bluuebunny/crossref_metadata_2025 huggingface dataset:
DOP10 Intestinal Ultrasound at IBD diagnosis predicts major disease events – A Copenhagen IBD cohort study
Authors: Madsen G R , Attauabi M , Wilkens R , Ilvemark J F K F , Theede K , Bjerrum J T , Dorn-Rasmussen M , Jansson S , Wewer A V , Bendtsen F , Seidelin J B , Boysen T , Burisch J
Published: January 2022

Background

The disease course of inflammatory bowel disease (IBD) is heterogeneous and highly unpredictable. Intestinal Ultrasound (IUS) is a non-invasive modality capable of assessing disease activity in IBD. IUS is reliable, patient-friendly, and allows frequent monitoring of disease activity. However, the evidence for IUS as a predictor of disease course is still limited. Here we present novel data on the predictive value of IUS performed at the time of IBD diagnosis.

Methods

Patients with new-onset IBD are currently being included in the ongoing multicentre prospective inception cohort study, the IBD Prognosis Study. During the first five years following diagnosis, patients undergo regular clinical, biochemical, endoscopic, and imaging assessments. IBD treatment and disease events are prospectively recorded. IUS is performed at diagnosis, after three months, and hereafter annually. Patients with proctitis do not undergo assessment with IUS. The newly developed IUS score, the International Bowel Ultrasound Segmental Activity Score (IBUS-SAS), is calculated for the most inflamed segment, with a high score indicating severe disease activity. The score incorporates bowel wall thickness (BWT), bowel wall stratification, colour Doppler signal, and inflammatory fat. In this abstract, we report our preliminary results after including patients for six months.

Results

IBUS-SAS at diagnosis was available in 60 patients. 32 patients were diagnosed with Crohn’s disease (ilieal: 7, colonic: 19, ileocolonic: 6) and 28 patients were diagnosed with ulcerative colitis (UC) or unclassified IBD (proctitis: 1, left-sided colitis: 8, extensive colitis: 19). The mean IBUS-SAS at diagnosis was 51.1, with a mean BWT of 5.2 mm. Major clinical outcomes were initiation of biologic therapy, n=12 (20.0%), IBD-related bowel resection, n=5 (8.3%), IBD-related hospitalisation, n=19 (31.7%). The mean IBUS-SAS at diagnosis was 66.2 among patients with the combined endpoint of any of these disease outcomes during follow-up vs. 34.7 for no major outcomes (p<0.001), see Figure 1. Additionally, we found that all patients with an IBUS-SAS above 80 at diagnosis had been hospitalised and started on systemic steroids. So far, 20 patients had an IUS follow-up scan after three months showing a mean IBUS-SAS reduction by 17.0 points (p=0.008).

Conclusion

We present data on the predictive value of early IUS in new-onset IBD. IUS activity at diagnosis of IBD seems to have the capability to predict short term disease outcome. At diagnosis, high IBUS-SAS is associated with major disease events such as starting biological therapy, IBD-related bowel resection, and IBD-related hospitalisation. Furthermore, response to treatment is reflected by a decrease in IBUS-SAS after three months.
DOP12 Non-invasive assessment of intestinal inflammatory activity in Ulcerative Colitis by Multispectral Optoacoustic Tomography (MSOT)
Authors: Klett D , Jesper D , Vitali F , Federle A , Atreya R , Strobel D , Leppkes M , Neufert C , Rath T , Neurath M F , Waldner M J
Published: January 2022

Background

In order to guide therapy in Ulcerative Colitis (UC), repeated determination of intestinal inflammatory activity is essential. Endoscopy is the standard procedure to assess inflammation in UC. However innovative methods for non-invasive, uncomplicated and risk free estimation of inflammatory activity are needed as bowel preparation, patients discomfort and risk of procedural complications limit the (frequent) use of colonoscopy. Multispectral optoacoustic tomography (MSOT) is a promising new method to measure inflammation in UC. Using short and harmless impulses of NIR-lasers, it allows for determination of a specific hemoglobin-signal in the bowel-wall and therefore inflammatory activity in affected bowel segments. However, its informative value in UC has not been evaluated so far.

Methods

In 34 patients with confirmed UC, clinical activity parameters (e.g. clinical Mayo-Subscore, B-mode-sonography, C-reactive protein, white blood count) were collected and MSOT of the sigmoid was performed within 2 weeks before/after endoscopy. For MSOT, a commercially available clinical MSOT-system (Acuity Echo, iThera Medical, Munich) was used with sequential analysis of collected data on an external desktop PC. Finally, clinical data, ultrasound findings (Limberg) and MSOT-parameters (single wavelenghts 760 nm, 800 nm, 900 nm; multispectral signals hb, hbO2, hbT) were correlated with endoscopic findings (Mayo endoscopic Subscore, MES).

Results

We found strong and significant correlation between MES and MSOT parameters 800 nm (Spearman r = 0,6599; p < 0,0001) and HbO2 (Spearman r = 0,6695; p < 0,0001), superior to sonographic evaluation of the inflammatory activity in affected bowel segments (Spearman r = 0,4914; p = 0,0023). Simultaneously these MSOT parameters demonstrated excellent sensitivity and specifity in distinguishing moderately to highly active (MES 2,3) from inactive and mild disease (MES 0,1) (800nm: AUROC 0,9063 (p < 0,0001); sensitivity = 93,75 %, specificity = 88,89 %; HbO2: AUROC 0,9063 (p < 0,0001); sensitivity = 100 %, specificity = 88,89 %).

