Active Ulcerative Colitis Patients Research Articles

Microbial metabolite deoxycholic acid-mediated ferroptosis exacerbates high-fat diet-induced colonic inflammation

High-fat diet (HFD) has long been recognized as risk factors for the development and progression of ulcerative colitis (UC), but the exact mechanism remained elusive. Here, HFD increased intestinal deoxycholic acid (DCA) levels, and DCA further exacerbated colonic inflammation. Transcriptome analysis revealed that DCA triggered ferroptosis pathway in colitis mice. Mechanistically, DCA upregulated hypoxia-inducible factor-2α (HIF-2α) and divalent metal transporter-1 (DMT1) expression, causing the ferrous ions accumulation and ferroptosis in intestinal epithelial cells, which was reversed by ferroptosis inhibitor ferrostatin-1. DCA failed to promote colitis and ferroptosis in intestine-specific HIF-2α-null mice. Notably, byak-angelicin inhibited DCA-induced pro-inflammatory and pro-ferroptotic effects through blocking the up-regulation of HIF-2α by DCA. Moreover, fat intake was positively correlated with disease activity in UC patients consuming HFD, with ferroptosis being more pronounced. Collectively, our findings demonstrated that HFD exacerbated colonic inflammation by promoting DCA-mediated ferroptosis, providing new insights into diet-related bile acid dysregulation in UC.

Exploring the autophagy-related pathogenesis of active ulcerative colitis.

The pathogenesis of ulcerative colitis (UC) is complex, and recent therapeutic advances remain unable to fully alleviate the condition. To inform the development of novel UC treatments, bioinformatics was used to explore the autophagy-related pathogenesis associated with the active phase of UC. The GEO database was searched for UC-related datasets that included healthy controls who met the screening criteria. Differential analysis was conducted to obtain differentially expressed genes (DEGs). Autophagy-related targets were collected and intersected with the DEGs to identiy differentially expressed autophagy-related genes (DEARGs) associated with active UC. DEARGs were then subjected to KEGG, GO, and DisGeNET disease enrichment analyses using R software. Differential analysis of immune infiltrating cells was performed using the CiberSort algorithm. The least absolute shrinkage and selection operator algorithm and protein-protein interaction network were used to narrow down the DEARGs, and the top five targets in the Dgree ranking were designated as core targets. A total of 4822 DEGs were obtained, of which 58 were classified as DEARGs. SERPINA1, BAG3, HSPA5, CASP1, and CX3CL1 were identified as core targets. GO enrichment analysis revealed that DEARGs were primarily enriched in processes related to autophagy regulation and macroautophagy. KEGG enrichment analysis showed that DEARGs were predominantly associated with NOD-like receptor signaling and other signaling pathways. Disease enrichment analysis indicated that DEARGs were significantly linked to diseases such as malignant glioma and middle cerebral artery occlusion. Immune infiltration analysis demonstrated a higher presence of immune cells like activated memory CD4 T cells and follicular helper T cells in active UC patients than in healthy controls. Autophagy is closely related to the active phase of UC and the potential targets obtained from the analysis in this study may provide new insight into the treatment of active UC patients.

Constituents of stable commensal microbiota imply diverse colonic epithelial cell reactivity in patients with ulcerative colitis

BackgroundDespite extensive research on microbiome alterations in ulcerative colitis (UC), the role of the constituent stable microbiota remains unclear.ResultsThis study, employing 16S rRNA-gene sequencing, uncovers a persistent microbial imbalance in both active and quiescent UC patients compared to healthy controls. Using co-occurrence and differential abundance analysis, the study highlights microbial constituents, featuring Phocaeicola, Collinsella, Roseburia, Holdemanella, and Bacteroides, that are not affected during the course of UC. Co-cultivation experiments, utilizing commensal Escherichia coli and Phocaeicola vulgatus, were conducted with intestinal epithelial organoids derived from active UC patients and controls. These experiments reveal a tendency for a differential response in tight junction formation and maintenance in colonic epithelial cells, without inducing pathogen recognition and stress responses, offering further insights into the roles of these microorganisms in UC pathogenesis. These experiments also uncover high variation in patients’ response to the same bacteria, which indicate the need for more comprehensive, stratified analyses with an expanded sample size.ConclusionThis study reveals that a substantial part of the gut microbiota remains stable throughout progression of UC. Functional experiments suggest that members of core microbiota – Escherichia coli and Phocaeicola vulgatus – potentially differentially regulate the expression of tight junction gene in the colonic epithelium of UC patients and healthy individuals.

