phagocytosis is the interplay between “eat me” signals and “don’t eat me”, among which CD47-SIRPa is the dominant pair. This study aims to investigate the role and therapeutic potential of CD47-SIRPa in intestinal fibrosis.

Methods

Thirty CD patients who underwent surgery due to fibrosis-induced intestinal obstruction were included in this study and the intestinal tissues were collected from both fibrotic and normal sites. Col1a2-CreERT2; Cd47 flox mice and Lyz2-Cre; SIRPα flox mice were used to conditionally ablate CD47/SIRPα. Mice intestinal fibrosis was induced by chronic dextrane sodium sulfate (DSS) and anti-CD47/anti-SIRPα antibody was treated intraperitoneally. Human and mice tissues were applied for single-cell RNA sequencing, flow cytometry, pathological evaluation, immunofluorescence staining, qPCR and in vitro experiment. Smart-seq were performed to detect the functional change of mice macrophage.

Results

Single-cell RNA sequencing and flow cytometry showed the proportion of CD47+ stromal cells and SIRPα+ macrophages significantly increased in intestinal fibrosis (Figure 1A). CD47+ stromal cells were in senescence-associated secretory phenotype (SASP), while macrophages also acquired a pro-fibrotic immature phenotype (all p<0.05). Moreover, we found the elevation of macrophage SIRPα expression was caused by CD47+ stromal cells and they extensively co-located in fibrotic lesions to form a pathogenic niche(Figure 1B). Conditionally ablation of CD47 in stromal cell or SIRPα in macrophage could significantly reduce collagen deposition(Figure 1C). Meanwhile, both anti-CD47 and anti-SIRPα treatment significantly attenuated mice intestinal fibrosis compared to the control.(Figure 1D) Mice intestinal macrophages acquired an anti-fibrotic phenotype and regained the capacity of phagocytosis by upregulating Clec7a and MMP10 after antibody treatment (all p<0.05) (Figure 1E). Furthermore, under the same total treatment dose, initiating antibody intervention at the earliest stage could achieve the best prognostic efficacy, highlighting its potential to be a therapeutic and preventive strategy against intestinal fibrosis (Figure 1F).

Conclusion

CD47-SIRPa mediated macrophage-stromal cell crosstalk significantly contributed to structuring CD and may be a novel therapeutic target for intestinal fibrosis.
P0145 Role of PPARg on anti-inflammatory effects mediated by AhR ligands in a Caco-2/THP-1 in vitro “inflamed gut” model
Authors: Oliveira Da Rocha G H , Müller C , Przybylski-Wartner S , Schaller H , Riemschneider S , Lehmann J
Published: January 2025

Background

Inflammatory Bowel Diseases (IBD) lack therapeutic options capable of inducing long-term remission in a cost-efficient manner. Repurposing strategies have allowed already established receptors and ligands to be tested as alternative pharmacotherapy for treatment of IBD. Agonists of the aryl hydrocarbon receptor (AhR), a ligand-dependent transcription factor traditionally associated with xenobiotic toxicity, have been explored as potential anti-inflammatory drugs to treat IBD, but specific mechanisms have yet to be elucidated. Like AhR, the peroxisome proliferator-activated receptor gamma (PPARg), responsible for control of lipid metabolism, has been evidenced as a key player in control of inflammation. In this study, we have explored a hypothetical crosstalk between PPARg and AhR in attenuating intestinal inflammation.

Methods

Bioinformatics analyses of a GEO RNAseq dataset of gut macrophages from IBD patients were used to assess correlations between PPARg and AhR. In vitro, macrophage-differentiated THP-1 cells and Caco-2 gut epithelial cells were seeded in Transwell® inserts to generate a gut-like co-culture model. Cells were treated with 100 ng/mL lipopolysaccharide (LPS) for 24 h in order to mimic a phenotype of intestinal inflammation, and also with combinations of AhR/PPARg agonists and antagonists at concentrations of 10 µM each. Proprietary AhR agonists were also tested. AhR translocation and cytokine secretion in macrophages as well as monolayer integrity and expression of tight junction proteins (e.g., claudin-2) in epithelial cells were assessed.

Results

Principal component analysis and Pearson correlation tests of the RNAseq dataset revealed that PPARg and AhR correlate with each other during intestinal homeostasis, but their expression patterns diverge during gut inflammation. In our co-culture model, blocking PPARg in THP-1 macrophages with a specific antagonist (GW9662) prevented AhR agonist 6-formylindolo[3,2-b]carbazole (FICZ) from inducing AhR nuclear translocation and secreting interleukin (IL)-6. In Caco-2 gut epithelial cells, GW9662 blocked the anti-inflammatory effects of FICZ on monolayer disruption and claudin-2 expression. These effects of FICZ were confirmed to be AhR-dependent, as they were prevented under treatment with the AhR antagonist CH223191. Proprietary AhR ligands also inhibited IL-6 secretion, which was shown to be the most dominant parameter in terms of converging anti-inflammatory effects of both PPARg and AhR.