Conclusion

MSOT is a promising approach to non-invasively assess intestinal inflammation in UC and therefore monitor anti-inflammatory therapy in these patients. Further studies are required to validate these findings.
DOP16 Categorizing endoscopic severity of Crohn’s Disease using the Modified Multiplier SES-CD (MM-SES-CD)
Authors: Narula N , Pray C , Wong E , Colombel J F , Marshall J , Daperno M , Reinisch W , Dulai P
Published: January 2022

Background

Current endoscopic scoring indices such as the Simple Endoscopic Score for Crohn’s Disease (SES-CD) quantify the degree of mucosal inflammation in Crohn’s disease (CD) but lack prognostic potential. The Modified Multiplier of the Simple Endoscopic Score for Crohn’s Disease (MM-SES-CD) is an internally validated endoscopic scoring tool that quantifies the endoscopic burden of CD and can be accessed online (https://www.mcmasteribd.com/mm-ses-cd).1 This analysis aims to establish thresholds of the MM-SES-CD that classify CD endoscopic burden into inactive or very mild disease, mild, moderate, and severe disease based on the probability of achieving endoscopic remission (ER) on active therapy at one-year.

Methods

This post-hoc analysis included pooled data from three CD clinical trials (n=350 patients, baseline SES-CD ≥3 with confirmed ulceration). Maximum Youden Index calculations were used to determine thresholds for severity. Chi-square tests of trend were used to compare achievement of ER between severity categories, and Kaplan-Meier survival curve analysis was used to compare time to clinical remission (CR).

Results

Table 1 demonstrates the baseline characteristics of the 350 participants included in this analysis. MM-SES-CD severity categories were established as inactive or very mild (score <14), mild (score ≥14 to <31), moderate (≥31 to <45), and severe (score ≥45), which were predictive of one-year ER (50%, 30.3%, 21.7%, 8.8% respectively p<0.001) (Table 2). Lower MM-SES-CD scores had numerically higher rates of one-year CR, and time to CR over 52 weeks was superior to those with higher scores (p=0.0492) (Figure 1). MM-SES-CD thresholds for the achievement of ileal ER at one-year among 75 patients with isolated ileal disease were also established as mild (score <14), moderate (score ≥14 to <33), and severe (score ≥33), which were predictive of one-year ER (66.7%, 33.3%, 13.3%, respectively p=0.027) (Table 3).

Conclusion

We have established numerical cut-offs of the MM-SES-CD that categorize endoscopic disease severity and are prognostic for one-year ER and CR. These cut-offs could help ensure adequate balance between study arms (e.g. in prevalence of mild or moderate endoscopic disease) and identify patients with severe endoscopic disease and low probability of achieving ER.

Reference

1. Predicting endoscopic remission in Crohn’s disease by the modified multiplier SES-CD (MM-SES-CD). Gut, 2021: p. gutjnl-2020–323799.
DOP35 Increasing incidence of pouchitis among patients undergoing ileal pouch-anal anastomosis between 1996 and 2018: A population-based Danish cohort study
Authors: Barnes E , Allin K , Iversen A , Herfarth H , Jess T
Published: January 2022

Background

Current knowledge regarding the epidemiology of pouchitis is based on highly selected, mostly single-center, patient cohorts. Our objective was to prospectively determine the population-based incidence of pouchitis in patients with ulcerative colitis (UC) in the first 2 years after ileal pouch-anal anastomosis (IPAA) and analyze time trends of the incidence of pouchitis.

Methods

We used national registries to establish a population-based cohort of all Danish patients undergoing proctocolectomy with IPAA UC between 1996 and 2018. The primary outcome was the development of pouchitis within the first 2 years after IPAA, evaluated by time period of IPAA. The period of IPAA was categorized as follows: 1996–2000 (pre-biologic-1), 2001–2005 (pre-biologic-2), 2006–2010 (early-anti-TNF), 2011–2014 (expanding anti-TNF), 2015–2018 (current biologic). Pouchitis was defined using a previously developed case-finding definition for use in administrative claims data. Kaplan Meier and Cox Proportional Hazard modeling were utilized in the evaluation of time to development of pouchitis.

Results

We identified 1,664 patients that underwent an IPAA for UC. The cumulative incidence of pouchitis in the 2 years after IPAA increased throughout the study period, from 40% in 1996–2000, (95% CI:35%-46%) to 55% in 2015–2018, (95% CI:48%-63%, Table 1). Patients undergoing surgery between 2015–2018 also demonstrated an increased risk of pouchitis compared to the earliest study period (1996–2000) after adjusting for sex, age, and socioeconomic status (Hazard Ratio [HR] 1.57, 95% CI:1.20–2.05, Table 2, Figure 1). In a separate model adjusting for the same clinical and demographic factors, there was no significant relationship between the preoperative use of anti-tumor necrosis factor alpha therapy use and the risk of pouchitis when evaluated in the entire population (HR 1.14, 95% CI:0.93–1.40) and when evaluated by time period of surgery.

Conclusion

This first population-based study demonstrated a 15% absolute and 38% relative increase in the incidence of pouchitis among patients undergoing surgery between 1996 and 2018, with the greatest cumulative incidence of pouchitis demonstrated in the most recent era (2015–2018). The striking increase in the incidence of pouchitis highlights the need for further research into causes and prevention of pouchitis.
DOP38 Upadacitinib Therapy Reduces Ulcerative Colitis Symptoms as Early as Day 1
Authors: Vermeire S , Colombel J F , Takeuchi K , Gao X , Panaccione R , Danese S , Dubinsky M , Schreiber S , Ilo D , Finney-Hayward T , Zhou W , Phillips C , Yao X , Zhou Q , Loftus E
Published: January 2022

Background

Upadacitinib (UPA), an oral, reversible JAK inhibitor engineered for increased selectivity for JAK1 over JAK2, JAK3, or tyrosine kinase 2 (TYK2), demonstrated significantly greater efficacy compared with placebo (PBO) for induction of remission in patients with moderately to severely active ulcerative colitis (UC) in two phase 3 trials, U-ACHIEVE Induction (NCT02819635) and U-ACCOMPLISH (NCT03653026). This analysis evaluated the efficacy of UPA on early symptomatic improvement for the first 14 days, using pooled data from U-ACHIEVE Induction and U-ACCOMPLISH.