DNA methylation fine-tunes pro-and anti-inflammatory signalling pathways in inactive ulcerative colitis tissue biopsies

DNA methylation has been implied to play a role in the immune dysfunction associated with inflammatory bowel disease (IBD) and the disease development of ulcerative colitis (UC). Changes of the DNA methylation and correlated gene expression in patient samples with inactive UC might reveal possible regulatory features important for further treatment options for UC. Targeted bisulfite sequencing and whole transcriptome sequencing were performed on mucosal biopsies from patients with active UC (UC, n = 14), inactive UC (RM, n = 20), and non-IBD patients which served as controls (NN, n = 11). The differentially methylated regions (DMRs) were identified by DMRseq. Correlation analysis was performed between DMRs and their nearest differentially expressed genes (DEGs). Principal component analysis (PCA) was performed based on correlated DMR regulated genes. DMR regulated genes then were functional annotated. Cell-type deconvolutions were performed based on methylation levels. The comparisons revealed a total of 38 methylation-regulated genes in inactive UC that are potentially regulated by DMRs (correlation p value < 0.1). Several methylation-regulated genes could be identified in inactive UC participating in IL-10 and cytokine signalling pathways such as IL1B and STAT3. DNA methylation events in inactive UC seem to be fine-tuned by the balancing pro- and anti- inflammatory pathways to maintain a prevailed healing process to restore dynamic epithelium homeostasis.

Identified S100A9 as a target for diagnosis and treatment of ulcerative colitis by bioinformatics analysis

Ulcerative colitis (UC) is a chronic, recurrent inflammatory bowel disease. UC confronts with severe challenges including the unclear pathogenesis and lack of specific diagnostic markers, demanding for identifying predictive biomarkers for UC diagnosis and treatment. We perform immune infiltration and weighted gene co-expression network analysis on gene expression profiles of active UC, inactive UC, and normal controls to identify UC related immune cell and hub genes. Neutrophils, M1 macrophages, activated dendritic cells, and activated mast cells are significantly enriched in active UC. MMP-9, CHI3L1, CXCL9, CXCL10, CXCR2 and S100A9 are identified as hub genes in active UC. Specifically, S100A9 is significantly overexpressed in mice with colitis. The receiver operating characteristic curve demonstrates the excellent performance of S100A9 expression in diagnosing active UC. Inhibition of S100A9 expression reduces DSS-induced colonic inflammation. These identified biomarkers associated with activity in UC patients enlighten the new insights of UC diagnosis and treatment.

Prolonged culturing of colonic epithelial organoids derived from healthy individuals and ulcerative colitis patients results in the decrease of LINE-1 methylation level

Patient-derived human intestinal organoids are becoming an indispensable tool for the research of digestive system in health and disease. However, very little is still known about the long-term culturing effect on global genomic methylation level in colonic epithelial organoids derived from healthy individuals as well as active and quiescent ulcerative colitis (UC) patients. In this study, we aimed to evaluate the epigenetic stability of these organoids by assessing the methylation level of LINE-1 during prolonged culturing. We found that LINE-1 region of both healthy control and UC patient colon tissues as well as corresponding epithelial organoids is highly methylated (exceeding 60%). We also showed that long-term culturing of colonic epithelial organoids generated from stem cells of healthy and diseased (both active and quiescent UC) individuals results in decrease of LINE-1 (up to 8%) methylation level, when compared to tissue of origin and short-term cultures. Moreover, we revealed that LINE-1 methylation level in sub-cultured organoids decreases at different pace depending on the patient diagnosis (healthy control, active or quiescent UC). Therefore, we propose LINE-1 as a potential and convenient biomarker for reliable assessment of global methylation status of patient-derived intestinal epithelial organoids in routine testing of ex vivo cultures.

ORGANIC ANION TRANSPORTER POLYPEPTIDE (OATP) 3A1 AMELIORATES INTESTINAL MUCOSAL DAMAGE IN INFLAMMATORY BOWEL DISEASE

Abstract BACKGROUND Inflammatory bowel disease (IBD) is a chronic condition characterised by disruption of the colonic mucosal barrier. OATP3A1/SLCO3A1 is a bile acid efflux transporter, alleviating cholestasis-associated liver injury. However, the functional role and regulational mechanism of intestinal OATP3A1 in IBD remain unclear. METHODS The expression and correlation of SLCO3A1 were investigated in the public datasets with IBD patients. Intestinal epithelial cell-specific Slco3a1 knockout (Slco3a1ΔIEC) mice were generated and fed with 4% DSS for functional studies. RESULTS Intestinal SLCO3A1 expression was significantly elevated in the ulcerative colitis (UC) and Crohn's disease (CD) patients, and positively correlated with TNFα, IL1β and MADCAM1. Compared with Slco3a1flox/flox littermates administrated by DSS, Slco3a1ΔIEC mice exhibited exacerbated colitis, characterized by more severe weight loss, enhanced colon shortening and spleen swelling. Interestingly, intestine histology assessments revealed that epithelial Oatp3a1 ablation significantly aggravated extensive disruption of the mucosal epithelium and inflammatory infiltration. The intestinal mucosal barrier function of Slco3a1ΔIEC mice was more severely damaged, as assessed by detecting serum permeated FITC-dextran. Expression of tight junction-associated proteins was decreased in Slco3a1ΔIEC mice. CONCLUSION Our data showed that intestinal OATP3A1 expression was markedly upregulated and exerted a protective role in inflammatory bowel disease. Overexpression of intestinal OATP3A1 may hold potential as a novel approach to alleviate intestinal inflammation- associated mucosal damage. Figure 1 SLCO3A1 mRNA levels in the intestinal tissues of IBD patients and generation of Slco3a1ΔIECmice. (A) SLCO3A1 mRNA expression in patients with active UC or active CD and matched controls, revealed by analyzing databases of RNA-Seq data. (B and C) Pearson’s correlation analysis between SLCO3A1 expression and TNF-α, IL1β, or MADCAM1 in colon biopsy samples from active UC (n = 74) and CD (n = 52) patients. Data were collected from GEO database GSE75214. (D) Schematic diagram of Slco3a1ΔIEC mice. (E) Genotyping of Slco3a1ΔIEC mice using genomic DNA extracted from mice tails. (F) Western blot confirmed the intestine epithelia ablation of OATP3A1 in Slco3a1ΔIEC mice. Figure 2 Effects of intestinal epithelia-specific ablation of OATP3A1 on colitis and mucosal damage in a 4% DSS treatment mouse model of IBD. (A, B) Body weight loss and spleen weight were measured from Slco3a1flox/flox or Slco3a1ΔIEC mice. (C) Serum FITC-dextran level was detected by fluorescence spectrophotometer. (D) Gross morphology images of the colon, and colon length were measured. (E) Representative images of H&amp;E-stained colons, and histopathology analysis of colitis were performed. (F) Immunoblots of colonic lysates on Zo-1, Occludin or Claudin-1(n = 4 for each group). **P &amp;lt; 0.01; ***P &amp;lt; 0.001. n = 5 for Slco3a1flox/flox group; n =6 for Slco3a1ΔIEC group treated with DSS.