Conclusion

Our data points to a pivotal cross-talk between AhR and PPARg, where PPARg is likely recruited downstream of AhR activation and contributes to the anti-inflammatory effects of FICZ, especially inhibition of IL-6 secretion.
P0151 Multi-modal single cell sequencing identifies CCR9 expression in lamina propria with highest protein expression on CD4 tissue resident memory cells in the inflamed ileum in Crohn’s disease
Authors: Hamilton R , Collin J , Keith B , Gehrmann U , Lindkvist B , Lundin S , Eberhardson M , Marks D , Platt A , Muthas D , Corridoni D , Bouma G
Published: January 2025

Background

Crohn’s Disease (CD) is a chronic condition that can impact any part of the gastrointestinal tract. The sustained inflammation is driven by the continuous influx of inflammatory leukocytes into the gut mucosa, which is regulated through chemokine gradients and adhesion molecules, including integrins. In the gut, the migration of inflammatory cells to specific segments is thought to be mediated by selected integrins and/or chemokine axes. The CCL25/CCR9 axis has been identified as a key pathway for immune cell migration into small bowel segments, such as the ileum. In this study, CITE-seq and spatial transcriptomic analysis was used to integrate proteomic and transcriptomic profiles to characterise CCR9+ immune cells in the ileum compared to the colon.

Methods

Biopsies were obtained via ileocolonoscopy from macroscopically inflamed and normal mucosa with appropriate consent from healthy volunteers and IBD patients visiting the Department of Gastroenterology at the Sahlgrenska Hospital. Biopsies were frozen in Cryostor CS10 or formalin fixed and paraffin embedded. Cells from frozen biopsies were sorted, CD45+ cells stained with CCR9 and Totalseq-C human universal antibody cocktail, followed by amplification, separation of antibody-derived DNA from mRNA derived cDNA, library construction and sequencing of both libraries. For spatial transcriptomics, tissue sections were deparaffinised, stained with H&E and decrosslinked before human whole transcriptome probe hybridisation, ligation, extension, library construction and sequencing. Cells were annotated using a transfer learning approach combining several reference single cell datasets including detailed CD4+ and CD8+ T cell datasets and utilising both antibody data and mRNA transcriptomic data.

Results

CCR9 positive cells appeared scattered throughout the lamina propria and expression at mRNA level was enriched in the ileum compared to the colon with about 10% of CD4 and 17% of CD8 T cells expressing CCR9. Interestingly at protein level, expression was more restricted with the highest expression on cells that exhibited proteomic and transcriptomic features associated with resident CD4 memory T cells (Trm). This population almost exclusively (>90%) consisted of CCR9 positive cells and their proteomic and transcriptomic profile consistent with the recently described effector and innate-like phenotype of Trm cells that is thought to contribute to Crohn’s disease pathology.

Conclusion

These data may point to CCR9 positive cells, and in particular Trm cells, in the ileum as important pathological cells in Crohn’s disease and provide further support for depleting CCR9 cells as a potential therapeutic target for the treatment of ileal Crohn’s disease.
P0154 Adenosine A2B receptor signaling induces mucin expression in the intestinal epithelium
Authors: Jäverfelt S , O’Connell L V , Clare K , Aherne C M
Published: January 2025

Background

Loss of mucosal barrier integrity and inappropriate perpetuation of the inflammatory response are key features of ulcerative colitis (UC). Increased permeability and disruption of the mucus layer lining the colonic intestinal epithelium is observed prior to the onset of colitis in UC and murine colitis models. Therefore, understanding how the integrity of the mucus layer is maintained during health and disease may lead to novel therapeutic approaches for UC.

The major constituents of the mucus layer are mucins which are secreted by goblet cells in the intestinal epithelium. MUC2 is the major mucin within the ileum and colon, and MUC5AC has recently been demonstrated to be protective in murine colitis. Extracellular adenosine signaling can mediate mucin hypersecretion in asthma and other inflammatory respiratory conditions. Studies suggest that signaling via the Adora2b adenosine receptor (A2BAR) is protective in acute gastrointestinal inflammation such as colitis, although the mechanism by which this occurs is not known. We hypothesized that A2BAR signaling induces MUC2 and MUC5AC expression in the intestinal epithelium.