Methods

U-ACHIEVE and U-ACCOMPLISH were multicentre, double-blind, PBO-controlled trials that enrolled patients who have had moderately to severely UC with an Adapted Mayo Score of 5 to 9 points and centrally reviewed endoscopy subscore of 2 to 3. A total of 998 patients were randomized to receive UPA 45mg once daily (QD) (n=658) or PBO (n=328) for 8 weeks (wks) in a 2:1 ratio. First dose of study drug was administered on Day 0. Improvement in symptoms including stool frequency subscore (SFS), rectal bleeding subscore (RBS), abdominal pain, and bowel urgency were analysed from daily symptom diary data. Multivariate regression analysis was used to determine if early changes in UC symptoms could be used to evaluate a potential correlation with patients’ likelihood of achieving clinical response or clinical remission per Adapted Mayo score at the end of induction.

Results

Baseline characteristics were similar between both treatment groups. Patients treated with UPA 45 mg QD experienced significant improvement in daily symptoms, with significantly more subjects achieving SFS≤1 (p<0.001), RBS of 0 (p<0.05), and SFS of 0 (p<0.05) as early as day 1 and maintained through day 14 (Figure 1). A significantly higher percentage of patients who received UPA 45 mg QD compared to PBO, achieved abdominal pain=0 and the absence of bowel urgency within 3 days of beginning treatment through day 14 (p<0.05). Multivariate analysis revealed that patients who achieved day 7 SFS≤1 (OR 2.42, 95% CI, 1.53–3.82) were more likely to attain clinical response (Table). Patients who attained day 7 SFS≤1 (OR 2.53, 95% CI 1.59–4.00) or day 7 bowel urgency absent (OR 2.40, 95% CI 1.52–3.79) were more likely to achieve clinical remission at week 8.

Conclusion

UPA 45 mg QD significantly improved UC symptoms as early as day 1, providing patients with rapid symptom relief. Patients who achieved early symptom improvement were more likely to attain clinical remission or clinical response at week 8. [Clinicaltrials.gov, U-ACHIEVE Induction (NCT02819635) and U-ACCOMPLISH (NCT03653026)]
DOP56 Histologic disease activity correlates with endoscopic severity in patients with moderate to severe Crohn’s Disease
Authors: Reinisch W , De Hertogh G , Protic M , Chan L S , Magro F , Pollack P , Feagan B G , Harpaz N , Pai R
Published: January 2022

Background

The role of histopathology evaluation in Crohn’s disease (CD) is not precisely defined. Due to the heterogeneity of phenotypes, the discontinuity of distribution, and the transmural nature of inflammation, the interpretation of histologic outcomes is complex. To analyze how histology relates to established outcomes in CD, we examined association between histologic and endoscopic disease activity measured by frequently used scores (simple endoscopic score for Crohn’s disease [SES-CD], Robarts histopathology index [RHI], global histological activity score [GHAS]) and the association between histologic or endoscopic severity with inflammatory biomarkers in a cohort of patients with moderate-to-severe CD.

Methods

Patients (N=191) who were enrolled in a phase 2 randomized clinical trial (NCT02891226) in patients with moderate-to-severe CD were assessed at baseline (BL) for endoscopic disease location and severity as measured by SES-CD. Biopsies were obtained during BL endoscopy from the edge of the ulcers and the most inflamed mucosa in terminal ileum and 4 colonic segments (ascending, transverse, descending, rectum; N=10/patient, 2 per location with the more severe score used), and scored by central readers blind to study treatment, timepoint, and response status using RHI and both modified and active GHAS (mGHAS: Q1, 3, 4, 5, 6; aGHAS: Q1, 4, 5, 6; see Fig. 1). Inflammatory biomarkers included fCLP and CRP; log-transformed values are used. Linear correlations between measures were determined using Pearson correlation coefficients (PCC) as exploratory analyses. Nominal p values are presented.

Results

SES-CD correlated well with all 3 histologic measures examined (Fig. 1). While both fCLP and CRP correlated more closely with endoscopy than with histologic measures, fCLP had higher correlation with each histologic measure than CRP (Fig. 2). 64% of patients had the same disease location at BL when measured by endoscopy or histology. Patients with endoscopic colonic disease displayed histologic ileal involvement in up to 20% of cases, while 13% of patients with endoscopic ileal disease demonstrated histologic colonic involvement (Table 1).

Conclusion

Although histologic disease activity in CD, as determined by 3 different measures, correlates well with endoscopic assessment of disease, histologic involvement provides complementary information on disease distribution and extension. Further analyses are required to understand the additive role of mucosal histology in CD.
DOP68 CD-TREAT diet induces remission and improves quality of life in an open label trial in children and adults with active Crohn’s Disease
Authors: Svolos V , Hansen R , Russell R , Gaya D R , John Paul S , Macdonald J , Nichols B , Papadopoulou R , Logan M , Mckirdy S , Gervais R , Ijaz U , Milling S , Shields S , Wilson D , Henderson P , Din S , Ho G T , Gerasimidis K
Published: January 2022

Background

Exclusive enteral nutrition (EEN) is an established induction treatment for active Crohn’s disease (CD) with a proposed mechanism of action involving the gut microbiome. We have previously shown that CD-TREAT diet, a food-based diet with similar dietary profile to EEN, improves rat ileitis and replicates the effect of EEN on the gut microbiome of healthy volunteers and animal models. Here, we test the efficacy of CD-TREAT diet to induce clinical remission in active CD.

