bluuebunny/crossref_metadata_2025 huggingface dataset:
DOP46 Metabolic adaptation to ER stress licences STING signalling in intestinal epithelial cells
Authors: Wottawa F , Becker B , Bakr M , Kugler J , Mayr L , Paludan S , Blumberg R , Kaser A , Adolph T , Schreiber S , Meiser J , Rosenstiel P , Aden K
Published: January 2022

Background

Chronic endoplasmic reticulum stress (ER) in the intestinal epithelium is a pathophysiological hallmark of IBD. cGAS/STING is an innate immune pathway involved in the detection of double stranded DNA fragments leading to the subsequent induction of type I IFN responses. We here tested the hypothesis that chronic ER stress impairs cGAS/STING signalling in the intestinal epithelium.

Methods

Mice with a conditional intestinal epithelial deletion of Xbp1 (Xbp1 ΔIEC, Xbp1fl/fl) were used to assess intestinal epithelial STING expression in-vivo. Small intestinal organoids (Xbp1ΔIEC, Xbp1fl/fl) and cell lines (Mode K, iCtrl and iXbp1) were used to assess cGAS/STING signalling in-vitro using STING agonist (dsDNA, DMXAA). Murine cytomegalovirus (mCMV) infection assays were performed in iCtrl and iXbp1cells and Xbp1ΔIEC, Xbp1fl/fl mice to functionally link impaired cGAS/STING to pathogen response. LC-MS profiling was performed in iCtrl and iXbp1cells to identify underlying metabolic programs affecting cGAS/STING responses in ER-stressed cells. IBD biopsy samples (cross-sectional, longitudinal therapy response cohort) were used to validate key molecular phenotypes in human IBD.

Results

Compared to Xbp1 fl/fl mice, Xbp1ΔIEC show completely abrogated STING expression in the basal crypt compartment of the small intestinal epithelium. In line with that iXbp1 ModeK cells displayed impaired pathway activation (TBK1) and interferon inducible gene expression (Cxcl10) in response to cGAS/STING stimulation and towards mCMV infection, leading to increased viral replication compared to iCtrl cells. In-vivo mCMV infection led to augmented small intestinal histopathological disease activity in Xbp1ΔIEC, but not Xbp1fl/fl mice. Using LC-MS, we show that ER-stress induces a metabolic adaptation towards increased serin/glycin metabolism, which is used to counterbalance reactive oxygen species (ROS) via glutathione (GSH) synthesis. Pharmacological interception of key pathways of GSH synthesis of deprivation of serin/glycin phenocopies ER-stress in abrogating STING signalling in IECs. Lastly, we show that key aspects of metabolic adaptation to ER-stress are present in intestinal biopsies of IBD patients.

Conclusion

Our data describe a novel mechanism of metabolic adaptation to compensate ER-stress and maintain intestinal epithelial cGAS/STING signalling. We therefore put forward a model of ER-stress driven immunodeficiency via cGAS/STING signalling which renders the intestinal mucosa susceptible towards CMV infection in the context of IBD.
DOP48 Hedgehog signalling controls Th17 differentiation to drive intestinal inflammation and is a druggable target for the treatment of IBD
Authors: Hanna J , Beke F , O’Brien L , Kapeni C , Chen H C , Carbonaro V , Kim A , Kishore K , Adolph T E , Skjoedt M O , Skjoedt K , de la Roche M , de la Roche M
Published: January 2022

Background

T helper 17 (Th17) cells play an important role in barrier protection in the gastrointestinal tract but are also key pathological drivers of Inflammatory Bowel Disease (IBD). Although a number of transcription factors governing Th17 differentiation have been identified, the intracellular signalling pathways regulating Th17 differentiation are poorly understood. Hedgehog (Hh) signalling controls cell-fate choices in numerous tissue compartments and is targetable by highly selective, clinically-approved small molecule inhibitors. However the role of Hh signalling in Th17 differentiation and effector function is unstudied.

Methods

We generated two conditional knockout mouse models targeting Hh signalling components Smo and Ihh to study Th17 differentiation in vitro by flow cytometry and gene expression analysis. For in vivo studies, T cell adoptive transfer colitis was performed using donor Ihh knockout T cells or heterozygote controls. Histological analysis, mouse weight, colon length/weight measurements, and flow cytometric analysis was performed. We supplement this with the use of two small-molecule Smo antagonists for in vitro and in vivo studies of Th17 function. To underscore the translational relevance of our findings, we conducted bioinformatic analyses of published gene expression datasets of human rectal biopsies from two large independent cohorts of Ulcerative Colitis patients and healthy controls.

Results

We find that intracellular Hh signalling, independently of extracellular Hh ligands, selectively drives differentiation and effector function of Th17 cells but not of other T helper cell lineages. We demonstrate in vivo that inhibition of the Hh pathway with either the clinically-approved small molecule inhibitor vismodegib or genetic ablation of Ihh in CD4+ T cells results in a significant decrease in histological and clinical readouts of disease severity as well as a significant reduction in IL-17a+ Th17 cells. Our bioinformatic analyses show that Hh component expression levels are upregulated in human Ulcerative Colitis patient samples and are closely correlated with expression of Th17 markers. Mechanistically we show that the T-cell-intrinsic Indian Hedgehog (Ihh) ligand signals via the signal transducer Smoothened to activate both canonical and non-canonical Hh pathways, through the Gli3 transcription factor and AMPK phosphorylation, respectively.