Methods

We are combining human epithelial cell lines and mouse models to investigate the A2B-mediated effects on MUC2 and MUC5AC in the intestine during homeostasis and injury. MUC2 and MUC5AC mRNA expression were quantified by Taqman RT-PCR and MUC5AC protein expression by immunofluorescent stainings. To investigate A2B-mediated effects on mucus thickness, growth and penetrability, we use an ex-vivo method where the intestinal tissue is mounted in a horizontal perfusion system equipped with temperature control. Mucus thickness, expansion and penetrability to bacteria-sized beads are recorded before and after basolateral and apical stimulation with a selective A2B during a total of 60 minutes.

Results

Treatment of human intestinal epithelial cells with an A2B selective receptor agonist induced upregulation of MUC2 and MUC5AC mRNA, which was abolished by receptor blockade and inhibition of PKA. Similarly, stimulation of the A2B-receptor with the selective A2B agonist increased MUC5AC promotor activity. Moreover, we assessed A2B-mediated induction of MUC2 and MUC5AC expression during epithelial injury. A2B agonist treatment of wounded intestinal epithelium resulted in increased MUC2 and MUC5AC mRNA expression and MUC5AC protein expression. We further explored the A2B-mediated effects on mucus growth and penetrability during homeostasis by an ex-vivo approach.

Conclusion

Our preliminary findings suggest that signaling through the A2BAR can increase MUC2 and MUC5AC expression in the intestine during homeostasis and injury.
P0155 Investigating the effect of a novel bio-based tissue glue for Perianal Fistulas in Crohn's Disease: A Low-Cost Rodent model study
Authors: Raghunathan N , Tokala R , Nagendra B , Pillikala M , Bera A K , Adhikari J , Reddy H , B Lakshmi J , Midde S , Singh J , Chavan S , Patel R , Sekaran A , Mitnala S , Pati F , Banerjee R
Published: January 2025

Background

Perianal fistulas (PAFs) are a severe complication of Crohn’s disease (CD) significantly affecting quality of life with limited treatment options. TNBS-induced colitis rodent models to study CD-PAF is costly and time-intensive. This study aimed to study the efficacy of a novel decellularized dermal extracellular matrix (ddECM) derived from goat skin as a tissue glue 1 for PAF treatment in a low-cost acetic-acid induced CD-PAF rodent model.

Methods

Two fistula tracts (0.3mm,1.3mm from the anal sphincter) were created in 12 male Wistar rats (aged 8-10weeks, body weight 180-220g); and a seton was placed to keep them open(Fig1A) 2. 500µL of 5%(v/v)acetic acid was injected biweekly intrarectally for a week to induce colitis and fistula discharge . Ultrasound (USG), in vivo imaging (IVIS) using indocyanine green fluorescent dye, and histopathology were used to assess inflammation and fistula formation(Fig2). 1ml bio-based tissue glue was applied to occlude each of the orifices and polymerized with UV light for 30-seconds. Rats were monitored for one week post glue treatment with single doze of 5% acetic acid as maintenance. USG was repeated to confirm healing. Upon sacrifice, tissue was collected for histopathology examination to confirm the success of CD-PAF and healing.

Results

All rats exhibited soft feces, diarrhoea, widening of fistula orifice, purulent fistula discharge, blood per rectum and significant weight loss after induction(Fig1B). USG confirmed active inflammation revealing edematous fistula tracts on both sides. This was validated by strong fluorescent signal in IVIS at the site of inflammation. Histopathology staining also showed acute inflammation (Fig2). One rat died post-induction. Tissue glue was applied to the remaining 11 rats. Upon one week follow-up, 10 of 11 rats exhibited normal feces, closure of fistula, absence of purulent fistula discharge and weight gain. USG and histopathology showed complete fistula healing in 10 rats and partial healing in one.

Conclusion

This study has demonstrated the use of low-cost CD-PAF rodent model which offers a faster induction (1-week) than conventional TNBS models (8 weeks). The novel bio-based tissue glue has shown to be a feasible and an effective treatment for CD-PAFs.

References

1. Bera, A.K. et al. BioScience.2022

2. Ling Li et al. Sci. Adv. 2023

3. Singh A. et al. Inflamm Bowel Dis. 2023
P0156 Plasma immune protein profiles in therapy-naïve ulcerative colitis relate to extent of affected tissue and density of intestinal infiltration, differentiating patients with discreet disease courses
Authors: Calado B , Heredia M , Charrout M , Klomberg R C W , Barendregt D , Hulleman-van Haaften D H , Tuk B , van Berkel L A , Folly B B , Escher J C , de Ridder L , Samsom J N
Published: January 2025

Background

Ulcerative colitis (UC) is a chronic intestinal inflammation driven by inflammatory memory T helper cell (ThM) reactivation. Heterogeneity in clinical disease and variability in response to treatment require new strategies targeting the patient’s underlying immune disease. As inflammation in UC only affects the intestinal mucosa, it is debated whether peripheral blood immune responses could reliably reflect ongoing intestinal immune disease.

