bluuebunny/crossref_metadata_2025 huggingface dataset:
P018 Per- and polyfluoroalkyl substances (PFAS) are significantly increased in patients with late-onset of ulcerative colitis
Authors: Fart F , Salihovic S , McGlinchey A , Orešič M , Halfvarson J , Hyötyläinen T , Schoultz I
Published: January 2020

Background

Environmental factors have been implicated in the pathogenesis of inflammatory bowel disease (IBD), particularly late onset disease. Per- and polyfluoroalkyl substances (PFAS) are man-made chemicals with a long biological half-life that have been extensively used since the 1950s and have been proposed to interfere with the bile acid synthesis. Therefore, to investigate if late onset IBD correlates with higher PFAS levels, we measured serum levels of PFAS and bile acids in patients diagnosed with IBD later in life.

Methods

Serum samples were collected from patients diagnosed with ulcerative colitis (n = 20) and Crohn’s disease (n = 20) at the age of ≥55 years. Blood donors (n = 20) were used as healthy controls and were matched by gender and age. The levels of PFAS and bile acids were assessed by ultra performance liquid chromatography coupled to a triple quadrupole mass spectrometer.

Results

The total amount of PFAS was significantly higher in patients with ulcerative colitis compared with healthy controls (p = 0.021) or patients with Crohn’s disease (p = 0.015). No difference was found in total PFAS levels between Crohn’s disease patients and healthy controls (p = 0.841). Seven out of 30 bile acids correlated to the total PFAS level.

Conclusion

Our results demonstrate that PFAS levels are increased in patients with late-onset of ulcerative colitis compared with Crohn’s disease patients and healthy controls. This finding indicates that PFAS might represent an environmental risk factor for ulcerative colitis. However, additional studies assessing the functional consequences of increased PFAS in late-onset ulcerative colitis are required to confirm this hypothesis.
P024 Evaluation of the clinical course of patients diagnosed with primary sclerosing cholangitis: single centre study
Authors: Wiecek S , Wojtyniak A , Pindur B , Grzybowska-Chlebowczyk U
Published: January 2020

Background

Primary sclerosing cholangitis (PSC) is a rare liver cholestatic disease with unknown aetiology, severe clinical course and poor prognosis. PSC most often co-occurs with inflammatory bowel diseases (IBD), especially ulcerative colitis (UC). The aim of the study. We aimed to analyse the clinical course of patients diagnosed with PSC hospitalised in the Department of Gastroenterology, Medical University of Silesia in Katowice in 2009–2018.

Methods

The analysis included 30 patients, aged from 7 to 18 years (mean age 13.5 years), 21/30 boys (70%) and 9/30 girls (30%), diagnosed with PSC. The analysis included age of diagnosis, clinical symptoms, course of the disease, laboratory and imaging results and complications.

Results

The average diagnosis age was 13 years. 22/30(73.3%) patients suffered from UC and 4/30 (13.3%) were diagnosed with Crohn’s disease. 2/30 (6.66%) patients remained IBD-free. Among them 7/30(23.3%) had an overlap syndrome—primary sclerosing cholangitis and autoimmune hepatitis. At the time diagnosis 6/30 (20%) patients presented abdominal pain, which was the most common symptom, while 16/30 (57%) were asymptomatic but had abnormal laboratory test results. When PSC was detected before IBD, patients had complications more often compared with those diagnosed with IBD first or PSC and IBD at the same time. Moreover, complications were observed more often in patients with changes characteristic of PSC visible in their first USG than in patients without mentioned abnormalities in first USG.

Conclusion

Patients diagnosed with PSC before IBD diagnosis are more likely to have a more aggressive course of the disease. Monitoring liver markers in IBD patients is important since most PSC cases are asymptomatic and their elevation might be the first sign of the disease.
P032 Increased endoplasmic reticulum stress-specific chaperones characterise CD fibrosis epithelium tissues and participate to in vitro induction of intestinal fibrobast differentiation
Authors: Vieujean S , Hu S , Bequet E , Salee C , Massot C , Bletard N , Pierre N , Quesada calvo F , Baiwir D , Mazzucchelli G , De Pauw E , Delvenne P , Meuwis M A , Louis E
Published: January 2020

Background

Intestinal fibrosis is a complication of Crohn’s disease (CD) characterised by myofibroblasts and extracellular matrix accumulation within the submucosa and smooth muscles, leading to bowel strictures. No medical treatment exists to treat or reverse intestinal fibrosis leading often to surgical resection. The potential role of intestinal epithelium in the fibrotic process remains poorly defined.

Methods

We performed a pilot study on ileal fibrostricturing CD surgical samples (n = 5), comparing the proteome of surface epithelium isolated by laser capture microdissection in normal and fibrotic zones. Confirmation of the specific protein increases was obtained by immunohistochemistry in colonic and ileal samples of CD (n = 44) compared with healthy subjects (n = 40), as well as in intestinal epithelial cell line under induced endoplasmic reticulum (ER) stress. A model of fibroblast to myofibroblast differentiation induction was also challenged using preconditioned media of intestinal epithelial cells after a pulsed ER stress.

Results

Label-free proteomics revealed high ER stress in the epithelium surrounding fibrotic bowel wall, involving anterior gradient protein 2 homolog (AGR2) and 78 kDA glucose-regulated protein (BiP). Confirmation of both proteins increase was obtained by immunohistochemistry. ER stress induction in intestinal epithelial cells was associated with an intracellular increase of AGR2, BiP and ER stress markers as sXPB1 and CHOP. AGR2 was also detected in the culture medium of these epithelial cells and myofibroblast differentiation was obtained using this culture medium.