Methods

This is an open-label study in children (wPCDAI≥12.5) and adults (HBI≥5) with active CD. Primary outcome was clinical response (wPCDAI fall≥17.5; HBI fall≥3) or clinical remission (wPCDAI< 12.5; HBI<5) after 8 week treatment with CD-TREAT. Secondary outcomes included improvement of quality of life (QoL) and reduction in faecal calprotectin (FC) levels. Since CD-TREAT diet is gluten-free, adherence to treatment was assessed by the detection of the gluten immunogenic peptide (GIP) in faeces. Data are presented with median (IQR).

Results

25 children, [age, 14.4 (12.5,15.7) years] and 32 adults, [age, 32.6 (24.2,43.9) years] were treated. 7 (12%) failed treatment and n=10 (18%) dropped out during the first 2 weeks of treatment due to palatability issues. In patients who completed 8 weeks of CD-TREAT course (n=40), 85% and 78% achieved clinical response and remission, respectively. CD-TREAT diet improved QoL in children [IMPACT-III score, baseline: 136 (122,143) vs 8weeks: 148 (133,153), p<0.01] and in adults [sIBDq score, baseline: 30 (26,45) vs 8weeks: 60 (48, 64), p<0.001]. Faecal GIP decreased during treatment [ng/g stool, baseline: 1250 (589, 1250), 4weeks: 0 (0,269), 8weeks: 0 (0,329), mg/mg, p<0.001 for both] showing adherence with the CD-TREAT diet. However, 33% and 40% of the patients had detectable faecal GIP at 4 and 8 weeks, respectively, revealing at least partial non-adherence. 30% of patients who completed CD-TREAT (n=12/40) experienced >50% FC reduction. Median FC levels decreased significantly? in the group of patients (n=22) who had undetectable GIP at 4 or 8 weeks [mg/kg FC, baseline: 1190 (361,1129); 8weeks: 534 (92,1230), p<0.01].

Conclusion

CD-TREAT diet improved disease activity indices and QoL in the majority of patients who completed treatment and decreased FC in those who were most likely to be compliant. Future RCT should aim to compare CD-TREAT with other induction treatments and improve meal variety and palatability to improve compliance and reduce drop-out rates.
DOP87 Association of histologic measurement with endoscopic outcomes after one year of treatment with mirikizumab in patients with moderate to severe Crohn’s Disease
Authors: Magro F , De Hertogh G , Protic M , Chan L S , Pollack P , Feagan B G , Harpaz N , Pai R , Reinisch W
Published: January 2022

Background

There is a scarcity of data regarding the role of histopathology evaluation in Crohn’s disease (CD) and how it relates to endoscopy. Mirikizumab (miri) has shown efficacy in Ph2 studies in CD and ulcerative colitis (UC). Here we explore the agreement of histologic response and remission with endoscopic outcomes after 12 and 52 weeks of treatment with miri in a phase 2 randomized clinical trial (SERENTIY; NCT02891226) in patients with moderate-to-severe CD.

Methods

Patients were randomized 2:1:1:2 across 4 treatment arms (PBO, 200, 600, 1000mg miri) administered intravenously (IV) every four weeks (Q4W) at Weeks 0, 4, and 8. Those who received miri and achieved ≥1 point improvement from baseline (BL) at W12 in Simple Endoscopic Score for Crohn’s Disease (SES-CD) were re-randomized 1:1 into double-blind maintenance to continue their IV dose assignment Q4W (IV/IV) or to 300mg miri SC Q4W (IV/SC); due to small numbers, maintenance IV arms were pooled for analysis. W12 non-improvers and all induction PBO patients received 1000mg miri Q4W from W12 through W52. Biopsies were obtained during endoscopy at W0, 12 and 52 from the edge of the ulcers and the most inflamed mucosa in terminal ileum and 4 colonic segments (ascending, transverse, descending, rectum; N=10/patient, 2 per location with the more severe score used), and scored by central readers blind to study treatment, timepoint, and response status. Histologic endpoints were defined post-hoc but prior to performing the analyses (see Table footnotes for definitions). Cohen’s Kappa Coefficient was used to evaluate the degree of agreement of two measures.

Results

At Week 12, there was an overall 69.6% agreement in histologic and endoscopic response, and 84.2% agreement between histologic remission and endoscopic remission (Table 1). After 52 weeks of miri treatment, the agreement between histologic and endoscopic response was 61.9%, while the agreement between histologic and endoscopic remission was 70.6% (Table 2). Interestingly, in every comparison, the percentage of patients achieving the histologic outcome but not the endoscopic outcome was greater than that of patients achieving the endoscopic outcome but not the histologic outcome.

Conclusion

Histologic and endoscopic outcomes demonstrated fair association after both 12 and 52 weeks of miri treatment. Histologic endpoints, despite the stringent criteria, appear to be more sensitive to change than endoscopic endpoints. However, variability of biopsy sampling may be an additional factor contributing to these differences.
P002 Glial cell line-Derived Neurotrophic Factor (GDNF) improves intestinal wound healing – A new target for IBD treatment?
Authors: Kelm M , Burkard N , Hoerner M , Germer C T , Schlegel N , Flemming S
Published: January 2022

Background

Due to the rising incidence and socioeconomic relevance of Inflammatory Bowel Disease (IBD) worldwide, new therapeutic strategies are necessary to improve patient care. Since immunosuppressants are partially insufficient with relevant rates of side effects, novel concepts of treatment need to be developed. Mucosal and/or histological healing represent potential criteria to measure disease activity and are mentioned in current guidelines as favorable prognostic factors regarding disease outcomes. In previous studies, the soluble factor Glial cell line-Derived Neurotrophic Factor (GDNF) has been shown to be significantly reduced in tissue specimens from IBD patients and to be critically involved in intestinal epithelial barrier maturation. Therefore, the goal of this study was to further analyze the potential role of GDNF in IBD.