Methods

To unequivocally establish this, we performed a uniquely deep immune characterization of therapy-naïve pediatric UC patients (n=34) and controls (n=55) by measuring 92 plasma immune proteins, circulating ThM subpopulations, intestinal mRNA expression, and intestinal immunohistochemistry. Extensive clinical follow-up was used to relate immune responses to clinical disease severity and therapy response.

Results

At diagnosis, 23 plasma proteins were differentially abundant in UC compared to controls. Increased protein concentrations mainly related to the IL-17 pathway (IL-17, IL-8, CCL20), tissue remodelling (MMP-10, MMP1, HGF), acute phase response (IL-6, OSM) and chemokinesis (CXCL9, CXCL10, CCL3). Interestingly, intestinal mRNA expression of highly abundant plasma proteins similarly increased, whereas no proportional changes in circulating ThM were detected. Unbiased hierarchical clustering of plasma immune profiles yielded two clusters of UC patients; UC_A with significantly increased concentrations of 17 proteins, many of which Th17/neutrophil-related, versus UC_B with lower concentrations. Notable increases in UC_A were: IL- 17A; OSM, a neutrophil product previously associated with anti-TNF therapy resistance; CXCL9 and CXCL11, IFN-g-induced chemokines. Strikingly, this UC_A immune profile strongly related to increased size of the affected intestinal surface and degree of infiltrating IL-17A+ cells and number of neutrophils in the biopsies. Clinically, UC_A patients had higher clinico-pathological parameters, more severe intestinal disease at endoscopy and on histology , and required earlier treatment intensification during 1.5 year follow-up, compared to UC_B patients.

Conclusion

These data firmly establish that, even though inflammation in UC only affects the intestinal mucosa, peripheral blood immune protein responses reliably reflect the degree of ongoing intestinal immune disease in UC. Implementing immune profiling in routine clinical characterization at diagnosis improves identification of patients who will develop a severe disease course allowing tailored treatment with earlier start of more intensive immunomodulatory treatment to treat to target and avoid tissue damage.
P0157 Preoperative emotional dysfunction is associated with postoperative morbidity in Crohn’s disease patients
Authors: Schmitz A , Bercher M , Iesalnieks I
Published: January 2025

Background

Anxiety has been found to be associated with postoperative morbidity in patients with Crohn’s disease. The impact of preoperative emotional dysfunction was analyzed in the present prospective observational study.

Methods

Consecutive patients undergoing bowel resection for Crohn’s disease were asked to fill out the IBDQ-32 questionnaire. The 32-item Inflammatory Bowel Disease Questionnaire (IBDQ-32) is the most frequently used instrument to capture disease- specific quality of life. One of 4 domains scores the emotional function. The domain consists of 12 items. All items use 7-point Likert-type scales for capturing symptom- related experiences over the previous 2 weeks, with 1 indicating the highest symptom frequency/severity and 7 indicating the lowest symptom frequency/severity. For emotional function, scores between 12 and 84 can be calculated, higher score indicating better quality of life.

Results

112 patients underwent bowel resection (90 ileocolic resections, 11 colorectal resections, 5 both ileocolic and colorectal resections and 6 small bowel resections. Postoperative complications occurred in 26 patients (23%), postoperative ileus in 10 (9%), wound complications in 9 (8%) and intraabdominal septic complications in 5 (4.5%). Patients who developed postoperative complications revealed significantly decreased emotional scoring (40.3 vs. 48.3, p=0.009). Also, patients with postoperative ileus (38.2 vs. 47.1, p=0.049), wound complications (35.1 vs. 47.4, p=0.009) and intraabdominal septic complications (40.8 vs. 46.6, p=0.35) demonstrated higher grade of emotional dysfunction preoperatively. Preoperative abscess, severe preoperative anemia and preoperative weight loss were all significantly associated with an impaired preoperative emotional functioning.

Conclusion

Preoperative emotional dysfunction was associated with an increased risk of postoperative morbidity. However, severe disease complications (abscess, anemia,weight loss) led also to more emotional suffering preoperatively.
P0161 The primary sclerosing cholangitis and ulcerative colitis colonic mucosa defined through paired microbial and single-cell RNA sequencing
Authors: Tearle J , Zhang F , Jackson K , Malhotra P , Tavakoli P , Koentgen S , Warren J , Williams C , Haque A , Arzivian A , Tedla N , Kim A , King H , Hold G , Ghaly S , James K
Published: January 2025

Background

Primary sclerosing cholangitis (PSC) is a chronic progressing cholestatic disease that often co-occurs with inflammatory bowel disease (PSC-IBD). PSC-IBD affecting the colon (PSC-UC) is likened clinically to ulcerative colitis (UC), however differences include a right colon dominance, less severe inflammatory presentation and a greater lifetime risk of colorectal cancer.