Conclusion

The increase of ER stress proteins observed in fibrostenosing tissues together with these preliminary evidences of fibroblast to myofibrobast differentiation obtained by paracrine action of intestinal epithelial cell preconditioned to ER stress induction suggest a role of epithelial ER stress in Crohn’s disease intestinal fibrosis.
P033 Combination of vedolizumab with tacrolimus is more efficient in reducing intestinal inflammation than vedolizumab alone in the DSS-induced acute colitis mouse model
Authors: Manzini R , Atrott K , Schwarzfischer M , Laimbacher A , Lang S , Rogler G , Scharl M , Spalinger M R
Published: January 2020

Background

The humanised monoclonal antibody vedolizumab is used in the treatment of ulcerative colitis (UC) and Crohn’s disease (CD). Vedolizumab reduces intestinal inflammation through inhibition of the integrin heterodimer α 4β 7, responsible for the homing of T cells to the intestinal mucosa. Recent studies have also shown a possible involvement of vedolizumab in the regulation of the innate immune system. Particularly in CD, only a fraction of patients respond to vedolizumab treatment, and combination therapy with immunosuppressant drugs, such as the calcineurin-inhibitor tacrolimus, might prove beneficial. The aim of this study was to assess if co-treatment of vedolizumab and tacrolimus is more efficient in reducing intestinal inflammation in an acute colitis mouse model and to unravel the underlying molecular mechanisms.

Methods

NOD-SCID-SGM3 mice were reconstituted with human CD34+ cells and treated with 3% dextrane sodium sulphate (DSS) in drinking water to induce acute colitis. Mice were treated with vedolizumab alone (30mg/kg, inject 3 days prior to DSS-start and 50mg/kg at day 0 and at day 4 of DSS-treatment), tacrolimus alone (1mg/kg/day intraperitoneally), or a combination of tacrolimus and vedolizumab during colitis induction.

Results

As expected, DSS-treatment induced colitis in mice as observed by weight loss, diarrhoea, colon shortening, and endoscopic signs of inflammation categorised by the MEICS score. This translated histologically to an increased immune cell infiltration and epithelial erosion. Vedolizumab and tacrolimus treatment alone did not significantly reduce colitis severity, although endoscopy showed slightly less severe inflammation in vedolizumab-treated mice. Combination of vedolizumab and tacrolimus, however, clearly reduced colonoscopy and histology scores. DSS-treatment increased the number of CD3 T cells and CD68 macrophages in the intestine, an effect counteracted by vedolizumab or tacrolimus alone and further pronounced by combination treatment. Particularly vedolizumab treatment, either alone or in combination, caused clear reduction of pro-inflammatory M1 macrophages. Additionally, vedolizumab alone or combined decreased the levels of intestinal epithelial cell apoptosis as indicated by staining for cleaved caspase-3.

Conclusion

Our data demonstrate that the anti-inflammatory effect of vedolizumab is potentiated by co-treatment with tacrolimus. Notably, the combination of both drugs was more efficient in reducing T-cell and macrophage infiltration into the intestine. This indicates that the combination of vedolizumab with immunosuppressant drugs might prove beneficial for patients that do not respond to vedolizumab-only therapy.
P037 Antagonism of chemokine receptor CCR9 synergises with anti-TNFα immunotherapy to reduce inflammation in the MDR1a−/− mouse model of colitis
Authors: Campbell J , Ertl L , Ebsworth K , McMahon J , Zhang P , Chhina V , Charo I , Schall T
Published: January 2020

Background

The development of anti-TNFα immunotherapies has led to great advances in quality of life for Crohn’s and colitis patients and has allowed a reduction in steroid use for these diseases. However, a significant proportion of IBD patients is resistant to anti-TNFα agents, or becomes resistant to such treatments over time. Preclinical research and clinical trials have suggested that CCR9 antagonism can effectively reduce colon inflammation. CCR9 is expressed on a subset of T cells dedicated to monitoring the immune status of the intestine. When activated by its only known chemokine ligand, CCL25/TECK, CCR9 triggers the adhesion of gut-specific T cells to vascular endothelium and the subsequent ingress of these cells to intestinal tissues. Antagonism of this receptor inhibits the accumulation of T cells within inflamed intestinal epithelium and lamina propria. Since anti-TNFα and CCR9 antagonists act through entirely different mechanisms, we asked whether these two approaches might synergise and expand the effectiveness and scope of each therapy.

Methods

We used a piroxicam-accelerated version of the MDR1a−/− spontaneous colitis model in mice. In this model, 7week MDR1a−/− mice (or WT FVB controls) receive piroxicam in powdered chow for 10 days. Mice are monitored both during and after the piroxicam feeding period, for a total of 21 days. Treatment groups (eight mice each) include an antibody (a saturating dose of either anti-TNFα or its isotype-matched control) and a CCR9 antagonist (or its vehicle alone as a control). Body weight and diarrhoea score were recorded throughout the entire 21 days period, after which colons were obtained from sacrificed animals for weighing, length measurement and imaging.

Results

We have found that combined dosing of piroxicam-fed MDR1a−/− mice with a CCR9 antagonist and an anti-TNFα MAb significantly reverses loss of body weight, diarrhoea score and colon inflammation. The combined dosing was more effective than anti-TNFα or CCR9 antagonist alone, as measured by colon length/weight ratio, neutrophil accumulation in the colonic epithelium and reversal of the loss in body weight characteristic of this model.

Conclusion

These data suggest that combining a CCR9 antagonist with existing anti-TNFα treatment regimens may extend the benefits of anti-TNFα immunotherapy to a larger class of patients suffering from IBD and may prolong the effectiveness of anti-TNFα immunotherapy for those patients already undergoing such treatments.
P041 Melatonin controls microbiota in colitis through Toll-like receptor 4 signalling
Authors: Park Y S , KIm S W , Cheon J H
Published: January 2020

Background

Microbial dysbiosis has long been postulated to be associated with the pathogenesis of inflammatory bowel disease (IBD). Although evidence supporting the anti-colitic effects of melatonin have been accumulating, it is not clear how melatonin affects the microbiota. Herein, we investigated the effects of melatonin on the microbiome in colitis and identified involvement of Toll-like receptor (TLR) 4 signalling in the effects.