Conclusion

We uncover Hh signalling as a novel pathway controlling Th17 differentiation and pathogenicity in IBD with Gli3 acting as a newly-identified crucial regulatory transcription factor. Our work paves the way for the use of Hh inhibitors for the treatment of IBD.
P006 MiR-192 is associated with stricturing phenotype in Crohn’s disease
Authors: Cristina T , Mateescu B , Mitroi A , Cozaru G , Brinzan C , State M , Dumitru A , Rafti R , Dina E , Alexandrescu L , Dumitru E
Published: January 2022

Background

MicroRNAs (miRNAs) are small, non-coding short (ribonucleic acid) RNAs that have a role in the regulation of genes and protein expression. MiRNAs have altered expression in multiple autoimmune disorders including inflammatory bowel diseases (IBD). This emerging role has led to investigations into miRNA expression profiles in IBD to understand the pathogenesis of these diseases and lead to clinical advances in this area. Due to shared pathways in fibrosis, miRNAs that have a potential role in intestinal fibrosis are worthy of investigation, and miR-192 seems to be associated with fibrosis and strictures in Crohn’s disease (CD). The study aimed to assess the expression levels of tissue and circulating miR-192 in patients with CD.

Methods

The study included 58 patients with CD and 31 subjects as controls for the validation cohort. Demographic and clinical data (including phenotype of the disease), biomarkers: fecal calprotectin (fCal), endoscopy data: Short Endoscopic Score of Crohn’s disease (SES-CD), the expression levels of miR-192 in tissue and serum were assessed (by RT-PCR). Receiver operating characteristic (ROC) analysis was performed to assess the miR-192 expression level as a potential biomarker for stricturing phenotype of CD.

Results

The mean fCal was 345+/- 112 μg/g. The mean SES-CD was 3.4 +/- 1.1. Regarding disease phenotype, 26 (44.8%) had inflammatory phenotype, 32 (55.2%) had stricturing phenotype, and no patient had penetrant phenotype. Regarding location, 20 (34.5%) were ileal, 5 (8.7%) were colonic, and 33 (56.8%) were ileocolic. Strictures were all located in the terminal ileum. The expression level of miR-192 was significantly upregulated in CD tissue samples compared to the control tissue samples only in the patients with stricturing phenotype (8.27 ± 2.87 vs. 4.20 ± 1.94; p < 0.01), whereas in inflammatory phenotype miR-192 was downregulated (-3.47 ± 1.02 vs. - 4.30 ± 0.84; p < 0.01). Similarly, serum miR-31 expression level in CD patients was significantly upregulated than in control serum samples only in the patients with stricturing phenotype (4.07 ± 1.00 vs. 1.55 ± 0.91; p < 0.01), whereas in inflammatory phenotype miR-192 was downregulated (- 0.25 ± 0.79 vs. 0.76 ± 0.83; p < 0.01). There was a strong correlation between tissue and circulating miR-192 with stricturing phenotype (r = 0.89, r = 0.85, p < 0.05). ROC analysis revealed that the expression level of miR-192 could help to discriminate stricturing CD patients from other phenotypes with very good specificity and sensitivity.

Conclusion

MiR-192 is associated with stricturing phenotype in CD and could be a biomarker of intestinal fibrosis. Furthermore, circulating miR-192 could be a non-invasive biomarker of intestinal fibrosis.
P033 Single-cell analysis of gut mucosal- and peripheral blood cells in ulcerative colitis patients undergoing vedolizumab treatment
Authors: Karmi N , Sun S , de Jong S , Bangma A , Teunis J , Paraskevopoulou M , Juarez J , Bleck B , Farahmand S , Weersma R , Bigaeva E , Festen E
Published: January 2022

Background

Vedolizumab (VDZ), a monoclonal antibody that targets α4β7 integrin, was approved to treat moderate-to-severe ulcerative colitis (UC) based on the presumption that it blocks T cell recruitment to the inflamed intestinal mucosa. The clinical evidence suggests that up to 50% of UC patients do not achieve disease remission under VDZ treatment. This study aims to identify changes in cell abundances and molecular pathways associated with VDZ response in UC. To this end, we included anti-tumor necrosis factor (anti-TNF)-naïve and anti-TNF-exposed patients with active UC, and utilized single-cell RNA sequencing (scRNAseq) and high-dimensional flow cytometry (Cytek) to assess the peripheral blood and the gut mucosal compartments.

Methods

Gut mucosal biopsies from inflamed and non-inflamed regions, and peripheral blood mononuclear cells (PBMCs) were obtained from UC patients 2 wks before (t0) and 14 wks after (t4) the start of VDZ administration. Response to treatment was prospectively evaluated based on endoscopic assessment (defined as a decrease in total Mayo score between t0 and t4) and physician global assessment (PGA) that incorporates disease activity score and biochemical measurements.

Results

A total of 25 UC patients (pts) were included: 44% anti-TNF-naïve. Endoscopic response to VDZ was observed in 32% of UC pts, while 56% of pts showed response based on PGA. The VDZ response rate (by PGA) was higher in anti-TNF-naïve pts vs anti-TNF-exposed pts (82% vs 36% responders, respectively). A preliminary analysis was performed on samples from 8 (out of 25) UC pts, profiling >70,000 gut mucosal cells and >25,000 PBMCs. Within the mucosal compartment, at t0 we identified immune cells (50% of all captured cells), stromal cells (10%), and epithelial cells (40%). Upon inflammation, the proportion of immune cells increased to 70%, stromal cells to 20%, while epithelial cells depleted to 10%. Notably, all main identified immune cell lineages – T cells, B cells and myeloid cells – contributed to the expansion of the immune cell compartment in inflamed mucosa. In line with scRNAseq data, we identified all major immune cell populations and detected expression of both the classic gut-directed and the redundant trafficking integrins by Cytek.

