bluuebunny/crossref_metadata_2025_split huggingface dataset:
P066 Azathioprine linked with impaired intestinal epithelial regeneration in Crohn’s disease
Authors: Borycka-Kiciak K , Pietrzak A , Ferenc K , Pietrzak P , Janaszek L , Tarnowski W , Zabielski R 
Published: January 2020

Abstract

Background
In patients operated due to active Crohn’s disease (CD), the negative effect of steroids and biologics on septic complications is known. Until now, azathioprine (AZA) is considered to be safe. The aim of our study was to assess the impact of AZA on intestinal epithelial cells damage in patients with active CD as a surrogate marker of healing.

Methods
Intestinal specimens taken from macroscopically healthy surgical margins of all consecutive CD patients operated due to active isolated ileocecal disease during the study period (2014–2016) were evaluated. Expression of caspase-3, p-53 and Ki-67 as markers of cell apoptosis, DNA damage and proliferation were immunohistochemically tested and assessed using a confocal microscope and microimage for in-tissue-cytometry analysis. Western-blot analysis was performed for the evaluation of cellular integrity using ZO-1 and E-cadherin as a markers. 30-day clinical outcomes were assessed. The study was approved by the institutional Ethics Committee.

Results
From 61 operated due to active CD patients, 35 met the inclusion criteria. Patients were divided accordingly to preoperative treatment: treated with no immunomodulators (N-9 patients), on steroids (S-14 patients), on AZA (A-6 patients), and on combination therapy, AZA + steroids (AS-6 patients). There were no substantial differences between groups. We found statistically significant increase of apoptosis in A group compared with N (5.33 ± 1.05 vs. 1.29 ± 0.51, p = 0.011), but also S group (1.58 ± 0.68, p = 0.014) and increase of DNA damage in A, AS and S groups compared with N group (p =0.01; p = 0.032; p = 0.035, respectively) (Figure 1a–c). P53-mediated cell cycle arrest and apoptosis through a caspase-3-dependent pathway in response to DNA damage was the most intensive in A group (Figure 2a). Reduction of cell proliferative activity in group A did not reach statistical significance (p = 0,057). A reduction of ZO-1 in A group and increased level of E-cadherin in S group were found. The effect of the decreased number of tight junctions and disintegration of the mucosa layer was observed in A group (Figure 2b). Clinically, in 30-day postoperative follow-up, six wound healing complications and one anastomotic leak were found, all in patients treated with immunomodulators.

Conclusion
We found that in epithelial cells of the small and large intestine of patients treated with AZA, apoptotic activity and DNA damage processes are increased when regeneration processes and mucosal integrity are significantly disturbed. These abnormalities of intestinal epithelial regeneration may be a surrogate marker of impaired mucosal healing.
P084 Cannabis use promotes lymphocyte responsiveness and modifies T-cell signalling in adolescent IBD patients
Authors: Sanctuary M , Wilkinson C , Jones A , Murphy B , Hoffenberg E , Collins C 
Published: January 2020

Abstract

Background
The prevalence of inflammatory bowel disease (IBD) has increased dramatically in recent years, particularly in paediatric populations. The efficacy of available therapies is limited and often transient, leading patients to seek alternative therapies for symptom relief, including the use of medical marijuana (Cannabis sativa). There is some evidence to suggest that cannabinoids exert anti-inflammatory effects in preclinical colitis models. In addition, a number of small studies have reported improvements in non-empirical disease measures and reductions in visceral pain in response to cannabis use. However, associations between cannabis use in IBD and increased need for surgical interventions have also been reported.

Methods
Therefore, determining the direct impact of cannabis use on immune modulation in IBD patients is of critical importance. Blood samples collected from paediatric IBD patients who reported cannabis use for symptom control were analysed for cytokine expression and T-cell signalling pathway activation compared with non-users. Concentrations of serum phytocannabinoids were determined by HPLC and correlated with subsequent ex vivo assay parameters.

Results
Results demonstrated elevated levels of a myriad of pro-inflammatory cytokines in users vs. non-users upon ex vivo restimulation. This coincided with an expansion of pathogenic Th17 cells in the periphery. Differences in signalling cascades of activated T cells between users and non-users were also observed.

Conclusion
These results suggest that cannabis exposure, which can desensitise cannabinoid receptors, may prime pro-inflammatory pathways in paediatric IBD patients. Future studies should address the limitations of observational studies through the use of randomised controlled trials of cannabis use in paediatric IBD populations.
This research was funded by the Colorado Department of Public Health and Environment.
P090 Influence of high-fat diet on colitis and liver health in inflammatory bowel disease: a study using murine model of DSS-induced colitis
Authors: Choe A R , Kim S E , Lee H A , Yoo S G , Kim T O , Tae C H , Moon C M , Jung H K , Shim K N , Jung S A 
Published: January 2020

Abstract

Background
In recent, consumption of high-fat (HF) diet has been increasingly considered to be a significant aggravating factor of inflammatory bowel disease and non-alcoholic steatohepatitis. The aim of this study was to investigate whether exacerbation of chronic DSS-induced colitis caused by HF diet could promote liver inflammation, using measurement of biomarkers.