Methods

Colitis was induced in 8-week-old wild-type (WT) and TLR4 KO mice by administration of 2.5% dextran sulphate sodium (DSS) in drinking water and intraperitoneal (i.p.) treatment with melatonin (Mel) or vehicle (Veh). Body weight changes were monitored daily.

Results

DSS-treatment for 10 days significantly increased body weight loss in the vehicle-treated group of both wild-type and TLR4 KO mice. Melatonin improved dextran sulphate sodium (DSS)-induced colitis and reverted microbial dysbiosis in wild-type (WT) mice but not in TLR4 knockout (KO) mice. Induction of goblet cells was observed with melatonin administration, which was accompanied by suppression of Il1b and Il17a and induction of melatonin receptor and Reg3β, an antimicrobial peptide (AMP) against Gram-negative bacteria. In vitro, melatonin treatment of HT-29 intestinal epithelial cells promotes mucin and wound healing and inhibits growth of Escherichia coli.

Conclusion

we showed that melatonin significantly increases goblet cells, Reg3β and the ratio of Firmicutes to Bacteriodetes by suppressing Gram-negative bacteria through TLR4 signalling. Our study suggests that sensing of bacteria through TLR4 and regulation of bacteria through altered goblet cells and AMPs is involved in the anti-colitic effects of melatonin. Melatonin may have use in therapeutics for IBD.
P047 Differences in extra cellular matrix remodelling in highly active Crohn’s disease and ulcerative colitis
Authors: Domislović V , Mortensen J H , Karsdal M A , Brinar M , Manon-Jensen T , Krznarić Ž
Published: January 2020

Background

Ulcerative colitis (UC) and Crohn’s disease (CD) require continuous evaluation of disease activity and response to therapy. Current gold standard is endoscopy; therefore it is important to investigate biomarkers associated with endoscopic disease activity. Extra cellular matrix (ECM) consists of basement membrane (BM) and interstitial matrix (IM). BM consists mainly of type IV collagen, while IM of types I, III and V collagen. Pathological environment leads to impaired remodelling, structure, quality and function of the ECM. We investigated biomarkers of collagen degradation and formation and their association with endoscopic disease activity and in patients with CD and UC.

Methods

In this cross-sectional study, we measured six biomarkers of ECM remodelling in 94 IBD patients (60 with CD and 34 UC). Biomarkers of type III collagen degradation (C3M) and formation (PRO-C3), type IV collagen degradation (C4M), formation (PRO-C4), type V collagen formation (PRO-C5) and PROC23 were measured in serum by ELISA. Inflammatory activity was measured using endoscopic scores; SES-CD for CD, MES and modified MES (mMES) for UC. Patients were divided in remission and mild activity group vs. moderate-to-severe activity (SES-CD < 3 vs. ≥3 and MES < 2 vs. ≥2) Student t-test and correlation analysis were applied in statistical analysis.

Results

CD patients with moderate and severe disease had lower levels of PRO-C3 (14.7 ± 10.5 vs. 9.9 ± 4.6, p = 0.04), and higher levels of C4M (26.54 ± 10.5 vs. 37.8 ± 20, p = 0.009) compared with patients in remission and mild disease. Biomarker of type III collagen turnover (C3M/PROC3) showed higher levels in moderate and severe active disease in CD (1 ± 0.5 vs. 1.8 ± 1.1, p = 0.006). UC patients with moderate to severe activity showed higher levels of type III collagen turnover biomarker (C3M/PROC3) (1.1 ± 0.4 vs. 1.97 ± 1, p = 0.049). C4M correlated positively (r = 0.28, p = 0.03) and PRO-C3 correlated negatively (r = −0.29, p = 0.03) to SES-CD and combined in a multivariate regression model (r = 0.39, p = 0.009). C3M (r = 0.60, p = 0.004) and C4M (r = 0.36, p = 0.039) correlated positively with mMES.

Conclusion

In highly active CD, there is increased type IV collagen degradation and reduced type III collagen formation. Both biomarkers significantly correlate with SES-CD, with increased correlation when combined together. Type III collagen turnover is increased in highly active CD and UC. Biomarkers of types III and IV collagen degradation correlate significantly with mMES. Biomarkers of types III and IV collagen could be used to differentiate highly active CD from UC. Our findings suggest that biomarkers of basement membrane and interstitial matrix remodelling may be used as surrogate markers for monitoring of disease activity for UC and CD.
P050 Analysis of 61 exclusive enteral nutrition formulas used for induction of remission in Crohn’s disease: new insights on dietary disease triggers
Authors: Logan M , Gkikas K , Svolos V , Nichols B , Milling S , Ijaz U , Hansen R , Russell R K , Gerasimidis K
Published: January 2020

Background

Exclusive enteral nutrition (EEN) is an effective treatment of Crohn’s disease (CD). We hypothesise that food ingredients which are included in EEN formulas are less likely to initiate a disease flare and that their dietary elimination is not essential for disease amelioration.

Methods

We performed a compositional analysis of EEN formulas with evidence of clinical efficacy for the induction of remission in active CD. Macronutrient content was compared against the dietary reference values (DRVs), the UK National Diet and Nutrition Survey (NDNS) and children with CD. Food additives included with EEN formulas were cross-referenced against the FAO/WHO database.