Conclusion

The preliminary results substantiate our current understanding of VDZ biology in UC. We confirm that anti-TNF-naïve pts have a higher response rate to VDZ vs anti-TNF-exposed pts. With this unique cohort, our study has the power to further explore molecular mechanisms and pathways that underlie VDZ response at the single-cell level.
P046 Effects of CB2 and TRPV1 stimulation on osteoclast overactivity induced by iron in pediatric inflammatory bowel disease
Authors: Strisciuglio C , Tortora C , Di Paola A , Creoli M , Cenni S , Argenziano M , Martinelli M , Miele E , Rossi F
Published: January 2022

Background

The reduction of bone mineral density and osteoporosis (OP) has an high impact on inflammatory bowel disease (IBD) patients’ health. We have previously shown that a dysregulated iron metabolism occurs in IBD and leads to a decrease in circulating iron concentration and an excessive intracellular sequestration of iron. Studies suggest that iron overload affects the bone, accelerating osteoclasts (OCs) differentiation and activation with consequent bone resorption. The aim of this study was to evaluate the role of Cannabinoid Receptor type 2 (CB2) and transient receptor potential vanilloid type-1 (TRPV1) and of the iron in the development of osteoporosis in pediatric IBD.

Methods

We enrolled 21 young subjects aged less than 18 years stratified into 3 groups on the basis of the clinical diagnosis: 7 with Ulcerative Colitis (UC) 7 with Crohn’s disease (CD) and 7 were non-IBD controls (CTR). We evaluated the role of CB2 and TRPV1 receptors and of iron in the development of OP in pediatric IBD at diagnosis and analyzed the effects of the pharmacological modulation of CB2 and TRPV1 receptors on osteoclast activity and on iron metabolism by Western Blotting, Tartrate-resistant acid phosphatase assay (TRAP assay) and Iron Assay Kit. We evaluated bone resorption through a commercially available kit visualizing and counting the resorption pits with the optical microscope.

Results

We observed higher levels of TRPV1 and lower levels of CB2 in the OCs from UC and CD patients compared to CTR. In IBD patients we found a significant up-regulation of osteoclast markers TRAP and Cathepsin K and a consequent osteoclast hyper-activation. The treatments of OCs from IBD patients with the CB2 agonist JWH-133 and the TRPV1 agonist Resinferatoxin, RTX, led to a significant reduction of osteoclastic hyper-activation affecting bone markers expression, number and size of active OCs, and bone resorption area. We also analyzed the [Fe3+] concentration and the expression of iron metabolism modulators, ferroportin1(FPN-1) and divalent metal transporter1(DMT1), in OCs derived from IBD patients. We demonstrated an increase of [Fe3+] and accordingly a decrease of FPN-1 in both groups of patients and an increase of DMT1 levels. After 48h of exposure to JWH-133 and RTX we observed a significant reduction of DMT1 expression in OCs from IBD patients together with a strong increase of FPN-1.

Conclusion

Our data confirm the well-known role of CB2 and TRPV1 receptors in bone metabolism and suggest that their stimulation can reduce the osteoclast overactivity induced by iron, thus providing new insights into the pathogenesis and in the treatment of pediatric IBD-related bone resorption.
P052 Comparing T cell responses to food antigens in Inflammatory Bowel Disease patients and healthy donors
Authors: Schiffer A , Tran F , Sievers L K , Kümpers J , Hinrichsen H , Seegers B , Bergfeld A , Lessing M , Schreiber S , Bacher P
Published: January 2022

Background

The role of nutrition in Inflammatory Bowel Disease (IBD) remains unclear. Studies about the effects of nutritional components on intestinal inflammation and the gut microbiome, as well as the concept of exclusive enteral nutrition in paediatric Crohn’s Disease (CD) are suggestive of an impact of nutrition on the pathophysiology. However, there is currently a lack of knowledge on how food components may affect IBD. Here, we investigated the T helper (Th) cell response against nutritional antigens in healthy controls and IBD patients.

Methods

We used a highly sensitive and specific method developed in our lab, the antigen reactive T cell enrichment (ARTE), to detect wheat and milk-reactive CD4+ T cells directly ex vivo in blood of healthy controls (n=18), CD (n=17) and Ulcerative Colitis (UC, n=15) patients. Peripheral blood mononuclear cells (PBMC) were incubated with whole antigen lysates and activated T helper cells were identified based on the up-regulation of the activation marker CD154. Magnetic enrichment of CD154 positive cells enabled to collect sufficient numbers of food-reactive T cells for their detailed characterization by multiparameter flow cytometry.

Results

We detected low frequencies of milk and wheat-reactive T cells in healthy individuals. Milk-reactive T cells were less frequent and expressed several markers of T cell exhaustion. In CD, but not UC patients, we identified higher frequencies of food-reactive T cells. These tended to be particularly increased in patients with disease manifestation in the upper gastrointestinal tract. In addition, we observed increased production of inflammatory cytokines of wheat-reactive T cells in CD, compared to healthy controls and UC patients. No ground-breaking differences were identified regarding the expression of exhaustion markers between IBD patients and controls.

Conclusion

Frequencies of food-reactive Th cells are increased in CD and show a proinflammatory cytokine pattern. The Th1-response of wheat-reactive T cells, characterized by high IFN-γ-secretion, might contribute to the perpetuation of chronic intestinal inflammation.
P064 IMU-838, a Small Molecule DHODH Inhibitor in Phase 2 Clinical Trial for Ulcerative Colitis, Shows Potent Anti-inflammatory Activity in Cell-Culture-Based and In Vivo Systems
Authors: Peelen E , Muehler A , Vitt D , Kohlhof H
Published: January 2022

Background

IMU-838 (vidofludimus calcium) is a well-tolerated and orally available small molecule inhibitor of dihydroorotate dehydrogenase (DHODH) and is currently in phase 2 clinical development for ulcerative colitis (UC). Here, we investigated the effect of IMU-838 on different immune cell subsets and analysed the mechanism of action in more detail in T cells.