Methods

Intestinal wound healing was assessed in endoscopic biopsy-based wound assays as well as dextran sodium sulphate (DSS)-induced colitis in vivo. During the experiments, C57Bl/6 mice were either injected with GDNF or sodium chloride intraperitoneal while tissue samples were used for detailed molecular analyses. Further, scratch wound assays were performed in Caco2 monolayers to identify molecular pathways and cell mechanisms in vitro.

Results

Intraperitoneal injection of GDNF resulted in significantly enhanced wound closure in vivo. During the DSS-induced colitis model, mice treated with GDNF demonstrated significantly improved recovery from colitis in comparison to mice injected with sodium chloride. Similarly, application of GDNF in a scratch wound assay significantly enhanced wound closure in vitro. Mechanistically, molecular analysis revealed that GDNF application resulted in significantly increased cell proliferation. The effect of GDNF on cell proliferation was based on upregulation of LGR5-positive cells with increased phosphorylation of pSrc. Further studies demonstrated that the effect of GDNF on wound healing and cell proliferation could be inhibited by application of a Src-inhibitor.

Conclusion

GDNF significantly improves intestinal mucosal healing in vivo and in vitro which results in significantly enhanced recovery from colitis. While this effect is driven by increased cell proliferation due to upregulation of LGR5-positive cells and increased phosphorylation of pSrc, previous studies already demonstrated the critical impact of GDNF on epithelial barrier function. Following that, GDNF might have great potential in IBD treatment and could be a new target for future therapeutic approaches.
P004 Microbiota, not host origin drives ex vivo epithelial response in ulcerative colitis patients and non-IBD controls
Authors: Arnauts K , Sudhakar P , Verstockt S , Lapierre C , Potche S , Caenepeel C , Verstockt B , Raes J , Vermeire S , Sabino J , Verfaillie C , Ferrante M
Published: January 2022

Background

Host-microbial interactions in inflammatory bowel disease (IBD) are poorly understood. Furthermore, patients with IBD have microbial dysbiosis and their epithelial cells exhibit intrinsic defects. We aimed to unravel the effect of exposure to microbiota on epithelial cells from both patients with ulcerative colitis (UC) and non-IBD controls.

Methods

Confluent Transwell® organoid-derived monolayers of 8 UC patients and 8 non-IBD controls were co-cultured for 6 hours with microbiota (3.108 cells) derived from active UC patients (n=3, endoscopic Mayo score ≥2) or healthy volunteer (HV, n=1, selected on high microbial cell count and presence of selected phyla1). For this purpose, fresh faecal samples were filtered and frozen in 0.9% NaCl. Inflammation was re-induced with 100 ng/ml TNF-α, 20 ng/ml IL-1β, and 1 µg/ml flagellin in confluent Transwell® cultures, 24 hours prior to microbiota co-culture. Transepithelial electrical resistance (TEER), 4 kDa fluorescein isothiocyanate (FITC) dextran (2 mg/ml) measurements, and RNA sequencing by Truseq were performed on epithelial cells, and 16S rRNA sequencing on microbiota samples before and after co-culture.

Results

The transcriptomic response of epithelial cells to microbiota was clearly influenced by the type of microbiota stimulation (no, UC or HV microbiota). However, this response was not influenced by the origin of the epithelial cells (UC or non-IBD controls) (Figure 1A). TEER and FITC dextran measurements showed a strong decrease in epithelial integrity (in both UC and non-IBD epithelium) following stimulation with UC microbiota, in contrast to HV microbiota (Figure 1B). In UC epithelial cells, stimulation with microbiota from UC patients induced a stress induced phenotype including activation of EGR1 signalling, AP-1 family, FOSL genes, MAPK and JNK signalling (Figure 2). Activation of stress pathways (MAPK family cascades) and key markers (e.g. FOSB, FOSL1, MYC, EGR1, ATF3, NOTCH1) was lower after HV microbiota stimulation (Figure 3–4). Expression levels of key markers were UC microbiota specific and similar in epithelial cells of UC and non-IBD controls (Figure 4).

Conclusion

Not the host (UC vs non-IBD epithelial cells) but the microbial donor (UC vs HV) is driving the transcriptomic response. Epithelial cells from UC patients did not show an increased sensitivity towards microbiota stimulation compared to non-IBD epithelial cells. In contrast, exposure of epithelial cells to UC microbiota was sufficient to induce a strong stress response and barrier disruption. Further research on therapies to restore the microbial balance, to remove the constant trigger of dysbiosed microbiota, is required.

References

1. Vermeire, JCC, 2015
P007 ORMDL proteins shape homeostasis in the intestinal epithelium by regulating endoplasmic reticulum architecture and autophagy
Authors: Tran F , Stengel S , Yang H , Bernardes J P , Lopez-Agudelo V , Bordoni D , Falk-Paulsen M , Jentzsch M , Messner B , Schreiber S , Rosenstiel P
Published: January 2022

Background

ORM1-like protein 3 (ORMDL3) is an endoplasmic reticulum (ER)-resident protein that has been implicated in ER homeostasis and sphingolipid biosynthesis. Genome-wide association studies identified susceptibility loci around the ORMDL3 gene for inflammatory diseases, including asthma and IBD. Preliminary studies suggested a role of ORMDL proteins within the ER stress-induced unfolded protein response (UPR). Here, we set out to investigate the role of ORMDL proteins in intestinal inflammation.