Results

Sixty-one formulas were identified with variable composition [carbohydrates (23–89%), protein (8–30%), fat (0–50%)], Figure 1. Modified starches (e.g. maltodextrin), milk protein and vegetable/plant oils were the commonest sources of carbohydrate, protein and fat, respectively, Figure 2. The n-6:n-3 fatty acid ratio varied from 0.3 to 46.5. Fifty-six unique food additives were identified contained within EEN formulas (median per formula: 11), Figure 3. All formulas were lactose, gluten free. Eighty per cent lacked fibre. The five commonest food additive functional classes were emulsifiers, stabilisers, antioxidants, acidity regulators, and thickeners. Food additives, implicated in CD aetiology, were present in formulas [modified starches (100%), carrageenan (23%), carboxymethyl cellulose (14%) and polysorbate 80 (5%)]. EEN formulas derived 7.9% less energy from saturated fat than participants from the NDNS (p = 0.001). CD children consumed more sugars, total/saturated fat than the EEN content.

Conclusion

We provide a list of food ingredients which are unlikely, in the amount provided within EEN, to trigger CD activity. Current perceptions about the role of these ingredients in CD management are challenged.
P058 The lived experience of family planning of female patients diagnosed with inflammatory bowel disease and their partners during key reproductive stages: a qualitative study
Authors: Czuber-Dochan W , Homer R , Brookes M , Selinger C , Purewal S , Chapman S , Steed H
Published: January 2020

Background

Inflammatory bowel disease (IBD) is a chronic illness affecting patients in their childbearing years. The physical effects of IBD on fertility and pregnancy in IBD in remission (e.g. disease is well controlled during conception and throughout pregnancy) are similar to the normal population. However, many women with IBD have high pregnancy-related anxieties and are more likely not to have children compared with women without IBD. The reasons cited for not having children include high levels of pregnancy-related anxieties have been insufficiently explored. The study aimed to explore the lived experience of family planning of women with inflammatory bowel disease (IBD) and their partners with or without IBD, during the reproductive stages of pre-conception, pregnancy and the postnatal period.

Methods

Descriptive phenomenology was used to conduct face-to-face in-depth individual interviews. Purposive sampling was used to select participants with a maximum variation of different demographic and clinical factors, e.g. age, sex, UC/CD diagnosis, disease duration, surgery and geographic location. The NVivo 12 software programme was used to manage the data and Colaizzi’s framework was utilised in thematic data analysis.

Results

Twenty-four participants (21 women 11CD/10UC and three partners) were recruited from out-patient clinics (22 participants) or through the Crohn’s and Colitis UK website (two participants). Women, average age 31 years old (range 27–38), were at different family planning stages: pre-conception six women (three actively planning family and three voluntarily childless); pregnant eight women and two partners; and postpartum seven women and one partner. Three women’s partners, age 32–39, were recruited allowing for additional perspective of the experience being captured. In total, 19 h of interviews data were collected. Six themes were identified: (1) being diagnosed and controlling IBD symptoms, (2) relationship and family planning, (3) sources of information,

(4) worries and concerns about pregnancy, (5) post pregnancy care and problems and (6) ways of improving care. Women in pre-pregnancy stage and pregnant expressed a need for more information around these themes: (1) their medication and the impact of IBD on the baby, (2) the genetic risk of passing the disease on. While pregnant and during post-partum stage, women identified a greater need for practical advice and support in relation to breastfeeding and looking after the baby.

Conclusion

Information specific to family planning stages need to be provided, to help women and their partners make the informed decision about family planning. Those who decided not to go down the family route also expressed a need for counselling and support.
P067 Proteins citrullination and Crohn’s disease: PAD4 but not PAD2 is a strong marker of ileal inflammation
Authors: Dragoni G , Creyns B , De Hertogh G , Verstockt B , Wollants W J , Ke B J , Marcellis L , Matteoli G , D’Hoore A , Galli A , Ferrante M , Vermeire S
Published: January 2020

Background

Citrullination is a post-translational modification of proteins, mediated by enzymes called PAD (peptidylarginine deiminases). The immune system can attack citrullinated proteins, leading to autoimmune diseases such as rheumatoid arthritis, multiple sclerosis and ulcerative colitis, and the activity of PAD2 and PAD4 in innate immune cells has been demonstrated for these disorders. Recently, high levels of PAD2 have been described in activated fibroblasts in the context of liver fibrosis. We therefore investigated the role of PAD2 and PAD4, both in inflammatory and fibrotic contexts of ileal Crohn’s disease (CD).

Methods

We obtained ileal transmural samples from patients operated for stricturing ileal CD. Three different macroscopic areas within each resection specimen (i.e. proximal normal ileum, inflamed ileum and fibrotic ileum) were selected and histologically confirmed by an expert pathologist. Patients undergoing ileocolic resection for other conditions (e.g. right colon cancer) and with healthy terminal ileum were used as controls. For each region (normal CD, inflamed CD, fibrotic CD and control), immunohistochemistry (IHC), RNA and protein evaluations for PAD2 and PAD4 were performed. Multiplex immunofluorescence (IF) for PAD2, PAD4, myeloperoxidase, neutrophil elastase, CD68, vimentin and α-smooth muscle actin were carried out to investigate the enzymes-expressing cells. Additional IF was performed to study citrullinated histone 3 (H3cit) expression, the product of PAD4 activity in neutrophils and component of neutrophil extracellular traps (NETs). Statistical analysis was carried out with Kruskal–Wallis test and post hoc Mann–Whitney test.

Results

Resection specimens from 13 CD and 11 controls were included. IHC and IF showed an increased expression of both PAD2 and PAD4 in the neutrophils of inflamed areas, in cytoplasm and nucleus, respectively (Figure 1). Activated fibroblasts (vimentin+ and α-smooth muscle actin+) were negative for both enzymes. PAD4 mRNA expression was increased in inflamed tissue (p = 0.001, p = 0.008 and p = 0.028 vs. normal CD, fibrotic CD and controls, respectively), and confirmed using Western Blot (Figure 2). H3cit was increased in the ileal inflammatory infiltrates too (Figure 3), confirming high PAD4 expression. For PAD2, no significant changes were observed at RNA and protein level, mainly due to its reduced expression in epithelial cells from normal to diseased tissue (Figure 4).