P108 Response to vedolizumab in ulcerative colitis is associated with reduced neutrophil extracellular traps formation and IL-1β production: insights from NEUTROVED study

Abstract Background Vedolizumab (VDZ) is a humanized mAb against integrin α4β7. VDZ inhibits gut inflammation by impeding intestinal T-cell homing, however, recent data suggest that effects of α4β7 blocking extend beyond T-cells to alterations in innate immunity. Neutrophils are key components of innate immune response and inflammation in ulcerative colitis (UC). Studies have provided evidence linking neutrophils with UC pathogenesis, through autophagy induction and production of IL-1β-enriched neutrophil extracellular traps (NETs). NEUTROVED aimed to investigate the effects of VDZ treatment on neutrophils that may be associated with clinical and endoscopic response. Methods Patients with moderate-to-severe UC who received VDZ in standard medical care were evaluated in two timepoints, 24 hours before treatment initiation (T1) and at week 14 of treatment (T2). Clinical/endoscopic scores and common laboratory indexes were evaluated. In parallel, RNA-sequencing in peripheral neutrophils was performed, and markers of NETs in plasma and intestinal biopsies were assessed. Results In total 5 patients completed the study until today (Table 1). In T2, 4 patients demonstrated complete remission, and one patient had mild disease. No adverse events were recorded. RNA-sequencing analysis of paired samples revealed 248 significantly downregulated and 109 upregulated genes in peripheral neutrophils after VDZ treatment. Bioinformatics analysis using the GeneCodis4 web-based tool revealed that downregulated DEGs clustered mainly in immune-related pathways, including neutrophil degranulation, Toll-like receptor cascades, signaling by interleukins, NETs formation, and HIF-1 signaling pathway, while upregulated DEGs were involved in RNA metabolism-related processes, and the ribosome biogenesis pathway. A comparative analysis between these differentially expressed genes (DEGs) and our previous RNA-sequencing data of peripheral neutrophils isolated from active UC patients (n=24) highlighted a set of 168 DEGs as restored targets after VDZ therapy. Of note, several unique genes implicated in neutrophil activation, including IL-1 signaling components, were found significantly downregulated after VDZ therapy. In line with the abovementioned transcriptome alterations, markers of NETs and IL1β-bearing NETs in plasma and colonic tissue were significantly reduced, recapitalizing clinical and endoscopic response to VDZ. Conclusion Response to VDZ treatment in UC is related to anti-inflammatory actions on neutrophils characterized by reduced NETosis and IL-1β production through NETs. Further mechanistic studies investigating the possible effects of VDZ on innate immunity and neutrophil-mediated responses are warranted.

P1218 Non-invasive point-of-care stool sample for detection of Cytomegalovirus infection in patients with Active or Refractory Ulcerative Colitis using multiplex polymerase chain reaction (mPCR): a pilot feasibility study