Methods

We combine single-cell mRNA and antigen receptor sequencing, 16S ribosomal RNA gene analysis and spatial transcriptomics to profile mucosal biopsies from four colon regions of PSC-UC and UC patients in remission or at the time of relapse.

Results

In contrast to UC, PSC-UC patients display an enrichment of CD8 T, γδ T and natural killer cells and reduced microbial diversity in the right colon, even in the absence of symptomatic inflammation. During flare, we demonstrate the expansion of a novel inflammatory mast cell state in both diseases and highlight the function of TMEM176B in sustaining this activated state.

Conclusion

These results highlight that the PSC-UC and UC colonic mucosa are fundamentally different with shared cell programs during active disease, providing insights to guide tailored clinical management and precision therapies.
P0163 Novel IBD treatment: Small molecule oral GPRx agonists for Treg cell therapy
Authors: Zhou K , Shi Y , Wang T , Fan F , Xia T
Published: January 2025

Background

Our study has revealed that GPRx regulates gut DC migration to local draining lymph node to induce iTreg (Fig 1).1 To fulfil our drive to drug discovery and development for IBD (and other autoimmune diseases) program, we’ve rationally designed small molecule oral GPRx agonists that can induce gut antigen specific iTreg and intended to examine their efficacy utilizing TNBS induced acute IBD model.2-4

Methods

C57/BL6 8-10 weeks old female mouse was intrarectally administered with 2,4,6-trinitrobenzene sulfonic acid (TNBS) in 30% ethanol solution through a special catheter to induce colitis, from day 0 to day 4. The tested drug was given via PO, from day 0 to day 4. The experiment ended on day 5. DAI (disease activity index) on weight loss, diarrhea, and rectal bleeding, was recorded daily, to determine disease severity. Colon was taken at the end of experiment for measurement and photography.

Results

A series of compounds have been tested using this in vivo TNBS induced acute IBD mouse model and some of them have shown excellent activity (Fig 2).

Conclusion

With this new MOA and first-in-class approach using small molecule oral GPRx agonists for Treg cell therapy, we could develop a different treatment for IBD (and other autoimmune diseases in general) patients than JAK inhibition or a4b7 inhibition or S1P1 activation, etc. The outstanding efficacy data described here together with their safety profile and acute/sub-acute toxicity results have provided us the confidence to a successful IND late-2025.

References

1. Unpublished data

2. ZL2017107741357

3. PCT/CN2023/135952

4. PCT/CN2023/135912
P0166 Changes in bone mineral density in young patients with inflammatory bowel diseases
Authors: Myasoutova E , Shamsutdinova N , Abdulganieva D
Published: January 2025

Background

Inflammatory bowel diseases (IBD), which include Crohn’s disease (CD) and ulcerative colitis (UC), are an urgent problem of modern gastroenterology, which is associated with an increase in morbidity, a lifelong recurrent course, and the prevalence of the disease mainly among young people of working age. Extracellular manifestations in CD and UC include damage to the bone system in the form of impaired bone mineralization with the possible development of osteoporosis (OP). In this regard, patients with IBD have a high risk of developing osteoporotic fractures.

Methods

The aim of the study was to evaluate the absolute values of bone mineral density (BMD) in young patients with IBD.

96 patients with a verified diagnosis of IBD were examined: 41 patients had CD, 55 had UC; men -50, women - 46. The average age of the patients was 29 [25;35] years. The average age of IBD debut was 24 [19.5;29] years. All patients underwent a general clinical examination, as well as a two-adsorption X-ray densitometric examination of the lumbar spine and proximal femur on the right and left with the determination of BMD and bone mineral content (BMC). The data obtained were correlated with the NHANES III database.

Results

The average index of the Z-criterion (L1-L4) was -0.4 [-1.4;0.4]. The average Z index (hip neck) was -0.4 [-1.2;0.5]. The average BMD value at the level of the lumbar spine (L1-L4) was 1.175 [1.07;1.28] (g/cm2). The average total BMC was 53.49 [45.83;60.04] g. The average BMD value at the level of the neck of the right femur was 0.99 [0.88;1.09] (g/cm2). The average value of the total BMC in the same place was 24.86 [22.35;28.58] g. The average BMD value at the level of the neck of the left femur was 0.97 [0.88;1.09]. The average total BMC value was 25.75 [22.34;28.82]. 13.5% (13 out of 96 patients) showed a decrease in bone mass below the chronological age.