Conclusion

Both PAD2 and PAD4 are strongly expressed in neutrophils of CD ileal resection specimens, but only PAD4 shows a significantly higher expression in the inflammatory context which translates in the formation of NETs. No direct correlation was observed between PAD enzymes and intestinal fibroblasts.
P068 Erythrocyte and reticulocyte parameters in children with inflammatory bowel diseases
Authors: Semikina E , Potapov A , Tsvetkova V , Anoushenko A , Akulova S
Published: January 2020

Background

Anaemia is a frequent extraintestinal manifestation of inflammatory bowel diseases (IBD) and its severity leads to a deterioration in patient’s condition. Timely diagnosis and pathogenetic therapy are important for the effective treatment. The aim of this study was to evaluate the current cell indexes obtained from automatic blood analyser to identify the pathogenetic features of anaemia in children with IBD.

Methods

A total of 1400 blood samples were analysed in 420 children with IBD (177—Crohn’s disease (CD) and 243—ulcerative colitis (UC)) followed up in our centre in 2018–2019. A blood test was performed by flow hemocytometry on an automatic blood analyser Sysmex XN 1000. The evaluated parameters were: MCV, MCH, MCHC, RET, IFR and RET-He. Anaemia was established with haemoglobin level <110 g/l. Clinical activity was evaluated according PUCAI (UC) and PCDAI (CD) scores.

Results

Anaemia was detected in 31.7% of patients with UC and in 26.8% of patients with CD. Decreasing of Hb level was observed in both diseases with an increasing in the clinical activity. The average Hb levels in CD in remission, moderate and severe activity were 132.15 ± 17.07, 120.51 ± 18.28 and 105.06 ± 12.46 g/l, respectively; in children with UC with remission, minimal, moderate and severe activity 124.86 ± 16.01; 114.53 ± 15.56; 109.71 ± 20.34 and 96.09 ± 16.52 g/l, respectively. In patients with CD, anaemia was predominantly hypochromic: the average values of MCV, MCH and RET-He indicates a leading role the bioavailable iron deficiency and corresponded to those in CD remission: 82.8 ± 7.0 fl, 27.35 ± 3.02 pg and 31.28 ± 3.82 pg; in low/moderate CD activity: 78.93 ± 7.49 fl, 25.29 ± 3.51 pg, 28.62 ± 5.1 pg; in high CD activity: 74.7 ± 6.74 fl, 22.99 ± 2.69 pg, 27.14 ± 4.99 pg, respectively. RET count in CD was moderate and equal 58–68 cell/ml. With an increase in the UC process activity, hypochromia was not enhanced: MCV in the range 79–82 fl, MCH—25–26 pg. RET-He in remission of UC were 30.22 ± 3.95 pg; in minimal activity—28.93 ± 4.96 pg; in moderate activity—27.99 ± 4.62 pg and only with high activity of UC there was a decreasement to 25.21 ± 4.99 pg. In UC the level of RET and IFR increases with increasing severity of the disease: 57.75 ± 17.48‰, 63.31 ± 31.12‰, 77.29 ± 41.81‰, 111.29 ± 78,9‰ and IFR 11.62 ± 5.235%, 15.69 ± 8.065%, 18.97 ± 8.385%, 26.97 ± 9.785%, respectively. This suggests that anaemia in UC is mainly associated with blood loss.

Conclusion

Anaemia in different forms of IBD in children has different pathogenetic mechanisms. Evaluation of current parameters of red blood cells by modern automatic blood count analyser is cheap and informative for the differential diagnosis of anaemic syndrome in children with IBD.
P070 Machine learning approaches to identify IBD biomarkers from longitudinal microbiome data
Authors: Madgwick M , Sudhakar P , Tabib N S , Norvaisas P , Creed P , Verstockt B , Vermeire S , Korcsmáros T
Published: January 2020

Background

Inflammatory bowel disease (IBD) has been shown to be associated with alterations in the intestinal microbiome. However, the precise nature of these microbial changes remains unclear. With billions of microbes present within the gut, novel and powerful computational techniques are required to identify the relevant shifts in microbiota contributing to the disease. Machine learning (ML) allows a data-driven approach to identify these discrete dynamic changes, while the findings of the ML algorithms can be interpreted using systems biology (SB) techniques. By combining ML and SB approaches, we aim to characterise key microbial factors in IBD pathogenesis, distinct patterns of variability in a diverse patient cohort and provide a method for patient stratification.

Methods

The causal relationship between the changes in the gut microbiome and IBD is difficult to establish. Data from cross-sectional studies are plagued by confounding factors and inconsistencies between cohorts. To overcome this, the authors used rich longitudinal datasets and integrated metagenomic, multi-omic and clinical patient data. This workflow has been validated using large longitudinal IBD databases, including data from IBDMDB. We assessed the performance of the ML models using well-documented performance metrics to ensure the outcomes were robust.

Results

As a baseline, we used multiple ML models to predict disease type (UC, CD and non-IBD) from integrated multi-omics profiles. We analysed multiple ML techniques, including linear (e.g. linear mixed model), non-linear (e.g. Random Forest), time-series models (e.g. Rotation Forest) and deep learning models (e.g. long short-term memory network model). The authors identified the models which would allow flexibility to analyse the dynamic nature of the microbiome and allow integration of the microbiome data with clinical patient data. The payoff of greater flexibility was a reduction in the model performance in terms of identifying specific features from the metagenomics that could be used as biomarkers. However, we were able to identify connections between microbial and host proteins relevant to IBD and were able to stratify these by the patient’s metagenomic data.