Abstract Background Cytomegalovirus (CMV) infection is associated with more flare-ups and deteriorates the prognosis of active inflammatory bowel disease (IBD). Patients with refractory IBD should be tested for CMV colitis, especially if they are not responding to immunosuppressive therapy. The prevalence of CMV co-infection ranges from 10% to 30% in steroid-refractory acute colitis. Concurrent CMV-colitis is associated with an increased risk of worse therapeutic outcomes, resistance to corticosteroids, and need for salvage therapy. One of the two gold standards of diagnosis of CMV colitis in IBD patients with active disease requires the analysis of tissue biopsies using PCR. The aim of our pilot study was to evaluate the diagnostic accuracy of non-invasive point-of-care faecal samples using mPCR for detecting CMV in patients with ulcerative colitis compared to standard practice biopsies PCR. Methods We performed a prospective, observational, feasibility study including 62 consecutive patients with active ulcerative colitis (UC) requiring intravenous steroids or steroid-refractory UC in the Department of Gastroenterology of University Hospital „Tsaritsa Yoanna – ISUL", from April 2022 to October 2023. In all UC patients, colonic tissue biopsies were performed and analyzed by qualitative CMV PCR. Faecal sample mPCR CMV was performed on the positive patient cohort to compare the feasibility of (ME) Panel IVD using FilmArray mPCR (Biofire, bioMerieux, France). All the tested patients had undergone immunosuppressive therapy and initial stool specimen cultures were obtained to exclude infection with C. difficile and other pathogen flora. Results Out of 62 active UC patients with colonic tissue samples qualitative CMV PCR, n=5 (8%) were positive. Of these n=5 UC patients, n=4 (80%) were positive from the Fecal sample mPCR CMV, resulting in a compatibility of 80% for diagnosing CMV in these patients by the two DNA extraction methods. Calculated Sensitivity and diagnostic accuracy was 80% (95% CI) for the mPCR CMV. The patient cohort predominantly comprised males (75%) with a median age of 45 years (range: 30–72 years). Three patients (75%) out of this cohort group were treated with peroral antiviral agent Valganciclovir, one (25%) of them went into clinical and endoscopic remission after the treatment and two patients (50%) received colectomy. The median time performing the Faecal sample mPCR CMV was approx. 1 hour. Conclusion This novel faecal mPCR point-of-care test is a promising, fast and easy non-invasive tool for detection CMV in active or refractory UC patients, comparable with the colonic tissue CMV-DNA PCR.

P204 Utility of Rotational Thrombo-Elastometry as an Objective Measure of Hypercoagulability in Patients with Ulcerative Colitis-A Prospective Case Control Study

Abstract Background Inflammatory bowel disease (IBD) has an increased risk of venous thromboembolism (VTE) with factors like hospitalization and surgery increasing this risk. Although few recent guidelines recommend anticoagulation thromboprophylaxis for hospitalised IBD patients, there is wide variation in reported practices for VTE prophylaxis. This study was aimed to evaluate Rotational Thrombo-Elastometry (ROTEM) for assessing blood coagulation status in ulcerative colitis (UC) and to evaluate the relationship between ROTEM, disease severity and response to treatment. Methods This was a prospective age- and sex-matched study with 60 patients each in UC and irritable bowel syndrome (IBS) groups, latter being controls. Clinical details, blood investigations including ROTEM and D-dimer were collected and analysed after taking informed consent. Hypercoagulable state was defined using Kaufmann’s classification. ROTEM parameters [clotting time (CT), clot formation time (CFT), alpha angle (AA), maximum clot firmness (MCF), maximum lysis (ML)] and D-dimer levels were compared between the two groups. Results Of the 60 UC patients (age: 38.6±11.64 years, 44 males); 15(25%) were in remission, 18(30%), 17(28.3%) and 10(16.7%) had mild, moderate, and severe disease respectively. The UC group had significantly more hypercoagulable ROTEM than the control group (66.7% vs. 36.7%, p=0.001). Hypercoagulable ROTEM was found in 9(60%), 8(44.4%), 14(82.3%) and 9(90%) of those in remission, with mild, moderate and severe UC respectively. Compared to controls; CT [292.36±70.45 vs 337.56±77.89, p=0.001], CFT [103.15±30.31 vs 128.61±40.18, p=0.0001], AA [69.88±5.19 vs 65.36±6.4, p=0.00004], MCF [65.75±5.64 vs 63.25±4.43, p=0.008] and D-dimer levels [329(63-2358) vs 145.5(54-1304), p=0.00001) were significantly abnormal in UC group. Among UC patients with mild, moderate and severe disease; only CFT, AA and D-dimer were significantly different between the groups. There was no significant difference in ROTEM values and D-dimer level in patients with severe UC after one week of treatment. On univariate analysis; partial Mayo score [OR: 1.49, 95% CI: 1.04-2.12, p=0.028], serum albumin [OR: 0.32, 95% CI: 0.10-0.98, p=0.04], hemoglobin [OR: 0.59, 95% CI: 0.40-0.86, p=0.007], and platelets [OR: 1, 95% CI: 1.01-1.02, p=0.035], were predictors of hypercoagulable state in active UC. On multivariate analysis; only haemoglobin [OR: 0.61, 95% CI: 0.38-0.98, p=0.04] was found to be significant independent predictor. Conclusion UC patients had hypercoagulable ROTEM regardless of activity of inflammatory process. The degree of hypercoagulability increased with disease activity. Low haemoglobin was found to predictive of hypercoagulable state in active UC patients.