Conclusion

Despite the young age of the patient, a significant proportion of patients (13.5%) have deviations in BMD from normal values. Also, the absolute values of BMD and bone mineral content differ from the values when compared with the reference interval in the NHANES III database.

References

1.Kreienbuehl A. S. et al. Bone health in patients with inflammatory bowel disease //Swiss medical weekly. – 2024. – Т. 154. – №. 3407. DOI: 10.57187/s.3407

2.A.Kh. RAMAZANOVA, I.G. MUSTAFIN, A.Kh. ODINTSOVA, R.M. NABIULLINA, D.T. SIRAZETDINOV, D.I. ABDULGANIEVA Inflammatory bowel diseases: epidemiology and development of thromboembolic complications, Practical medicine №4 (96) 08/2016/volume 2; 96: 88-91
P0174 Enteric glial cells-derived NGF prevents necroptosis of intestinal epithelial cells and alleviates DSS-induced colitis
Authors: Min Z , Zhang Y , Hu C , Song X , Guo H
Published: January 2025

Background

The maintenance and repair of the intestinal epithelial barrier (IEB) is a multifaceted process that requires the coordinated participation of different cell types within the gastrointestinal tract. Studies have shown that ablation of enteric glial cells (EGCs) can lead to intestinal epithelial cell necrosis and destruction of the IEB. However, the specific role of EGCs in it remains ambiguous. The aim of this study was to elucidate the potential role of EGCs in regulating IEB and intestinal inflammation.

Methods

In this study，we employed immunofluorescence, western blot, ELISA, and gene expression profiling to assess the expression of GFAP (a marker for EGCs), NGF, and necroptosis-associated proteins in intestinal tissues from IBD patients. In addition, DSS-induced colitis mouse models, in vitro co-culture systems, and transcriptomic analysis were used to explore the effects of EGCs and NGF on IECs necroptosis and inflammation.

Results

In intestinal tissues of IBD patients, reduced expression of GFAP and S100β suggests damage to EGCs. Additionally, we found that NGF in intestinal tissues primarily originates from EGCs. Diminished NGF secretion due to EGCs damage correlates with increased necroptosis of IECs. Exogenous administration of NGF inhibits DSS-induced necroptosis of IECs and alleviates colonic inflammation. Ablation of EGCs exacerbates necroptosis of IECs, disruption of IEB, and intestinal inflammation following DSS treatment. In vitro co-culture experiments demonstrate that EGCs suppress necroptosis of IECs induced by T/S/Z mix. Further flow cytometry and TEM results confirm that NGF effectively mitigates T/S/Z mix-induced necroptosis of IECs.Transcriptomic sequencing indicated that RNF126 is a downstream target in the NGF signaling pathway, potentially mediating the protective effects of EGCs-derived NGF on IECs.

Conclusion

Based on the findings from this study, it is evident that EGCs play a crucial role in maintaining intestinal homeostasis through their production of NGF. Reduced NGF secretion from damaged EGCs correlates with increased necroptosis of IECs, exacerbating inflammation in colonic tissues. Exogenous NGF administration effectively attenuates this process, highlighting its therapeutic potential in IBD. Furthermore, transcriptomic analysis identified RNF126 as a downstream mediator in the NGF signaling pathway, its specific role remains speculative and requires further investigation to elucidate its mechanistic involvement in mediating the protective effects of NGF. In conclusion, these findings emphasize the intricate interplay between EGCs, NGF signaling, and IEC survival, pointing towards novel therapeutic strategies targeting this pathway in IBD management.
P0178 Lack of DNA methyltransferase DNMT3A in fibroblasts impairs cell growth and wound healing
Authors: Escudero-Hernández C , Hübner E , Núñez-Torvisco S , Rodríguez-Peña A , Montague J , Rosenstiel P
Published: January 2025

Background

Fibrosis in Crohn’s disease (CD) is a complication of chronic inflammatory processes that may result in impaired gastrointestinal function and bowel strictures. However, exact pathomechanisms leading to intestinal fibrosis in CD are not fully understood. Since the de novo DNA methyltransferase DNMT3A is involved in cellular differentiation processes and genetic variants have been associated with increased risk for CD, we sought to characterize how DNMT3A loss might affect fibroblasts, the key mesenchymal cells involved in fibrosis.

Methods

We stably deleted Dnmt3a in 3T3 fibroblast cell line using CRISPR-Cas9 technology and subsequently selected GPF-positive clones using FACS sorting. Cell cultures were performed in cell culture plates with or without 0.2% gelatin coat. Reverse transcription quantitative PCR, western blot, and immunofluorescence were used to confirm Dnmt3a gene deletion and characterize wild-type (WT) and knockout (KO) cells. Cell adhesion was evaluated by fluorescence levels of calcein acetoxymethyl ester (CAM)-labelled cells that attached to cell culture plates after 1h of culture and subsequent wash. Collagen lattice assays in presence of proinflammatory and profibrotic stimuli served to assess fibroblast contractibility.