Conclusion

We have developed an integrated ml-based microbiome analysis pipeline to identify biomarkers for IBD from longitudinal metagenomic data. Furthermore, using a variety of SB approaches, we were able to interpret the predicted key microbial features and communities by inferring connections between microbial and host proteins. This pipeline will enable us to analyse vast amounts of patient microbiome data in the context of clinical and metagenomic data, to allow identification of biomarkers for disease subtypes.
P075 Muc5ac expression protects the colonic barrier in experimental colitis
Authors: O’Connell L , Olli K , Rapp C , Collins C , McNamee E , Jensen O , Jedlicka P , Allison K , Goldberg M , Gerich M , Evans C , AHERNE C
Published: January 2020

Background

The mucus gel layer (MGL) lining the colon is integral to exclusion of bacteria and maintaining intestinal homeostasis in health and disease. MGL defects allowing bacteria to directly contact the colonic surface are commonly observed in ulcerative colitis (UC). The major macromolecular component of the colonic MGL is the secreted gel-forming mucin, MUC2, whose expression is essential for homeostasis in health. In UC, another gel-forming mucin, MUC5AC is induced. In mice, Muc5ac is protective during intestinal helminth infection. Here, we tested the expression and functional role of MUC5AC/Muc5ac in colitis patient biopsies and murine colitis.

Methods

We measured MUC5AC/Muc5ac expression in UC patient biopsies and during acute dextran sulphate sodium (DSS) colitis. We performed DSS-colitis in mice deficient in Muc5ac (Muc5ac−/−) to model the potential functional role of Muc5ac in colitis. To assess MGL integrity, we quantified bacterial–epithelial interaction and translocation to mesenteric lymph nodes (MLNs). Antibiotic treatment was performed to directly investigate the role of colonic bacteria in our murine colitis studies.

Results

Colonic MUC5AC/Muc5ac mRNA expression increased significantly in active UC and murine colitis. Muc5ac−/− mice experienced worsened injury and inflammation in DSS-colitis compared with controls. This was associated with increased bacterial–epithelial contact and translocation to the MLN. Antibiotic treatment normalised colitis severity in Muc5ac−/− mice to that of antibiotic treated controls.

Conclusion

We demonstrate for the first time that MUC5AC/Muc5ac induction in acute colitis controls injury by reducing bacterial breach of the MGL. Therefore, developing strategies to induce MUC5AC expression may protect the intestinal barrier in UC.
P080 Therapeutical effect of urine-derived stem cells on DSS-induced chronic model of mice
Authors: Wu X R , Zhou C , Liu H S , Xuan-hui L , Hu T , Zheng X B , Liang Z X , Wu X J , Gao X , He X S , Ke J , Chen Y F , He X W , Hu J C , Zou Y , Lan P
Published: January 2020

Background

The application of stem cell therapy in the treatment of inflammatory bowel diseases (IBD) is limited because of the invasive approaches of stem cells. Urine-derived stem cells (USCs) were recently shown to have regenerative properties, which can be harvested in a safe, low-cost and non-invasive way.

Methods

Human USC were isolated and expanded from the urine of healthy male adult volunteers (n = 3, age arrange 24–30 years old). USC were characterised by cell surface marker expression profile and multipotent differentiation. In vivo therapeutic value of USC was assessed using murine colitis chronic model induced by dextran sulphate sodium (DSS).

Results

USC were positive for mesenchymal stem cell markers but were negative for hematopoietic stem cell markers. These cells differentiated into osteo-, adipo- and chondro-genic cell lineages. Systemic administration of USC significantly ameliorated the clinical and histopathological severity of colitis and increased the survival rate in chronic murine colitis model.

Conclusion

This study demonstrated that implantation of USC reduces inflammation in IBD rodent model, indicating that USC therapy serves as a potential cell-based therapeutic candidate for IBD.
P087 Biomarkers of elastin degradation differentiate between clinically inactive and active disease in ulcerative colitis patients
Authors: Pehrsson M , Domislović V , Karsdal M A , Brinar M , Barisic A , Krznaric Z , Manon-Jensen T , Mortensen J H
Published: January 2020

Background

In ulcerative colitis (UC), the state of chronic inflammation results in increased matrix metalloprotease (MMP) and serine protease activity, which effectively leads to a higher degree of intestinal tissue remodelling, including components of the extracellular matrix (ECM). One of these components is elastin a matrix protein of the interstitial matrix in the lamina propria and submucosa, providing tissue resilience and elasticity. As such, we investigated whether elastin degradation in UC patients was associated with disease activity and severity, potentially enabling patient differentiation based on elastin degradation.

Methods

Twenty-nine UC patients and 29 healthy donors were included in the study. Disease activity was determined according to the partial Mayo score (pMayo >1) and the Mayo Endoscopic Score (MES). Disease severity and extension was assessed using the Montreal classification. Disease severity was additionally assessed using the Trulove and Witt’s (TW) clinical score. The biomarkers of elastin degradation included: MMP-7 (ELM-7) cathepsin-G (EL-CG) and proteinase-3 (ELP-3), measured in serum by ELISA. One-way ANOVA (Kruskal–Wallis) correcting for the false discovery rate were applied for the statistical analysis.

Results

TW: ELP-3 levels in moderate-to-severe UC patients were significantly elevated in comparison with HD (p < 0.001). Partial Mayo: EL-CG levels in patients with active UC were significantly elevated in comparison with HD (p < 0.01), and UC patients in remission (p < 0.01). ELP-3 levels were likewise significantly elevated in active UC patients compared with HD (p < 0.001), and UC patients in remission (p < 0.01). Montreal classification: ELM-7 was significantly elevated in active UC compared with HD (p < 0.05), and UC patients in remission (p < 0.05). EL-CG were also significantly elevated in active UC compared with HD (p < 0.05), and UC patients in remission (p < 0.05). ELP-3 was significantly elevated in active UC compared with HD (p < 0.01). According to the MES score, ELP-3 levels in moderate-to-severe UC patients were significantly elevated in comparison to HD (p < 0.01).