Methods

The effect of IMU-838 on regulatory macrophages (Mregs) was investigated by adding IMU-838 or anti-TNFα antibodies (infliximab, adalimumab), alone or in combination, in a mixed lymphocyte reaction assay, determining Mregs (CD206+CD14+) by FACS analysis. To investigate the effect of IMU-838 on human T and B cells, PBMC were stimulated with PHA (48h) or Oligonucleotide 2006-PTO (70h), respectively.

An affinity-dependent effect of IMU-838 was assessed on CD8 T cells from OT-I and OT-III mice, containing a high and a low affinity T cell receptor (TCR), respectively, for OVA257-264 peptide. These cells were stimulated with ovalbumin (OVA) peptide loaded splenocytes or anti-CD3/anti-CD28 for 3 days. Oxidative phosphorylation (OXPHOS) and glycolysis in murine T cells was investigated by the Seahorse/Agilent technique in murine CD4 and CD8 T cells activated with anti-CD3/anti-CD28 with or without IMU-838.

Results

IMU-838 slightly induced Mregs, but strongly and dose-dependently decreased TNFα and IL-6 secretion. Anti-TNFα antibodies alone strongly induced Mregs, but also increased IL-6 levels. When IMU-838 was added, Mregs induction was even higher, and IL-6 was reduced. Besides reducing proinflammatory cytokines secreted by macrophages, IMU-838 also inhibits proliferation of activated B and T cells, as well as inflammatory T cell cytokine secretion, IL-17A, IL-17F and IFNγ, and increased apoptosis up to 3-fold compared to vehicle control. By investigating IMU-838’s role on affinity related activity status of T cells, it was shown that IMU-838 strongly inhibited cell proliferation in the peptide stimulated high affinity, but not in the low affinity, CD8 T cells. Peptide stimulated high affinity T cells show an upregulation of OXPHOS and glycolysis pathways, which are involved in the generation of the cell’s energy supply, compared to low affinity cells. IMU-838 strongly inhibited OXPHOS and glycolysis in both CD4 and CD8 T cells.

Conclusion

IMU-838 reduces the proinflammatory immune cell response by inducing Mregs, reducing pro-inflammatory cytokine secretion and reducing immune cell proliferation. DHODH is only important in cells that received a strong stimulus and are highly metabolically active. Therefore, IMU-838 will only specifically target these cells and will still allow for a normal immune response, representing a huge safety advantage for treatment of IBD patients.
P071 Intestinal CD8+ T cell exhaustion is linked to disease activity in ulcerative colitis
Authors: Grigorian M , Salié H , Mayer L S , Globig A M , Hockenjos B , Krimmel L , Thimme R , Bengsch B , Hasselblatt P
Published: January 2022

Background

The immune pathogenesis of ulcerative colitis (UC) has been linked to imbalanced CD4+ T-cell responses while the contribution of CD8+ T cells is less well characterized. Chronically activated CD8+ T cells may become dysfunctional and acquire an exhausted phenotype during chronic viral infection or cancer. Recently, exhausted T cell (Tex) signatures have been proposed as a potential prognostic marker in autoimmune diseases. We therefore set out to characterize features of CD8+ T cell exhaustion in UC patients in more detail.

Methods

Lymphocytes were isolated from peripheral blood and sigmoid biopsies of UC patients in remission (n = 9) and active disease (n = 20). Cytokine production and expression of exhaustion-associated markers were analyzed ex vivo by flow cytometry. Results were correlated with disease activity (Mayo Score) and mucosal inflammation (assessed endoscopically and histologically [Nancy Score]). To further analyze the role of CD8+ T cells within the intestinal tissue microenvironment, Imaging Mass Cytometry (IMC) was applied on sigma biopsies (n= 21).

Results

Analysis of CD8+ Tex marker expression and functionality did not reveal major differences between peripheral blood lymphocytes isolated from UC patients with either active disease or remission as well as healthy controls. In contrast, in the sigmoid colon of patients with active UC, CD8+ T cell numbers were strongly increased. Moreover, expression of exhaustion markers was significantly increased in intestinal CD8+ T cells during active disease compared to remission. These cells also displayed dysfunctional cytokine profile patterns, as assessed by an established functional exhaustion score. This phenotype also closely correlated with clinical, endoscopic and histological markers of UC disease activity. The findings were confirmed by IMC, which revealed the presence of several clusters of CD8+ T cells with an exhaustion signature in inflamed colonic tissues.

Conclusion

Our findings indicate that inflammation in the intestines of UC patients is associated with the induction of CD8+ T-cell responses that phenotypically and functionally resemble Tex. The functional relevance, prognostic value and therapeutic relevance of these exhausted CD8+ T cell signatures in UC need to be analyzed in future studies.
P077 Different granulocyte subsets are involved in the pathogenesis in Crohn’s disease fistula
Authors: Becker drs M , Koelink P , de Krijger M , Bemelman W , de Jonge W , Buskens C , Wildenberg M
Published: January 2022

Background

Early in inflammation granulocytes migrate from the circulation into inflamed areas and rapidly accumulate. They form an important role in first defense as well as the orchestration of downstream immune responses. In the circulation, several different subsets of granulocytes have been described, for example immature, intermediate and mature granulocytes. Surprisingly, despite their abundance and prominent function, this cell type is rarely studied from a tissue perspective, most likely due to their short lifespan ex vivo.

One example of gastrointestinal disease with an extremely high number of granulocytes is the occurrence of perianal fistula which contain a considerable amount of granulation tissue. As curettage of this tissue is part of the treatment, the fistula tract is good source to study tissue derived granulocyte subsets. The aim of this study was to characterize granulocyte subsets in the fistula tract.