Results

3T3 fibroblasts lacking Dnmt3a acquire an activated myofibroblast phenotype (increased a-SMA expression) that also overexpress tissue remodeling matrix metalloproteinase Mmp3, and the inflammation-induced necroptosis marker RIPK3 when compared to their WT counterparts. In addition, Dnmt3a-depleted 3T3 cells adhere more strongly to cell culture plates. In presence of transforming growth factor (TGF)-b, cells embedded in a 3D collagen matrix and lacking Dnmt3a build an interconnected cellular network and contract up to a 77.78% (compared to interconnected cells that reach up to 13.73% contraction by WT cells). This contraction was independent of the presence of interleukin (IL)-11. Profibrotic factors such as bleomycin prevent intercellular network formation in both WT and KO cells; hence, incapacitating cells to contract the collagen matrix.

Conclusion

We conclude that Dnmt3a is crucial for fibroblast homeostasis, including cell activation and functionality. Furthermore, TGF-b induces higher fibroblast contractibility when Dnmt3a is lacking. Thus, additional characterization of Dnmt3a depletion in fibroblasts will deepen the understanding of fibrosis and may point towards new therapeutic approaches for this untreatable complication.

References

1.Hampe J, Franke A, Rosenstiel P, et al. A genome-wide association scan of nonsynonymous SNPs identifies a susceptibility variant for Crohn disease in ATG16L1. Nat Genet. 2007;39(2):207-211. doi:10.1038/ng1954

2.Fazio A, Bordoni D, Kuiper JWP, et al. DNA methyltransferase 3A controls intestinal epithelial barrier function and regeneration in the colon. Nat Commun. 2022;13(1):6266. doi:10.1038/s41467-022-33844-2
P0181 Modulating macrophage activity via NAAA pharmacological inhibition: a therapeutic strategy for intestinal fibrosis
Authors: Nani M , De Cicco P , Cattaneo F , Petrosino S , Tropeano F P , Miraglia M , Pagano E , Rinaldi M M , Izzo A A , Romano B ,   Francesca Paola Tropeano
Published: January 2025

Background

Intestinal fibrosis, a frequent complication of inflammatory bowel disease, is characterized by stricture formation with no pharmacological treatment to date. N-acylethanolamine acid amidase (NAAA) is responsible for the hydrolysis of acylethanolamides (AEs, eg, palmitoylethanolamide and oleoylethanolamide). Here, we investigated NAAA and AE signalling in gut fibrosis.

Methods

NAAA and AE signalling were evaluated in human intestinal specimens from patients with stenotic Crohn’s disease (CD). Gut fibrosis was induced by 2,4,6-trinitrobenzenesulfonic acid, monitored by colonoscopy, and assessed by qRT-PCR, histological analyses, and confocal microscopy. Immune cells in mesenteric lymph nodes were analysed by FACS. Colonic fibroblasts were cultured in conditioned media derived from polarized or non-polarized bone marrow-derived macrophages (BMDMs). IL-23 signalling was evaluated by qRT-PCR, ELISA, FACS, and western blot in BMDMs and in lamina propria CX3CR1+ cells.

Results

In ileocolonic human CD strictures, increased transcript expression of NAAA was observed with a decrease in its substrates oleoylethanolamide and palmitoylethanolamide. NAAA inhibition reduced intestinal fibrosis in vivo. More in-depth studies revealed modulation of the immune response related to IL-23 following NAAA inhibition. The antifibrotic actions of NAAA inhibition are mediated by Mφ and M2 macrophages that indirectly affect fibroblast collagenogenesis. NAAA inhibitor AM9053 normalized IL-23 signalling in BMDMs and in lamina propria CX3CR1+ cells.

Conclusion

Our findings provide new insights into the pathophysiological mechanism of intestinal fibrosis and identify NAAA as a promising target for the development of therapeutic treatments to alleviate CD-related fibrosis.
P0182 The systemic administration of the anti-MCL1, S63845, reduces inflammation and fibrosis in a murine model of DSS-Chronic Colitis
Authors: Jarén A , Macías-Ceja D C , Burgos R , Cosín-Roger J , Cejudo-Garcés A , Patel J , Ortiz-Masiá D , Calatayud S , Barrachina M D
Published: January 2025

Background

A recent RNA sequencing analyses performed in ileal resections from complicated Crohn´s disease patients revealed the up-regulation of the anti-apoptotic molecule, MCL1 in fibrotic lesions. Immunohistochemical studies localized MCL1 expression in several cell types, including fibroblasts and leukocytes1. We analyse here the role played by MCL1 on inflammation and fibrosis in a murine model of chronic colitis.