P1226 Determination of oral and gut mucosal microbiome profiles of patients with treatment-naïve ulcerative colitis

Abstract Background The oral microbiome in ulcerative colitis (UC) patients and its role in the pathogenesis of the disease are still poorly understood. Although there are studies investigating the composition of oral microbiome in UC, there are no studies on concomitant analyses of oral and colonic mucosal samples in treatment-naïve patients. In this study, the oral and mucosal gut microbiome of UC patients and oral microbiota of healthy individuals (HC) were compared. Methods Sixty patients with newly-diagnosed active (mild-moderate severity) UC patients (n=30) and HC (n=30) were included in the study. Dietary habits were stable in the last three months of participants (checked by dietary recall questionnaires) and co-morbidities affecting microbiota profiles were excluded. Multiple colonic mucosa biopsies were obtained from inflamed areas before treatment in UC group. Simultaneously, subgingival plaque samples were analyzed. All samples were identified through next-generation DNA sequencing analysis, evaluated using bioinformatic tests. Results The potential signature bacterial species associated with UC were determined by examining of both gut and oral microbiomes. According to the Microbiome Lefse analysis, Prevotella copri emerged as the prominent common species in the colonic mucosal and subgingival plaque biopsies of UC group. Oral microbiome comparison between UC and HC patients revealed increased Haemophilus parainfluenza and Corynebacterium durum species and decreased F. prastnutzii and Akkermansia muciniphilia in the UC group (p&amp;lt;0.022). Comparative Boxplot analysis of bacterial abundance and alpha-diversity, as indicated by the Shannon index data, revealed that the HC group exhibited higher bacterial abundance and diversity in subgingival plaque samples than the UC group. We found significant association of Bacteroides vulgatus, Prevotella copri, Bacteroides fragilis, Parabacteroides merdae in both colonic mucosal and subgingival plaque samples (oral microbiome) of UC patients (p&amp;lt;0.044). Conclusion Our study is the first to show the oral and colonic mucosal microbiome relationship in UC. The presence of oral dysbiosis associated with UC in our study supports the hypothesis that UC could be associated not only with gut dysbiosis, as previously observed, but also with an imbalance in the oral microbial communities. Interestingly, we could not detect previously reported species of Streptococcus, Oribacterium, Rothia, Neisseria and Porphyromonas in the oral microbiome samples of the UC group. However, oral and gut microbiome profiles share some common microorganisms such as Prevotella copri and Bacteroides fragilis. Further studies are required for determining the potential role of oral dysbiosis is disease severity.

DOP45 Blood-derived neutrophils give rise to differentially activated populations in the mucosa of active Inflammatory Bowel Disease patients

Abstract Background Infiltration of neutrophils is the main histological feature in active inflammatory bowel disease (IBD). Recent data suggests that tissue- and tumour-infiltrating neutrophils can adopt remarkably different phenotypes with potentially different functions. However, the heterogeneity of this population in IBD remains unknown. Methods Single-cell RNA sequencing (scRNA-seq) data was generated from colonic biopsies of 92 individuals including 6 healthy controls (HC), 36 inactive IBD, 19 active Crohn’s disease (CD) and 31 active ulcerative colitis (UC) patients. Neutrophils (6038 cells) found in the inflamed colon of CD and UC patients were analysed using a curated pipeline in Seurat (v4.0.2)1. Posterior analysis included the FindAllMarkers function (differentially expressed genes), Jaccard index similarity and the AddModuleScore function (comparisons with literature-described neutrophils), Monocle3 algorithm (developmental trajectory) and clusterProfiler (v3.0.4; enrichment and pathway analysis). For in vitro stimulation, neutrophils were isolated from the blood of HC donors (n=4) using a double Ficoll gradient 1. Cells and supernatants were analysed for qPCR and ELISA determinations. Results We identified 6 different clusters/states of colonic neutrophils (Figure 1A). In agreement with our previous results1 we found a population of S100A12/S100A8/S100A9 neutrophils (N0), IFN-response neutrophils (N3) and CCL3/CCL4 neutrophils (N2), in addition to N1.1 and N1.2 neutrophils. N3 neutrophils were enriched in the inflamed colon of CD patients compared to UC (17.2% vs 5.8%, p-value= 0.0021; Figure 1C). Comparative analysis with published datasets revealed that N0, N1.2 and N3 resemble populations of neutrophils found in blood2,3. In contrast, N1.1 and N2-like clusters showed some similarity to umbilical cord blood3 and tumour-infiltrating neutrophils4. Trajectory analysis, originating from N0 (peripheral-like) neutrophils, showed two potential branches of differentiation, with N1.2 leading towards N3 (IFN-response) and N1.1 differentiating into N2 (CCL3) neutrophils (Figure 1B). Finally, pathway analysis showed that N2 neutrophils presented enriched gene expression of the TNF signalling pathway, and together with the N1.1 state, an enrichment of the IL1β signalling pathway as well. In vitro experiments revealed that TNF, IL1β and LPS can all induce expression of CCL3 and CCL4, top differentially expressed genes of N2 neutrophils. Conclusion We show that active IBD is associated with a highly heterogeneous population of infiltrating neutrophils. Some of them resemble neutrophils in peripheral blood, while others most likely differentiate in response to inflammatory cues present in the colon of active IBD patients.