Methods

Curettage material of perianal fistula tracts was obtained during surgical intervention from patients with CD (n=15) and cryptoglandular fistulas (n=5). From 2 CD patients venous blood was taken and a ficoll step was performed. Single-cell suspensions were stained with a 35-antibody panel, including CD66, CD14, CD33, CD16 and CD11b. Samples were analyzed using mass cytometry (CyTOF).

Results

Granulocytes, characterized as CD45+CD66a+ cells, were the main cellular component of the fistula tract (64%±24) compared to other immune cells. This was more pronounced in cryptoglandular fistula compared to CD fistula (81% vs. 67%; p=0.019). Within these cells we could differentiate between 4 subsets, mature granulocytes (CD33+CD11b+CD16+), intermediate (CD33intCD11b+CD16int), immature granulocytes (CD33intCD11b-CD16int) and eosinophils (CD33-CD11b-CD16-). Immature granulocytes were the most abundant in de fistula tract (31%±15), closely followed by the mature (30%±23) and intermediate (23±9). No significant difference was seen in the percentages of granulocyte subsets between CD and cryptoglandular fistula.

Conclusion

Here, we show the presence of different subtypes of granulocytes in fistula derived granulation tissue. We show a heterogeneous group of cells ranging from immature neutrophils to mature neutrophils. Due to their high number in active ulcerating tissue, further evaluation of granulocyte subsets and function could be an interesting target for medical treatment options.
P080 ADS024 attenuates weight loss and decreases disease activity in DSS-induced colitis
Authors: Chesnel L , Acton S
Published: January 2022

Background

Ulcerative colitis (UC) is a chronic, idiopathic inflammatory disease characterised by a dysregulated innate and adaptive immune response to gut microbiota that contributes to the perpetuation of colonic inflammation. ADS024 is an orally administered live biotherapeutic product (LBP), comprised of a single strain of Bacillius velezensis, being developed for potential application in the treatment of patients with UC.

Methods

Colitis was induced in 39 mice by exposure to 3% dextran sodium sulfate (DSS)-treated drinking water from Day 0 to Day 5; 6 control animals did not receive DSS. Animals were then dosed with vehicle (PBS; n=15) or ADS024 (n=12) twice daily for 14 days via oral gavage or with comparator anti-p40 mAb (n=12) once daily (IP) on Days 6, 9, 12, 15. All animals were monitored daily for body weight loss, diarrhea, blood in stool, and activity level. Each of these parameters was individually scored according to the disease activity index (DAI) scoring scheme. Animals underwent video endoscopy on Day 19 to assess colitis severity and stool consistency was scored during endoscopy. Following endoscopy on Day 19, all surviving animals were sacrificed, and blood was collected and processed for serum and analysed for metabolomic profile.

Results

Animals in which colitis was induced by exposure to 3% DSS demonstrated significant weight loss, most profoundly from Days 8 to 11, an increase in the composite DAI and increased mean colon weight:length ratios as compared to control animals. Animals treated with ADS024 had improved body weight, starting at 72 hours post-treatment and decreased DAI scoring over the course of the study compared to the vehicle control group (p<0.01 and p<0.05 respectively), comparable to treatment with anti-p40 mAb which has a mechanism of action similar to ustekinumab (Stelara®) approved for the treatment of UC. A trend towards a decrease in the mean colon weight:length ratio was observed in animals treated with ADS024, but there was no difference in mean endoscopy scoring compared to the vehicle control group. DSS treatment altered the serum biochemical profile at day 19 and ADS024 induced changes suggesting a reversion towards the control state.

Conclusion

Oral treatment with ADS024 improved body weight loss, reduced DAI, and resulted in a trend towards reduction in mean colon weight:length ratios in the DSS-induced mouse colitis model. In addition, ADS024 treatment led to a rebalancing of the systemic metabolomic shift caused by DSS exposure.
P087 Communicating Needs and Features of IBD Experiences (CONFIDE) Survey: Impact of Ulcerative Colitis Symptoms on Daily Life
Authors: Dubinsky M C , Schreiber S , Potts Bleakman A , Kayhan C , Ellis H , Hunter Gibble T , Flynn E , Suldecki G , Travis S
Published: January 2022

Background

Bowel urgency is the symptom described by patients as having the most impact on quality of life but is missing from most disease activity indices (Carpio et al., 2016). The Communicating Needs and Features of IBD Experiences (CONFIDE) study aims to increase understanding of patients’ experiences and the impact of IBD on their lives in the United States (US), Europe, and Japan. These data focus on US patients with moderately-to-severely active ulcerative colitis (UC).

Methods

An online, quantitative, cross-sectional survey was conducted with patients with moderately-to-severely active UC in the US in July 2021. Data included patient perspectives on their UC symptoms and the impact on their social life, work/school life, and ability to participate in sports/physical activities. Moderately-to-severely active UC was defined based on treatment, steroid use, and/or hospitalization history. Descriptive statistics (frequencies, percentages) summarise the data.

Results

200 (of 756 total contacted) patients (61.5% male, mean age 40.4, mean disease duration 7.9 years) completed the survey in the US, with 77% (n=153) of patients receiving advanced therapies (biologic or novel oral therapy). Patients declined participation in social events due to bowel urgency (43%), fear of urge incontinence (40%), increased stool frequency (29%), and blood in stool (15%) in the last three months (Fig. 1). Patients declined participation in sports/physical exercise due to bowel urgency (38%), fear of urge incontinence (36%), increased stool frequency (24%), and blood in stool (10%) in the last three months (Fig. 2). Patients declined participation in work/school due to bowel urgency (37%), fear of urge incontinence (42%), increased stool frequency (25%), and blood in stool (16%) in the last three months (Fig. 3). Out of 123 patients who have ever suffered from bowel urgency, in the last three months 42% stopped working for the day sooner than planned and 41% worked fewer hours. Notably, 76% of patients reported wearing a diaper/pad/protection at least once in the last three months due to fear of urge incontinence. Bowel urgency had no impact on work/school for 22% (n=27) of patients who had ever experienced bowel urgency (Fig. 4).