Methods

C57Bl/6 mice (20–25 g) were administered intraperitoneally with Vehicle or S63845 (25 mg/kg), five days a week during the two DSS cycles (a cycle consists in 7 days drinking water with 2% DSS, followed by ten days drinking water). Body weight and Disease Activity Index (DAI) were monitored daily, and mice were sacrificed 34 days after the beginning of the experiment. A sample from the colon was paraffin embedded for both histological analysis (epithelial damage was quantified by the Obermeier score in H&E-stained slides and collagen deposition was quantified by the QuPath software in Syrius Red- stained slides) and immunohistochemistry (CD68, CD3, MPO, TUNEL and Cleaved-caspase 3). Other sample was used for RT-PCR studies. Unless otherwise mentioned, data are expressed as mean±SEM.

Results

DSS-Vh treated mice exhibited a reduction in body weight (16.9±1.0) and increase in DAI score (2.5±0.6) which reached a statistical significance (P<0.001&, P<0.01&, respectively) at day 11, compared with body weight and DAI at day 0 (19.4±0.4 and 0, respectively). In S63845-DSS treated mice, no significant changes in body weight and DAI were detected all along the experimental period when compared with the respective day 0. The administration of S63845, compared with Vh in DSS-treated mice induced: a) a reduction in epithelial damage (1.3±0.8 vs 1.7±0.4) and a significant diminution in the thickness of the submucosa (19.6±1.7 vs 42.4±3.2, respectively, P =0.0008*) and collagen deposition in the same area (median an IQ range: 54.2 (55.9-68.5) vs 61.9 (55.9-68.5) respectively, P = 0.051#). b) a lower immunostaining for both MPO and CD3; c) a similar immunostaining for CD68, TUNEL and Cleaved-caspase 3 which was very scarce for the last two and d) a lower mRNA expression of inflammatory cytokines (il1β, il6 and chi3l1), chemokines (cxcl1 and cxcl5) and profibrotic markers (col1a1, col3a1, col4a1, and tgfβ1). *t-test; #Mann-Whitnney; &Repeated measures ANOVA

Conclusion

The systemic administration of the MCL1 inhibitor, S63845 prevented the infiltration of both neutrophils and CD3 lymphocytes and failed to induce apoptosis in the affected intestine while it reduced fibrosis in a murine model of DSS-chronic colitis

References

1. Seco-Cervera M et al Biochim Biophys Acta Mol Basis Dis. 2024 Feb;1870(2):166966BBA
P0188 The Role and Mechanism of Ephrin B2 in Crohn’s Disease-Associated Intestinal Fibrosis
Authors: Hu S , Yu Q , Wang X , Xu D , Chen M , Chen Y
Published: January 2025

Background

Crohn’s disease (CD) is a chronic inflammatory bowel condition characterized by transmural inflammation that can lead to intestinal fibrosis and strictures. This study aimed to investigate the role of Ephrin B2, a member of the ephrin family, in the pathogenesis of CD-associated intestinal fibrosis.The primary objective of this study was to elucidate the expression pattern of Ephrin B2 in CD fibrosis and to explore its potential role in the modulation of autophagy and the AKT/mTOR signaling pathway, which were implicated in the fibrotic process.

Methods

The study used in vivo and in vitro methodologies to dissect the role of Ephrin B2 in intestinal fibrosis associated with Crohn’s disease. In vivo studies involved the generation of Ephrin B2 knockout mice and the induction of intestinal fibrosis using DSS-induced mice to assess the relationship between Ephrin B2 and intestinal fibrosis. In vitro studies utilized recombinant Ephrin B2 protein stimulation of CCD-18co cell lines to observe changes in autophagy-related proteins and the effects on fibroblast proliferation, activation, apoptosis, and collagen deposition. The study also investigated the impact of Ephrin B2 on signaling pathways such as PI3K/Akt/mTOR and RhoA/ROCK.

Results

Preliminary findings indicated that Ephrin B2 expression was significantly elevated in CD fibrosis patients and was associated with the modulation of intestinal autophagy genes. The in vivo model suggested a correlation between Ephrin B2 and the severity of intestinal fibrosis. In vitro studies showed that Ephrin B2 influenced the activation of fibroblasts and the deposition of collagen, potentially through the regulation of autophagy and the AKT/mTOR pathway.

Conclusion

This study provided evidence that Ephrin B2 might play a crucial role in CD-associated intestinal fibrosis. The findings suggested that targeting Ephrin B2 could be a novel therapeutic strategy for managing CD fibrosis. Further research is warranted to fully understand the mechanistic links between Ephrin B2, autophagy, and fibrosis in CD.