P066 Serum amyloid A – A Novel Biomarker of Ulcerative Colitis Disease Activity

Abstract Background Despite recent advancements in the treatment of ulcerative colitis (UC), a substantial number of patients fail to achieve long-term remission. Persistent histological activity has been linked with poorer treatment outcomes. Histological remission is now an accepted treatment target; however there remains significant variability in the interpretation of UC histology. As such, there is a need for novel biomarker identification to aid assessment and ultimately predict disease relapse. Serum amyloid A (SAA) is an acute-phase protein, of which serum levels have shown promise as a biomarker in IBD1. This study aims to explore the utility of SAA levels in UC colonic tissue as a diagnostic biomarker for disease activity and progression. Methods Two cohorts were prospectively recruited, including healthy controls and UC patients. Sigmoid biopsies were collected and tissue explants generated. Tissue-conditioned media from these explants was collected and secreted SAA quantified using 54 V-plex ELISA. Demographic information, disease characteristics, endoscopic Mayo scores and disease progression were documented. Endoscopic remission was defined as a Mayo endoscopic sub-score ≤1. Disease progression was defined as the requirement for corticosteroid therapy, UC-related hospitalisation, UC-related surgery or the introduction of a new immunomodulatory agent in follow-up period. P values &amp;lt;0.05 were considered significant in analyses. Results The two cohorts included 11 healthy controls and 16 UC patients (endoscopic remission n=6). Active UC patients demonstrated significantly higher SAA concentrations than healthy controls (p=0.0013) and those in endoscopic remission (p=0.02). There was no significant difference in SAA concentrations between healthy controls and UC patients in remission (p=NS). UC patients in the lowest SAA concentration quartile had a significantly longer time to disease progression (p=0.0462) (Figure 1). Conclusion Quantification of SAA secretion in IBD ex-plants has potential as a biomarker of UC activity and progression. Further investigation of SAA as a biomarker in IBD is warranted.

DOP52 Guselkumab improves symptoms of fatigue in patients with moderately to severely active Ulcerative Colitis: Phase 3 QUASAR induction study results at Week 12

Abstract Background Fatigue is a common symptom in patients with ulcerative colitis (UC) that impairs health-related quality of life. The Phase 3 QUASAR Induction Study (NCT04033445) was a randomized, double-blind, placebo-controlled, parallel-group, multicenter study of guselkumab (GUS), an interleukin-23 p19 subunit antagonist, in patients with moderately to severely active UC. The impact of GUS IV induction on patient-reported symptoms of fatigue is reported here. Methods Patients were randomized 3:2 to receive GUS 200 mg IV or placebo IV at Weeks 0, 4, and 8. At baseline and Week 12, patients completed the Patient-Reported Outcomes Measurement Information System (PROMIS)-Fatigue Short Form 7a (SF7a) which includes 7 items to evaluate symptoms of fatigue (ie, tiredness, exhaustion, mental tiredness, and lack of energy) and associated impacts on daily activities (ie, activity limitations related to work, self-care, and exercise). PROMIS-Fatigue-SF7a raw scores were converted to a standard T-score based on a general US population of mean=50 and SD=10. Higher scores indicate more severe fatigue symptoms. PROMIS-Fatigue SF7a outcomes were evaluated at Week 12 based on the improvements of ≥3, ≥5 and ≥7 points from baseline in PROMIS-Fatigue SF7a T-score. A ≥7-point improvement was defined as fatigue response (multiplicity-controlled secondary endpoint). A cumulative distribution function (CDF) curve and a probability density function (PDF) plot were used to demonstrate consistency of fatigue response. Results Seven hundred one patients were randomized and treated in the primary analysis population (mean UC duration, 7.5yrs, Mayo endoscopy subscore=3 [severe disease], 67.9%, and mean modified Mayo score, 6.9, at baseline). The mean (SD) PROMIS-Fatigue SF7a T-scores at baseline were similar between the treatment groups: 56.0 (8.77) for GUS 200 mg IV and 56.4 (8.90) for placebo IV. At Week 12, the mean change from baseline in PROMIS-Fatigue SF7a T-score was -5.7 for GUS and -2.1 for placebo (nominal p&amp;lt;0.001). Compared with placebo at Week 12, a higher proportion of GUS-treated patients achieved fatigue response (Table 1) and clinically meaningful ≥3-point and ≥5-point improvements in PROMIS-Fatigue SF7a T-scores (56.8% and 49.4% vs 34.6% and 26.4%, respectively, both nominal p&amp;lt;0.001). Both the CDF curve and PDF plot demonstrated consistency of greater treatment effect of GUS on fatigue vs placebo (Figure 1). Conclusion Moderately to severely active UC patients who received GUS induction treatment showed greater improvement in patient-reported symptoms of fatigue at Week 12 compared with placebo-treated patients.