Methods

Mice with constitutive and conditional knockouts (KO) of Ormdl1, Ormdl2 and Ormdl3 in intestinal epithelial cells were generated. From these, intestinal organoids were generated to perform functional assessment of ER stress, autophagy, and inflammatory responses. We performed 3’-single cell RNA sequencing (scRNAseq) of small intestine tissue of animals with triple conditional knockout of Ormdl1/Ormdl2/Ormdl3 (“Ormdl123-VilCre”) vs. wildtype to identify cell type-specific effects of epithelial ablation of ORMDL proteins. 16S rDNA sequencing was performed on stool samples from these animals.

Results

Mice deficient for ORMDL displayed reduced body weight dependent on the number of knockouts. Ormdl1/2/3-KO mice are embryonically lethal while Ormdl1/2/3-VilCre animals were viable without signs of spontaneous inflammation. Ormdl1- and Ormdl3-KO animals displayed reduced number of Paneth and goblet cells in the small intestine, accompanied by massive dilation/ballooning of the ER. This phenotype was strongly pronounced in Ormdl1/3-KO and Ormdl1/2/3-VilCre animals. ORMDL3 co-localises and co-precipitates with LC3, indicating a direct interaction with autophagy proteins, while loss of ORMDL proteins impaired autophagic flux. Interestingly, UPR and Cxcl1 expression were insufficiently induced in intestinal organoids from Ormdl1/2/3-VilCre animals treated with tunicamycin. scRNAseq analysis of small intestine tissue surprisingly revealed compensatory induction of antimicrobial peptides in non-Paneth cells and vice versa downregulation of apoptosis regulating gene in Ormdl1/2/3-VilCre mice. We observed an expansion of myeloid and plasma cells in Ormdl1/2/3-VilCre mice, while no changes in epithelial cell proportions can be identified. These findings were accompanied by dysbiosis of the stool microbiome (β-diversity, Bray-Curtis dissimilarity) of Ormdl1/2/3-VilCre animals.

Conclusion

Our data suggest a critical regulatory role of the IBD risk gene ORMDL3 in the ER-autophagy axis in intestinal epithelial cells, shaping the mucosa homeostasis on cellular and microbiome level. Functional assessment of inflammation in Ormdl1/2/3-VilCre animals (e.g., DSS colitis model) will be performed to understand the role of ORMDL in intestinal inflammation.
P012 The effect of Oncostatin M on human intestinal organoids
Authors: Filidou E , Kandilogiannakis L , Kokkotis G , Tarapatzi G , Arvanitidis K , Drygiannakis I , Valatas V , Paspaliaris V , Kolios G , Bamias G
Published: January 2022

Background

We have previously shown that human subepithelial myofibroblasts (SEMFs), which are of mesenchymal origin, express the receptor of Oncostatin M (OSM), which is further induced when SEMFs are previously exposed to IL-1α and TNF-α. Human Intestinal Organoids (HIOs), derived from embryonic stem cells (ESC), form epithelial crypts consisting of several subtypes of epithelial cells and are surrounded by cells of mesenchymal origin. Recently, we reported that the mesenchymal component of HIOs is gradually reduced, as culture passages increase. The aim of our study was to examine the effect of exogenous OSM on fibrotic and pro-inflammatory marker expression of HIOs, with and without the combined presence of IL-1α and TNF-α.

Methods

The human ESC line (H1)-derived HIOs were developed using a commercially available kit and characterized by immunofluorescence in all differentiation stages. HIOs from passage 2 were stimulated with either 100ng/ml OSM for 12 hours or 5ng/ml IL-1α and 50ng/ml TNF-α for 24 hours and then with 100ng/ml OSM for 12 hours. mRNA expression of fibrotic markers, Collagen Type I, III, and Fibronectin, and pro-inflammatory markers, CCL2, CXCL10 and CXCL11 were examined by reverse transcription quantitative PCR.

Results

OSM alone significantly downregulated both Collagen Type I and III mRNA expression, and had no effect on Fibronectin expression. When HIOs were previously exposed to IL-1α and TNF-α, however, OSM stimulation resulted in significant upregulation of Collagen Type I and Fibronectin, and had no effect on Collagen Type III. Regarding the pro-inflammatory marker expression, OSM alone induced the expression of CCL2, CXCL10 and CXCL11, with a greater effect observed when HIOs were previously exposed to IL-1α and TNF-α.

Conclusion

Our findings indicate that OSM induces the expression of both fibrotic and pro-inflammatory factors in early passages of HIOs, previously exposed to IL-1a and TNF-a, as has been previously observed on SEMFs. Since the mesenchymal cell component of HIOs is strongly present in early passages and OSM’s receptor is mainly found to be expressed in these cells, our results suggest OSM affects HIOs via the cells that are of mesenchymal origin.
P015 Detection of endoscopic lesions from limited quality annotations in colonoscopy videos
Authors: Alali S , Chaussard J , Li-Thiao-Té S , Ogier-Denis É , Percy-du-sert A , Treton X , Zaag H
Published: January 2022

Background

Ulcerative colitis is an idiopathic inflammatory disorder affecting the mucosa of the colon with superficial erosion and ulcers associated with bleeding. Severity assessment using current scoring schemes such as UCEIS and MAYO relies on the subjective interpretation of the physician and fails to take into account the size of the lesions, their number and distribution. Automatic lesion detection methods can enable fine-grained assessment of lesion severity, but require training stage based on time-consuming manual annotation. Most methods currently use generic datasets that are biased towards capsule endoscopy and are not adapted to locally available hardware.