Conclusion

Bowel urgency and fear of urge incontinence are the most reported symptoms leading to patients with UC declining participation in work/school, social events, and sports/physical exercise. Over three quarters of the surveyed patients reported wearing diapers/pads/protection in the past three months due to fear of urge incontinence and only 22% of patients who had ever suffered from bowel urgency reported that it had no impact on work or school.
P096 Store-Operated Calcium Entry Controls Innate and Adaptive Immune Cell Function in Inflammatory Bowel Disease
Authors: Letizia M , Kaufmann U , Wang Y H , Gerbeth L , Sand A , Brunkhorst M , Böttcher C , Schlickeiser S , Fernández-Zapata C , Kunkel D , Siegmund B , Feske S , Weidinger C
Published: January 2022

Background

Inflammatory bowel disease (IBD) represents a major clinical challenge in need of new treatment modalities to improve patient care. Store-operated calcium entry (SOCE) is the predominant calcium influx pathway in T cells, regulating many of their functional properties. However, it is currently unknown whether the pharmacologic inhibition of SOCE is a suitable drug target in IBD. Therefore, we aimed to investigate the effects of SOCE inhibitors (SOCEi) on lymphocytes and epithelial cells isolated from IBD patients.

Methods

Peripheral blood mononuclear cells (PBMC), lamina propria lymphocytes (LPMCs) and epithelial cells were isolated from IBD patients undergoing colon resection. LPMCs or PBMCs were ex-vivo stimulated with Ionomycin/PMA ± SOCEi and subsequently stained with a panel of 37 immunological markers for mass cytometry acquisition. Ca2+ influx measurements were performed in order to assess the metabolic status of immune (PBMCs and LPMCs) and epithelial cells after SOCE blockade. Finally, murine models of colitis were used in order to investigate the effects of SOCE blockade on the induction of intestinal inflammation in-vivo.

Results

Data on B, T, NK, and myeloid cells revealed that treatment with SOCEi caused a dose-dependent inhibition of Ca2+ signaling in human gut-resident lymphocytes and PBMCs. The inhibition of SOCE attenuated the production of pathogenic cytokines including IL-2, IL-4, IL-6, IL-17, TNF-α and IFNγ by colonic T cells and ILCs, reduced the activation of B cells and the production of IL-6 by myeloid cells, without affecting the viability of primary human epithelial cells. Remarkably, mouse models of transfer colitis, in which SOCE signaling components have been blocked, revealed that SOCE is required for the induction of intestinal inflammation in-vivo.

Conclusion

Our data revealed for the first time that the cytokine production and the activation of several immune cell subtypes can be modulated by SOCE blockade in human intestinal inflammation, identifying SOCE as a novel therapeutic target in colitis.
P104 Icariin Attenuates Intestinal Inflammation through Inhibition of the Nuclear Factor-ĸB signaling
Authors: Soh H , Park S , Lee H J , Koh S J , Im J P , Kim J S
Published: January 2022

Background

Icariin is a natural flavonoid glucoside from plants in Epimedium family, known to have anti-oxidative and anti-inflammatory activities. This study investigated the effects of Icariin on nuclear factor-kappa B (NF-ĸB) signaling pathway in intestinal epithelial cells (IEC) and macrophages, and in acute and chronic murine colitis models.

Methods

Human IEC line COLO 205 and murine macrophage cell line RAW 264.7 were pretreated with Icariin and then stimulated with tumor necrosis factor-α (TNF-α) and lipopolysaccharide respectively for in vitro studies. The expression of pro- and anti-inflammatory cytokines were evaluated by real-time RT-PCR (TNF- α, IL-6, IL-8, IL-12, and IL-10), and the effect of Icariin on NF-ĸB signaling pathway was examined by immunoblot analysis to detect IĸBα phosphorylation and degradation. Dextran sulfate sodium (DSS)-induced acute colitis in C57BL/6 wild-type mice and chronic colitis in IL-10-/- mice were treated with or without Icariin for in vivo studies. The severity of colitis was evaluated by daily body weight change, colon length, and colonic tissue histopathology.

Results

Icariin significantly attenuated the production of pro-inflammatory cytokines and degeneration of anti-inflammatory cytokines, and alleviated IĸBα phosphorylation and degradation in both IEC and murine macrophages stimulated by TNF-α and LPS, respectively. Icariin also significantly reduced the severity of colitis in both DSS-induced acute colitis model and chronic colitis in IL-10-/- mice models.

Conclusion

Icariin inhibits NF-ĸB signaling in both IECs and macrophages, and attenuates experimental murine colitis. These results suggest that Icariin may be a potential novel therapeutic agent for inflammatory bowel disease.
P141 Factors Associated with Time to Clinical Remission in Pediatric Luminal Crohn’s disease: a Retrospective Cohort Study
Authors: Sassine S , Fadela Zekhnine S , Qaddouri M , Djani L , Cambron-Asselin C , Savoie-Robichaud M , Lin Y F , Grzywacz K , Groleau V , Dirks M , Drouin É , Halac U , Marchand V , Girard C , Courbette O , Patey N , Dal Soglio D , Deslandres C , Jantchou P
Published: January 2022

Background

The natural evolution of Crohn’s disease is incompletely understood in the pediatric population. Data on factors influencing time-to-remission are very limited in the literature. The aim of this retrospective cohort study was to describe the time to clinical remission in children with Crohn’s disease as well as changes over the past decade and to identify factors associated with time to clinical remission.