P1237 Development of an algorithm to identify the best donor-recipient match for FMT in IBD patients based on immune system/microbiota interactions

Abstract Background IBD are characterized by uncontrolled immune responses, in genetically predisposed individuals, to the dysbiotic microbiota. Its modulation has thus emerged as a potential therapeutic strategy . Fecal microbiota transplantation (FMT) showed encouraging results for the treatment of mild-to-moderate ulcerative colitis (UC) patients. However, currently the success rate of FMT in UC patients is around 30%, and there are no criteria to predict it. We hypothesized that the immune system-microbiota interaction can dictate which graft will be accepted or rejected. Thus, we generated an algorithm to identify the best donor-recipient match for FMT treatment in IBD patients based on the functional interaction of recipient's immune system with possible donors' microbiota . Methods Mucosal and circulating immune cells isolated from 16 active UC patients, potentially eligible for FMT, were exposed to the microbiota of a panel of 8 potential healthy FM donors collected in the FM biobank of the Policlinico Hospital, Milan. The functional profile of recipients' immune cells before and after the in vitro exposure to the possible donors’ microbiota was assessed by cytokine production, gene expression profiling and activatory or inhibitory surface molecules modulation on T cells. Sequencing of the microbiota of each individual FM donor was performed by 16S. These functional data were used to develop an algorithm to identify the best donor-recipient match based on the capability of individual donors’ microbial ecologies to simultaneously reduce the inflammatory functional profile and increase the tolerogenic functional profile of the recipient's immune cells Results We demonstrated that mucosal immune cells from UC recipients exhibited a variety of responses upon exposure to the microbiota from distinct healthy donors, which clusters in two different enterotypes. Each UC recipient exhibited specific preferences for one or more FM donor in the reduction of the inflammatory profile of immune cells and concomitantly enhancement of anti-inflammatory cytokines. On te basis of these data we generated an algorithm capable of predicting the most successful functional interaction between donors and recipients. Conclusion Our study suggest that specific functional characteristics of both the donor microbiota and the recipient's immune system should be considered when selecting an optimal donor-recipient match for FMT. This study contributes to the development of a screening tool for donor selection in FMT, enabling personalized and effective therapeutic interventions for UC patients

P537 Analysis of immunological VAriables in ex vivo Tofacitinib-treated human biopsies from Active ulcerative colitis patients to predict clinical Response (the AVATAR study)

Abstract Background Ulcerative colitis (UC) is characterized by chronically relapsing inflammation of the colonic mucosa. Tofacitinib (TOFA) is a paninhibitor of the JAK kinases, a family of intracellular kinases in volved in the signal transduction pathway of multiple inflammatory signals. Despite the efficacy shown by TOFA in inducing and maintaining remission in moderately-to-severely active UC, rates of primary non response remains high. We propose a new method to identify predictors of responsiveness to TOFA based on "ex-vivo" challenge of organ cultures from biopsies collected from candidate patients. Methods Baseline biopsies from inflamed areas of UC patients with active disease and candidate to receive TOFA 10mg b.i.d. for 8 weeks, were used for "ex-vivo" organ culture and stimulated with TOFA 20μM and 200nM or left untreated. After 24 hours, proteomic analysis was performed and quantitative expression of 92 inflammation-related human proteins (Olink® Human Inflammation panel) was correlated with week 8 clinical response defined by Modified Mayo Score (MMS). Intestinal biopsies were also collected at baseline and at 8 weeks for proteomic analysis. To decipher the nature of TOFA's effects, both unsupervised and supervised machine learning (ML) methods were employed (sklearn module, Python). The K-means method was used for unsupervised learning followed by a supervised ML approach to classify week 8 responders and non-responders. Data were transformed in into a second-order polynomial format and logistic regression was used for modelling. Results Between Jannuary 2022 and January 2023, 17 UC patients were enrolled (Table 1). At week 8, 13/17 (76.5%) patients treated with TOFA showed clinical response. Unsupervised ML, utilizing proteomic data from organ cultures stimulated with TOFA 200nM, achieved a 90% success rate in predicting a positive clinical response. This prediction was based on forming two distinct clusters, with significant contributions from key variables such as CASP-8, CCL19, CCL20, CD5, CXCL9, EN-RAGE, GDNF, HGF, IL-18R1, MCP-1, MCP-2, MCP-4, MMP-10, OPG, PD-L1, STAMBP, TNFRSF9, TRANCE, TWEAK, and VEGFA. In our supervised ML approach, logistic regression was employed with 30 shuffle splits to refine the predictive model. We discovered that transforming the dataset into a second-order polynomial format significantly enhanced the model's performance. The most effective logistic regression model achieved a mean precision score of 0.82 (Fig.1). Conclusion Our predictive model demonstrates a high level of precision in identifying positive treatment responses, emphasizing the potential of further dataset expansion and the use of advanced machine-learning to predict response in TOFA-treated UC patients.

Predictive Value of MHR and NLR for Ulcerative Colitis Disease Activity.

Numerous non-invasive serologic tests are available to diagnose and monitor ulcerative colitis (UC), but their accuracy levels are limited. Thus, there is a pressing need for a serologic biomarker with higher precision for clinical practice. This study aims to evaluate the predictive capacity of monocyte/HDL ratio (MHR) and neutrophil/lymphocyte ratio (NLR) for UC disease activity. We conducted a retrospective analysis of 81 UC patients and 77 age- and sex-matched healthy controls. UC patients were categorized into active and inactive groups based on the Mayo score. The Mayo endoscopic subscore classified them into mild-to-moderate and severe UC groups. The optimal cut-off values for diagnosing UC were 0.34 for MHR (85.7% sensitivity, 76.0% specificity, 88.9% positive predictive value, 70.4% negative predictive value) and 2.49 for NLR (66.1% sensitivity, 88.0% specificity, 92.5% positive predictive value, 53.7% negative predictive value). The optimal MHR and NLR cut-off values to differentiate between mild-to-moderate UC and severe UC were 0.38 (92.9% sensitivity, 56.6% specificity, 53.1% positive predictive value, 93.7% negative predictive value) and 3.46 (71.4% sensitivity, 88.7% specificity, 76.9% positive predictive value, 85.5% negative predictive value), respectively. NLR and MHR are simple yet effective biological predictors of disease activity in UC patients.