Methods

We learn automatic bleeding and ulcer detectors on a local data set created by the Gastroenterology group at the Bichat and Beaujon hospitals. The patients’ videos were anonymous, analysed after obtaining their consent. The study was approved by the local research study committee. To minimise expert annotation burden, only rectangular annotations are provided instead of a precise delineation of lesion boundaries (Figure 1). This leads to many mislabelled pixels, especially in the corners, and affects the evaluation of the models’ performance and our ability to find correct models. Standard sensitivity and specificity cannot be used effectively on this data-set. We propose to evaluate model sensitivity on the annotation level and keep specificity at the pixel level. On the training set, we consider that a model correctly identifies a lesion if it agrees with the expert on a subset of the annotation, and count the detected annotations weighted by their area. For robustness and ease of interpretation, we explore the set of linear classifiers, and propose an efficient sampling scheme that rejects trivial models. This method is evaluated on a database of 10 colonoscopy videos (5 training videos and 5 test videos).

Results

In spite of the limited quality of the annotations, we find lesion detectors with a good annotation-level sensitivity (Table 1) and visual performance (see Figure 1). The detector’s performance is computed reliably. We evaluate sensitivity and specificity on 20 random subsets containing 10\% of the images, and obtain similar performance for the same patient for all models (cf Figure 2).

Conclusion

Despite mislabeled pixels, we obtain lesion detectors with good performance, and we show that the performance is computed reliably. However, the inter-patient performance is variable and the best models fail on some patients (sensitivity below 20\% in some cases Figure 2). This suggests that the models are not universal and that the appearance of bleeding and ulcers should be normalised further before automatic detection.
P017 Transcriptional characterization of lymphocytic colitis and comparison to collagenous colitis
Authors: Koch S , Münch A , Escudero-Hernández C
Published: January 2022

Background

Lymphocytic colitis (LC) is an inflammatory disease of the large intestine that causes chronic, watery, non-bloody diarrhoea. LC is characterized by infiltration of lymphocytes in the colonic mucosa. To better understand LC pathophysiology on a molecular level, we performed genome-wide RNA sequencing (RNA-seq) analysis on LC mucosal samples.

Methods

We collected colonic biopsies from healthy controls (Hc), and LC patients with active disease. Collagenous colitis (CC) samples were included as a separate control. Total RNA was isolated from biopsies for bulk RNA-seq. Whole-genome expression data were analysed by principal component analysis (PCA), differential gene expression (DGE), and gene-set variation analysis (GSVA) using R statistical software. Benjamini-Hochberg FDR was used to adjust p values, considering <0.05 as statistically significant. Gene-set enrichment analysis (GSEA) was performed using GSEA software and visualized in Cytoscape.

Results

PCA of all samples identified 2 principal components that explained 65% of the transcriptional variation and separated sample groups into distinct clusters of gene expression. Subsequent analysis revealed differential expression of 5045 genes in patients with active LC compared to Hc, which were mainly involved in DNA and telomere repair, in ubiquitinylation and proteasomal degradation, response to viruses and interleukin (IL)-12 signalling according to GSEA. Compared to CC samples, we identified 384 differentially expressed genes (DEGs) with LC. Pathway analysis displayed an enrichment in cell adhesion and proliferation genes in LC, whereas DEGs related to microbial response, collagen and wound healing processes, chemotaxis, and epithelial biology (e.g. ion channels, epithelial cell proliferation) were enriched for CC samples. Estimations of infiltrating immune cells performed by GSVA indicated a possible role for professional antigen-presenting cells (i.e. macrophages and inactive dendritic cells) as well as T regulatory cells in LC pathogenesis.

Conclusion

LC is a transcriptionally homogeneous disease that can be characterised by increased expression of cell adhesion and proliferation-related genes. Furthermore, our analyses confirm an elevated immune response in LC, particularly involving macrophages, dendritic and T regulatory cells. Comparison of LC and CC gene expression additionally supports that these diseases are pathobiologically distinct.
P018 Proteomic profile of serum and urine in newly diagnosed patients with Inflammatory Bowel Disease: new approach for biomarker discovery
Authors: Baldan-Martin M , Azkargorta M , Iloro I , Ortega Moreno L , Aldars Garcia L , Soleto Fernández I , Ramirez C , Riestra S , Rivero M , Gutierrez A , Rodríguez-Lago I , Fernández-Salazar L , Ceballos D , Benítez J M , Aguas M , Bastón-Rey I , Bermejo F , Casanova M J , Llorente R , Ber Y , Royo V , Esteve M , Elortza F , Chaparro M , Gisbert J P
Published: January 2022

Background

Currently, endoscopy is the gold standard for the evaluation of disease activity and inflammation in clinical practice. However, it is an invasive procedure for the patient, costly and time-consuming. Therefore, there is an urgent need to identify non-invasive biomarkers with the potential clinical utility to diagnose inflammatory bowel disease (IBD) patients, improve the patient′s quality of life and increase efficiency in the Health Systems.

Methods

A label-free quantitative proteomics method was used to profile the urinary and serum proteomes of 100 patients with newly diagnosed IBD, before starting any treatment [50 patients with Crohn′s disease (CD) and 50 patients with ulcerative colitis (UC)] and 50 healthy controls (HC). The resulting peptides were separated by NanoLC (EVOSEP ONE) and coupled online by electrospray to Trapped Ion Mobility Spectrometry (TIMS) TOF Pro for tandem mass spectrometry analysis. Mass spectrometry data were processed and analyzed with MaxQuant and Perseus software. Proteins with p≤0.05 and ratio>1.5 in any sense, were considered as significantly dysregulated. Bioinformatic analysis of differentially expressed proteins was performed to characterize involved biological processes, molecular functions, cell locations and proteins pathways (PANTHER and STRING v11.5).