Conclusion

The data presented in this study demonstrate an association between biomarkers of proteolytic elastin degradation and disease activity in UC patients especially the protease-3-derived biomarker, ELP-3, showed significant association with active UC in all the clinical scoring systems as well as the MES score. Utilising these minimally invasive elastin degradation biomarkers could serve as surrogate markers for monitoring of disease activity and potentially aid the differentiation of patients with an active disease from patients in remission or with a lower disease activity for UC.
P088 TLR3 ameliorates colitis-associated colon tumourigenesis in mice
Authors: Jun Y K , Koh S J , Gang H W , Chung K Y , Han J M , Kim J W , Kim B G , Lee K L , Im J P , Kim J S
Published: January 2020

Background

Toll-like receptor 3 (TLR3) signalling regulates innate and adaptive immune systems by the recognition of dsRNA. Activation of TLR3 signalling by poly(I:C) attenuates dextran sodium sulphate (DSS)-induced murine colitis. However, little information is available on the role of TLR3 signalling in the development of colitis-associated colon tumourigenesis.

Methods

Wild-type (WT) and TLR3-deficient (TLR3−/−) mice were intraperitoneally injected azoxymethane (AOM) 12.5 mg/kg on day 0 followed by three cycles of 2% DSS for 5 days and 2 weeks of free water consumption. Clinical indices such as weight change, colon length, the number of tumours, and the histologic severity of colitis were evaluated in each experiment. Immunohistochemical or immunofluorescence analyses for phospho-IκB kinase (IKK) and β-catenin were performed in colon tissues. To elucidate the antitumourigenic mechanism by colon inflammation, poly(I:C) or PBS was intraperitoneally injected in the AOM/DSS-induced tumourigenesis model in WT mice. To evaluate direct antitumor effect on tumourigenesis, as first experimental model, both WT and TLR3−/− mice were intraperitoneally injected AOM weekly for 12 weeks without DSS treatment. As the second experimental model, WT and TLR3−/− mice were received 2% DSS mixed with drinking water three times for 5 days every 2 weeks after one intraperitoneal AOM injection.

Results

TLR3−/− mice exhibited a higher tumour burden compared with wild-type mice. Body weight loss was greater in TLR3−/− mice than in WT mice. However, here was no significant difference in colon length and the severity of colitis between the two groups. Immunoreactivity for β-catenin was markedly increased in TLR3−/− mice. However, there was no difference in IKK expression. Activation of TLR3 by poly(I:C) was not associated with the reduced tumour development in WT mice. However, repeated AOM injections without DSS resulted in greater body weight loss in TLR3−/− mice than in WT mice, which was associated with the increased tumour development in TLR3−/−mice.

Conclusion

TLR3 signalling attenuated colitis-associated colon cancer development. Based on our experiments, TLR3 signalling inhibits colon tumourigenesis by direct antitumor activity rather than anti-inflammatory effect of colitis.
P094 Differences in expression of PPARγ in small intestine vs. colon impact the effect of 5-aminosalicylates in inflammatory bowel disease
Authors: Bakke I , Bruland T , Eriksen K S , Skovdahl H K , Gopalakrishnan S , Thorsvik S , Granlund A V B , Sandvik A K , Østvik A E
Published: January 2020

Background

The nuclear receptor peroxisome proliferation-activated receptor γ (PPARγ) harbours anti-inflammatory effects. There is evidence that PPARγ mediates the effect of 5-aminosalicylic acid (5-ASA). 5-ASA is the first-line drug in ulcerative colitis (UC, while its use in Crohn`s disease (CD) is debated and not recommended according to guidelines. We hypothesise that the inconsistent therapeutic effect of 5-ASA in Crohn’s disease is caused by different expression of PPARγ between the large and small intestine.

Methods

Levels of PPARγ mRNA were measured by RNASeq in mucosal biopsies from (1) active/inactive ileal CD (n = 14/17) and healthy controls (n = 9) and (2) colonic biopsies from patients with active/inactive UC (n = 24/24) and active/inactive CD (n = 24/21) and healthy controls (n = 20). Subsets of ileal and colonic biopsies were examined by western blot, immunohistochemistry (IHC) and in situ hybridisation (ISH). The effects of 5-ASA on PPARγ expression and TNF/IL17/polyI:C induced cytokine release were examined in the colonic cell-line HT29 and primary human IECs (colonoids) using RNASeq and ELISA in.

Results

PPARγ mRNA was strongly downregulated in colonic biopsies from inflamed mucosa of UC (log2 = −1.65, fold change 0.319 (p < 0.001)) and CD (log2 = −1.3, fold change 0.406 (p < 0.001)) compared with healthy controls. In ileal biopsies from CD and controls, PPARγ mRNA was not differentially expressed between inflamed and non-inflamed or healthy mucosa. These findings were confirmed by ISH in a subset of biopsies. Western blot analysis and IHC revealed almost undetectable levels of PPARγ protein in ileum, in the colon however, PPARγ was strongly expressed in healthy controls and inactive IBD; with significantly lower levels in active IBD. PPARγ was downregulated by TNF, TNF+IL17 and TNF + poly(I:C) in colonoids derived from IBD-patients (n = 3) and non-IBD controls (n = 3). 5-ASA attenuated the release of TNF-induced proinflammatory cytokines such as CXCL1, CXCL8 and CXCL10 from both HT29 cells and colonoids. This effect was reversed by the PPARγ antagonist GW9662.