Methods

Patients under 18 years old diagnosed between 2009 and 2019 were included. All data were collected from the patients’ medical records and the CHU Sainte-Justine inflammatory bowel disease registry. Survival analyses and linear regression models were used to assess the impact of clinical, laboratory, endoscopic, histological and therapeutic factors on time to clinical remission.

Results

654 patients were included in the study. There was no change in the time to clinical remission over the past decade. Female sex in adolescents (ajusted bêta regression coefficient (aβ)= 31.8 days, p= 0.02), upper digestive tract involvement (aβ= 46.4 days, p= 0.04), perianal disease (aβ= 32.2 days, p= 0.04), presence of active inflammation on biopsies (aβ= 46.7 days, p= 0.01) and oral 5-ASA exposure (aβ=56.6 days, p= 0.002) were all associated with longer time to clinical remission. However, antibiotic exposure (aβ= -29.3 days, p=0.04), increased eosinophils on biopsies (aβ= -29.6 days, p=0.008) and combination of exclusive enteral nutrition and TNF- alpha inhibitors as induction therapy (aβ= -36.8, p=0.04) were associated with shorter time to clinical remission.

Figure 1: Kaplan-Meier curve representing time to clinical remission in patients 13 years old and older by sex.

Figure 2: Kaplan-Meier curve representing the time to clinical remission of patients according to the first induction treatment administered.

Conclusion

Time to clinical remission did not improve during the decade and was associated with baseline clinical and histological data and treatment strategies. Combination of enteral nutrition and TNF-alpha inhibitors was associated with faster clinical remission.
P154 Evaluation of serum protein FingerPrint biomarkers of collagen, citrullinated vimentin and calprotectin in patients with inflammatory bowel disease
Authors: Luo Y , Saini J , Mortensen J , Bay-Jensen A , Karsdal M , Greenawalt D , Harris S
Published: January 2022

Background

Inflammatory bowel disease (IBD) is characterized by chronic inflammation leading to extracellular matrix remodeling, reactive stroma and activated inflammatory cells. In the pursuit of non-invasive biomarkers of disease severity and treatment response for IBD, we evaluated Protein FingerPrint Assay (PFA) of collagen formation and degradation (PRO-C3, C3M, PRO-C4, C4M), neutrophil elastase cleaved calprotectin (CPa9-HNE) and citrullinated vimentin degradation (VICM) in serum collected from 2 clinical trials in Ulcerative Colitis (UC) and Crohns Disease (CD) (NCT01466374 and NCT01294410) and matched healthy volunteers (NHV).

Methods

PFA were generated from serum of 88 UC, 120 CD at baseline and at Day 78 of treatment and 78 NHV. Clinical remission in UC was defined as Mayo score 2 with no individual subscore >1. Clinical response as reduction from baseline in Mayo score 3 and 30%, and decrease from baseline in rectal bleeding subscore 1 or absolute rectal bleeding subscore 1. Clinical remission in CD was defined by an absolute CD Activity Index [CDAI] score < 150 and clinical response as a reduction in CDAI100 points from baseline or an absolute CDAI score < 150. Endoscopic remission was defined as SES-CD of 0. Statistical analysis was performed on all markers between diseased and NHV, pooled responders/remitters vs non-response independent of treatment arm and longitudinally.

Results

At baseline, C3M, C4M and PRO-C4 had significantly higher median levels in UC and CD patients than NHVs. CPa9-HNE and VICM were significantly higher in CD compared to NHV, an increase in these markers was observed but did not meet significance in UC patients (Figure 1). Decrease of VICM from baseline at Day 78 were associated with remission (median -55%, p<0.05) in UC, while VICM decrease from baseline (median -56%, p<0.05) at Day 78 was associated with CD patients achieving endoscopic remission. In CD C3M and C4M was present at lower levels at baseline and during study in patients with clinical remission compared with those without remission. SES-CD scores were positively correlated with C4M, C3M, CPa9-HNE and PRO-C4 at both baseline and Day 78. CD patients achieving endoscopic remission had lower levels of C3M, C4M, PRO-C4 and CPa9-HNE at both baseline and Day 78.

Conclusion

C4M, C3M, PRO-C4, CPa9-HNE and VICM were found at higher levels in serum of IBD patients. VICM demonstrated potential as a biomarker for monitoring disease activity changes. Association of C4M, C3M and PRO-C4 with endoscopic remission in CD patients suggest that these markers may predict disease progress or regression in CD patients. However, further studies are warranted to evaluate these biomarkers in IBD patients.
P164 Assessment of awareness and compliance with the recommendations of primary and secondary prevention of cancer in patients with Inflammatory Bowel Disease
Authors: Tulewicz-Marti E , Łodyga M , Stępień-Wrochna B , Lewandowski K , Rydzewska G
Published: January 2022

Background

Patients with Inflammatory Bowel Disease (IBD), including Crohn’s disease (CD) and ulcerative colitis (UC), are at risk of developing malignancies, so prevention and adherence to cancer screening may improve its detection. The aim of this study was to asses the compliance with medical recommendations, especially primary and secondary prevention of cancer.

Methods

This prospective, one-center study was carried out between June and October 2021r. amongst patients from Department of Internal Medicine and Gastroenterology with IBD Division, Central Clinical Hospital of Ministry of Interior Affairs and Administrations, Warsaw, Poland. Patients were asked to complete anonymous questionnaire which included 42 questions concerning lifestyle, cancer risk factors, cancer history, and checkups in patients with IBD.