Results

A total of 2,500 proteins were identified in the urine, and 468 proteins were identified in the serum with at least two different peptides at FDR<1%. Of the urine proteins, 110 were significantly changed in UC versus HC, 50 proteins were differentially expressed between CD and HC, and a total of 31 proteins were significantly changed in CD compared with UC patients (Figure 1). In serum samples, a total of 45 differentially expressed proteins were identified in the comparison between UC and HC groups, 32 proteins significantly expressed in CD versus HC, and 12 proteins in CD compared with UC (Figure 2). Finally, biological information analysis was performed according to these differential proteins. The molecular functions, biological processes, and pathways enriched mainly involved the neutrophil degranulation, complement and coagulation cascades and immune system responses (Figures 3 and 4).

Conclusion

Proteome profiling revealed significantly differences in newly diagnosed patients with CD or UC. Both serum and urine seem to be appropriate biological matrixes for this approach. Next steps of our research will be to understand the role of candidate proteins as well as to validate those with potential as biomarkers in independent cohort.
P021 Vitamin D receptor-dependent protective effect of moderate hypoxia in a mouse colitis model Running title: Hypoxia prevents mouse colitis through VDR
Authors: Wang Z , Hong Y , Hong L , Changzhi H , Jiaming Q
Published: January 2022

Background

Although physiological hypoxia is important for maintaining the intestinal barrier, its effect on the barrier during acute colitis and the underlying mechanisms are not fully understood. Here,we aim to explore the influence of hypoxia in dextran sulfate sodium (DSS)-induced colitis mice and the role of hypoxia-inducible factor (HIF) and vitamin D receptor (VDR) in the process

Methods

Colitis mice were subjected to hypoxia to detect colitis symptoms and intestinal barrier function changes. Finally, the mechanisms underlying the changes were explored in a colon cell line.

Results

First, compared with colitis mice without hypoxia stimulation, those with hypoxia stimulation showed significantly decreased pathological damage and improved permeability of the intestinal barrier. The expression of tight junction proteins (including occludin, claudin-1 and ZO-1), HIF-1α as well as VDR was up-regulated in colitis mice with hypoxia stimulation. However, in VDR gene knockout (VDR-KO) colitis mice, hypoxia treatment showed no protective effect, suggesting the VDR dependency of this effect. Second, we identified that although hypoxia stimulation could enhance the single-layer epithelial transmembrane electrical resistance in DLD-1 cells, these effects disappeared in VDR-knockdown cells. Furthermore, over-expression of HIF-1α in DLD-1 can increase the expression of VDR, then chromatin immunoprecipitation and luciferase assays confirmed that HIF-1α can bind to the promoter region of the VDR gene under hypoxia. Finally, compared with their wild-type siblings, VDR-KO mice showed reduced abundance of anaerobic bacteria and SCFA-producing bacteria.

Conclusion

Moderate hypoxia was protective against DSS-induced colitis, and VDR is instrumental in it. Furthermore, HIF-1α mediates the effect of hypoxia on the VDR signaling pathway. Moreover, intestinal flora may be an important link between hypoxia and VDR.
P022 Transcriptional biomarkers for vedolizumab therapy response in patients with moderate to severe Ulcerative Colitis
Authors: Arosa García L , Camba-Gómez M , Calviño-Suárez C , Bastón-Rey I , Ferreiro-Iglesias R , Porto M , Nieto-García L , Domínguez-Muñoz J E , Barreiro-de Acosta M , Conde-Aranda J
Published: January 2022

Background

Vedolizumab is one of the current treatments for patients with Inflammatory Bowel Disease (IBD). The efficacy and safety, together with its gut specificity, make this drug an appealing therapeutic option for IBD patients with moderate to severe disease. However, as observed for other biologic treatments, a significant proportion of patients do not have an initial response to vedolizumab treatment. Currently, there is a lack of reliable biomarkers for vedolizumab treatment response, although this would help to palliate the socioeconomic costs derived from this disease. For that reason, the primary aim of this study is to establish the basis for the search of transcriptional factors associated with vedolizumab treatment response.

Methods

For the realization of this study, we collected blood samples from responder and non-responder Ulcerative Colitis (UC) patients treated with vedolizumab. The clinical response was measured using the Partial Mayo Score. The frequencies of different immune system populations were analysed by flow cytometry. Moreover, we measured the transcriptional levels of several genes in peripheral blood mononuclear cells (PBMC) by RT-qPCR. These experimental procedures were performed at baseline (T0) and after 14 weeks of follow-up (T14).

Results

We enrolled nine patients with an average age of 46.25±11.71, of which seven were previously treated with anti-TNF therapy. Our results show a specific pattern in responder and non-responder patients in the percentages of different innate and adaptive immune cell populations at T0 versus T14. Similarly, we observed a significant reduction in the expression of some chemokines (i.e., CCL25) and pro-resolutive factors (i.e., ANNEXIN A1) at T14 versus T0 in PBMCs from vedolizumab responder patients, which was not observed in non-responder patients.

Conclusion

Our data suggest that the frequencies of certain immune populations are associated with the response to vedolizumab treatment. In the same way, we found that specific transcripts are modulated in response to this α4β7 integrin antibodies in PBMCs. Therefore, we found a solid system to search for vedolizumab therapy response markers using low invasive techniques.