Conclusion

5-ASA harbours anti-inflammatory effects on intestinal epithelial cells mediated by epithelial PPARγ. We suggest that the previously observed lack of effect of 5-ASA in CD is related to differences in PPARγ expression in small intestine vs. colon. These results suggest that patients with Crohn’s colitis may benefit from 5-ASA similarly to UC patients and challenge the current view on use of 5-ASA in CD.
P096 Specific plasma micro-RNA profiles for prediction of recurrence in Crohn’s disease in pre-surgery and established recurrence scenarios
Authors: Moret I , Cerrillo E , Saez-Gonzalez E , Hervas D , Iborra M , Tortosa L , Nos P , Gadea J , Beltran B
Published: January 2020

Background

Crohn’s disease (CD) patients who undergo ileocolonic resection are at high risk of postoperative recurrence (POR). Both environmental and genetic factors are known to influence CD pathogenesis. MicroRNAs (miRs) are important epigenetic regulators in CD, but their role in the progression to POR is still unknown. Thus, we aimed to identify differential miRs profiles and their potential use to predict POR in CD patients.

Methods

A cohort of 33 CD patients who underwent a resection was consecutively and prospectively followed for at least 1 year. Patients were classified according to the presence or absence of POR during the follow-up. POR was assessed by ileocolonoscopy (Rutgeerts’ index ≥ i2b) or MRI enterography within 6–12 months after surgery. Patients received postoperative therapy to prevent POR (thiopurines 47%; anti-TNFα 25%; mesalamine 9%; combination therapy 3%; none 16%). Peripheral blood samples were taken at fixed times. Total RNA was obtained from miRNeasy Serum/Plasma Advanced Kit (Qiagen) and run on the miRCURY LNAtm Universal RT microRNA PCR with a miRs panel selection (32) based on previous experiments. The predictive accuracy of the possible predictors was assessed by using receiver operating characteristic curves with the R software.

Results

MicroRNAs profiles were studied at two different times: presurgery (PS) and at the time of the morphological POR or, in those who remained in remission, 1 year after surgery (1 year). The miRs whose fluorescent signal did not differ from background noise were excluded from the analysis [8 in PS (25%); 5 in 1 year (16%)]. Using elastic net regression, five miRs (hsa.miR.191.5p, hsa.miR.15b.5p, hsa.miR.106b.5p, hsa.miR.451a and hsa.miR.93.5p) were selected for discriminating the two patient groups at the time pre-surgery. Seven miRs (hsa.miR.15b.5p, hsa.miR.126.3p, hsa.miR.451a, hsa.let.7b.5p, hsa.miR.93.5p, hsa.miR.423.5p and hsa.miR.125b.5p) were selected for discriminating the two patient groups at the time of 1 year. Using a predictive model with these miRs, an overall AUC of 0.86 (95% confidence interval, 0.75–0.97) was achieved for PS, and 0.96 (0.92–1.0) for 1 year.

Conclusion

This study shows that plasma microRNAs are useful biomarkers in the POR scenario. When recurrence is already established, a group of plasma miRs is also specifically expressed, thus assessing them could prevent patients from receiving and endoscopic procedure for POR evaluation. Furthermore, microRNA assessment PS could predict which patients will develop recurrence within 1 year of surgery. This could also have an impact on clinical management, selecting patients who can avoid immunosuppressive treatment and the ones who may need more intensive therapeutic strategies.
P103 The treatment effect of tonsil-derived mesenchymal stem cells (T-MSCs) via regulating programmed death-1/programmed death ligand-1 (PD-1/PD-L1) in ulcerative colitis
Authors: Song E M , Joo Y H , Lim J Y , Choe A R , Tae C H , Moon C M , Kim S E , Jung H K , Shim K N , Jung S A
Published: January 2020

Background

The programmed death 1 (PD-1)/programmed death-ligand 1 (PD-L1) pathway has not been fully evaluated in inflammatory bowel disease. We evaluated the level of PD-1/PD-L1 in the mucosa of patients with ulcerative colitis and revealed the role of PD-1/PD-L1 in the immunomodulatory mechanism of tonsil-derived mesenchymal stem cells (T-MSCs).

Methods

We used chronic murine colitis model induced by dextran sodium sulphate (DSS). We measured the PD-1 and PD-L1 levels in inflamed colonic tissues before and after the treatment with T-MSC. We also measured the levels of PD-1 and PD-L1 in the colonic tissues from patients with ulcerative colitis and compared with those from normal controls.

Results

In chronic colitis model, the level of PD-L1 was decreased than normal controls (fold change) (1.0 vs. 0.46 ± 0.08, p = 0.05). However, the level of PD-1 was increased than normal controls (1.0 vs. 6.84 ± 8.16, p = 0.142). After treatment with T-MSC which showed significant improvement in body weight, disease activity index and colon length, the levels of PD-1 and PD-L1 were recovered; PD-L1 was significantly increased (1.04 ± 0.77 vs. 0.46 ± 0.08, p = 0.031) and the level of PD-1 was decreased (3.63 ± 1.94 vs. 6.84 ± 8.16, p = 0.537). When measuring the level of PD-1 and PD-L1 in both soluble medium and cell lysate of T-MSC, we found that the PD-1 and PD-L1 were expressed in both forms (182.63 pg/ml in soluble medium and 11.85 pg/protein (μg) in cell lysate after 7 days of differentiation period). In the analysis using human colonic tissues, a significantly increase in the levels of PD-1 and PD-L1 was observed in the colonic mucosa of patients with UC compared with normal controls (PD-1: 2.09 ± 3.95 vs. 7.93 ± 14.21, p = 0.034 and PD-L1: 2.24 ± 2.91 vs. 10.08 ± 18.13, p = 0.044).

Conclusion

The altered expression of PD-1 and PD-L1 in colonic mucosa may a possible mechanism of ulcerative colitis and T-MSC-derived PD-L1 could repress the colitis.