Identification of 6 cuproptosis-related genes for active ulcerative colitis with both diagnostic and therapeutic values.

Cuproptosis has been reported to affect a variety of diseases. Therefore, we aimed to examine the role of cuproptosis-related genes in active ulcerative colitis (UC). We acquired 2 datasets of active UC from the Gene Expression Omnibus database and created immune cell infiltrations to research immune cell dysregulation. Based on the cuproptosis gene set and differentially expressed genes (DEGs), we identified the differentially expressed genes of cuproptosis (CuDEGs). We then used 2 machine learning methods to screen hub CuDEGs. Subsequently, we performed validation on additional datasets and investigated the relationship between hub CuDEGs and drug treatments. Thirty-five controls with inactive UC and 90 patients with active UC were obtained from the training sets. A total of 9157 DEGs and 27 CuDEGs were identified, respectively. Immune cell infiltration analysis revealed that patients with active UC exhibited higher levels of activated dendritic cells and neutrophils as well as lower levels of CD8+ T cells, regulatory T cells (Tregs), and macrophage M2. A six-gene cuproptosis signature was identified using machine learning algorithms. We further validated that the 6 hub CuDEGs showed a strong correlation with active UC and acted as cuproptosis-related biomarkers of active UC. Moreover, the expression of ATOX1 was downregulated, and SUMF1, MT1G, ATP7B, FDX1, and LIAS expression was upregulated in the colonic mucosa of active UC patients who responded to golimumab or vedolizumab therapy. With the exception of ATP7B, the expression patterns of hub CuDEGs before and after infliximab treatment of patients with active UC were similar to those of golimumab and vedolizumab. Cuproptosis and active UC have a complex relationship, as illustrated in our study. ATOX1, SUMF1, MT1G, ATP7B, FDX1, and LIAS are cuproptosis-related hub genes of active UC. Our study opens new avenues for investigating UC progression and developing novel therapeutic potential targets for the disease.

S1063 Improvement in Inflammatory Biomarker Levels Through Week 12 in Moderately to Severely Active Ulcerative Colitis Patients Treated With Guselkumab: Results From the Phase 3 QUASAR Induction Study

S1063 Improvement in Inflammatory Biomarker Levels Through Week 12 in Moderately to Severely Active Ulcerative Colitis Patients Treated With Guselkumab: Results From the Phase 3 QUASAR Induction Study

Serum IL-22 as a Novel Non-Invasive Biomarker in Diagnosis and Assessment of Activity in Ulcerative Colitis Patients

Abstract Background Ulcerative colitis (UC) is a multifactorial disease of unknown etiology. Mucosal healing is an important predictor of the course of disease and has therefore become the aim of UC treatment. Repeated endoscopies to identify mucosal inflammation are expensive. Interleukin (IL)22 is a member of the IL-10 family of cytokines that was recently discovered to be mainly produced by both adaptive and innate immune cells. Aim of the Work The aim of this study was to evaluate the role of serum IL-22 as a novel noninvasive biomarker in diagnosis and assessment of activity in ulcerative colitis patients. Patients and Methods This was a case control study that was conducted on 55 subjects at Ain shams university hospitals inpatients and outpatients settings throughout 6 months. Subjects were divided into 2 groups: Group (A): that included 40 patients with ulcerative colitis: 20 patients in clinical, laboratory and endoscopic remission and 20 patients in clinical, laboratory and endoscopic activity and Group (B) that included15 healthy controls. Results Statistical analysis revealed the following: Age ranges from 32_77 years and the mean age was 36 years in this study. Males were affected slightly more than females in our study. NO statistically significant difference found between three groups regarding HB, Total bilirubin, Creatinine and Urea, and there was statistically significant difference found between three groups regarding Platelets and AST, and there was highly statistically significant difference found between three groups regarding Wbcs, ALT, ESR, CRP and Albumin. For Interleukin 22, there was highly statistically significant difference found between three groups. There was highly statistically significant difference found between IL-22 (Pg/ml) and Colonoscopy, and there was statistically significant difference found between IL-22 (Pg/ml) and Histological examination, and there was no statistically significant difference found between IL-22 (Pg/ml) and Extend. Receiver operating characteristic curve (ROC) showed that the best cut off point of IL-22 (Pg/ml) to detect Patient group was found &amp;gt; 4.7 with sensitivity of 90.0%, specificity of 100%, PPV of 100.0%, NPV of 78.9% and total accuracy of 0.95%. Conclusion It is concluded that IL22 is an important marker of ulcerative colitis as it shows significant correlation with endoscopic and histological features, so it can be used instead of these interventions to achieve high satisfaction to patients.