Results

A total of 190 patients with IBD were included in survey. 117 (61.6%) had CD, 67 (35.3%) UC and 6 (3.2%) unclassified disease. 43.7% were women and more than a half of them had >8 years of the length of the disease. In total 84.2% were taking mesalazine, 48.4% immunosupresive medication and 77.9% biological treatment. Statistically significantly (p=0.034) more patients with CD than those with UC practiced regular physical activity (respectively 65.8% and 48.5%). Patients with UC more often were non-smokers (76.7% vs 92.5% for CD and UC respectively; p=0.007). 39.7% of patients with CD and 27.3% with UC declared alcohol consumption (p=0.128). Only 6% of patients responded not to use sun protection measures when sun-bathing but 24.8% with CD and 34.4% with UC didn’t have regular full-body skin examinations. 78.7% of patients had abdominal ultrasound scan performed in the last year. Out of 52.7% who had colonoscopy recommended, 49.3% of patients declared that the examination was carried out (36.8% of CD and 65.5% of UC; p= 0.038) and another endoscopy was scheduled (18.6% of CD and 48.4% of UC; p<0.001). According to respondents cholangio-MRI was recommended to 14.3% patients with CD and 6.2% with UC (p=0.219). Only 1 patient in both groups did not perform this examination. Most of the examinations were recommended by gastroenterologists. Moreover, in the last 12 months 59.6% of female answerer declared to have ultrasound imagining of the breast and 98.7% gynecological examination performed (65.4% had regular Pap-test screening). 82.3% of respondents knew about HPV vaccination but only 25% were vaccinated.

Conclusion

Our study reveals that many of IBD patients are exposed on risk factors of malignancies. Screening for cancer and regular checkups should be regularly performed. Primary prevention, such as HPV vaccinations, should be reminded to all patients.
P174 Clostridium innocuum Infection in Inpatients with Inflammatory Bowel Disease
Authors: Le P H , Chiu C H , Chiu C T
Published: January 2022

Background

Clostridium innocuum (CI) infection can lead to creeping fat in Crohn’s disease and is associated with intestinal strictures. At present, no clinical study ever has evaluated the role of CI infection in inflammatory bowel disease (IBD).

Methods

In this retrospective cohort study, we enrolled hospitalized IBD patients with culture results for both CI and Clostridioides difficile (CD) in a medical center between October 2019 and April 2021. They were divided into the CI (CI+/CD−), control (CI−/CD−), coinfection (CI+/CD+), and CD (CI−/CD+) groups. We analyzed the risk factors, clinical presentations, and outcomes by comparing the CI and control groups.

Results

We enrolled a total of 90 patients, including 22, 39, 13, and 16 patients in the CI, control, coinfection, and CD groups. The incidence rates of CI (CI+) and CD (CD+) were 39% (35/90) and 32% (29/90), respectively. We analyzed the differences between CI and control groups. We identified the use of steroid (77.3% vs. 46.2%, P = 0.018) and 5-aminosalicylic acid (90.9% vs. 64.1%, P = 0.022) as risk factors of CI infection. Clinical analysis showed that more patients in CI group presented with bloody stool (77.3% vs. 51.3%, P = 0.046). Although CI group had significantly lower overall occurrence of intraabdominal abscess (0% vs. 17.9%, P = 0.042), it showed a lower clinical remission rate (50% vs. 87.5%, P = 0.044) and higher Mayo score at the end of follow-up (10 points vs. 3 points, P = 0.008) in ulcerative colitis.

Conclusion

CI infection may lead to a poorer clinical remission in ulcerative colitis. We should take it into consideration in IBD patents with active inflamamtion or refractory diarrhea with or without CD infection. Precise identification of CI is imperative to guide approproate antimicrobial therapy because of its intrinsic vancomycin resistance nature.
P185 The role of neutrophil-lymphocyte ratio, neutrophil-monocyte ratio, and lymphocyte-monocyte ratio in estimating disease activity in patients with ulcerative colitis
Authors: Sina Dr M , Pemaj X , Akshija I , Prifti S
Published: January 2022

Background

Determining disease activity in ulcerative colitis (UC) patients is important in order to reduce surgery rate and mortality. Colonoscopy has the disadvantage of being invasive, time consuming and expensive. Several laboratory markers have been proposed in order to evaluate UC disease activity.

Methods

The aim is to investigate the utility of neutrophil-lymphocyte ratio (NLR), neutrophil-monocyte ratio (NMR), and lymphocyte-monocyte ratio (LMR) in differentiating active UC from UC in remission. This is a prospective study carried out at a hospital center in Albania, from 2019–2020. Demographic and endoscopic data along with complete blood counts were collected for each UC patient. Based on Mayo score system, patients were classified as active UC (total Mayo score >2 points) and UC in remission (total Mayo score ≤ 2 points). Pearson correlation coefficient was used to analyze associations between parameters. The sensitivity, specificity, and cut-off values were assessed using a receiver operating characteristic curve analysis.

Results

65 UC patients, with a mean age of 45.05 ± 15.5 (16–85) years, of whom 52.3% were females and 47.7% males, were enrolled in this study. 21.4% were in remission and 63.4% were classified as active UC disease. We found a positive correlation between NLR, NMR and disease activity [(r=0.295, p=0.002); (r=0.241, p=0.011) respectively]. NLR and NMR were also significantly higher in active disease group compared to UC in remission (p=0.016 and p=0.03 respectively). ROC analyses revealed that NLR (AUC=0.664, 95% CI, 0.557–0.772) and NMR (AUC=0.644, 95% CI, 0.531–0.756) are satisfactory in differentiating active UC from UC remission patients. A NLR cutoff value of 2.1 had a sensitivity of 60% and specificity of 61% for active UC, while a NMR cutoff value of 12.2 had a sensitivity of 63% and a specificity of 61% for active UC. We didn’t found any correlation between LMR and disease activity

Conclusion

This study shows that NLR and NMR are significantly associated with active UC disease. Given the fact that complete blood count is widely available and inexpensive, NLR and NMR may play a role in differentiating active UC from UC in remission.