Methods
Male C57BL/6 mice were allocated into four experimental groups receiving either; (1) control with normal diet/water; control, (2) control with high-fat diet; HF, (3) DSS induced colitis group with normal diet/water; DSS, (4) DSS-induced colitis group with high-fat diet; DSS + HF for 4 weeks. On day 30 of study, colon and liver tissues samples were obtained for measuring mRNA expression of pro-inflammatory/pro-fibrogenic markers and blood for metabolic parameter analysis. Spearman rank correlations were analysed between colon and liver inflammatory biomarkers.

Results
The mean colon length was shorter, and more stenotic lesions were observed in DSS + HF group, compared with the other three groups. While HF diet alone did not cause significant colonic inflammation, HF diet in chronic DSS colitis aggravated colonic inflammation revealing the significant increase of IL1-β and IL17 mRNA expression as well as increasing the disease activity indices significantly at the late phase of the study. Moreover, DSS + HF group showed significantly higher serum levels of total cholesterol (p = 0.004) and glucose (p = 0.002), compared with HF group. Hepatic inflammation was more severe in DSS + HF group, showing significantly increased IL1-β and TGF-β expression compared with HF group. The mRNA expression of IL-1β in colitis group was statistically positive correlated with those in liver tissue (rs = 0.51, p = 0.010).

Conclusion
High-fat diet in chronic colitis may adversely affect the metabolic parameters and liver inflammation, as well as aggravation of colitis.
P091 Dietary triggers of colonic inflammation following treatment with exclusive enteral nutrition in children with Crohn’s disease
Authors: Gkikas K , Logan M , Nichols B , Clark C M , Ijaz U Z , Gervais L , Duncan H , Garrick V , Curtis L , Buchanan E , Cardigan T , Armstrong L , Delahunty C , Flynn D M , Barclay A R , Tayler R , Hansen R , Russell R K , Gerasimidis K 
Published: January 2020

Abstract

Background
Exclusive enteral nutrition (EEN) ameliorates gut inflammation in children with Crohn’s disease (CD). We have previously described the rapid rise in faecal calprotectin levels (FC) when children with CD return to their habitual diet after EEN treatment (1). We performed dietary assessment and measured biomarkers of food consumption in faeces of children with CD during early food reintroduction.

Methods
A composite outcome of clinical remission post EEN (weighted Paediatric Crohn’s Disease Activity Index &lt;12.5) and a significant drop in FC (FC decrease: &gt;500 mg/kg / &gt;35%) was used to define the patient group. All patients completed 3-day estimated weight food diaries and provided a faecal sample. Patients were divided equally for statistical analysis purposes in two groups; above (Group 1) and below (Group 2) the median FC concentration at food reintroduction. Nutrient analysis was performed with WinDiets. Short-chain fatty acids, the gluten immunogenic peptide (GIP) and starch were measured in stool, as proxy of fibre, gluten and malabsorbed starch, respectively. Food groups were assigned using the National Diet and Nutrition Survey approach and statistics employing taxonomy were utilised in the food group analysis. All data are displayed using medians (Q1, Q3).

Results
Fourteen children provided an FC sample within 21 (15, 51) days post EEN. Patients were of similar age, weight and height (p = 0.4, p = 0.37, p = 0.79). Total energy intake did not differ between the groups (p = 0.37). Patients in Group 1 consumed more fibre than Group 2 [12.1 g (11.3, 19.9) vs. 9.9 g (7.6, 12.1), p = 0.04]. Protein and phosphorus intakes, expressed as a percentage of reference nutrient intakes, were also higher in Group 1 than Group 2 [protein (%): 262 (195, 291) vs. 211 (88, 215), p = 0.02], [phosphorus (%): 241 (171, 290) vs. 177 (131, 1901), p = 0.04]. Butyrate and valerate levels were higher in Group 1 than Group 2 [butyrate: 62.5 (41.8, 114) vs. 34.1 (29.2, 49.6), p = 0.02; valerate: 13.7 (9.3, 17.9) vs. 4.5 (2.3, 8.1), p &lt; 0.01. There were no differences in GIP and faecal starch levels between the two groups. Red and processed meat intake was higher in Group 1 than Group 2 [151 g (66.7, 190) vs. 63.3 g (21.7, 67), p = 0.02)]. Cereals intake was non-significantly higher in Group 1 [389 (207, 405) vs. 231 (141, 279), p = 0.08]. Overall diet diversity did not differ between the two groups (p = 0.47).

Conclusion
The current analysis suggests that fibre, protein, phosphorus and red and processed meat may be associated with recurrence of colonic inflammation in children with CD during early food reintroduction. These findings should be confirmed in larger studies.

Reference
P093 Crosstalk between Toll-like receptor 4 and enteric serotonergic pathways in a mouse model of dinitrobenzene sulfonic acid-induced colitis
Authors: Cerantola S , Ridolfi M , Faggin S , Tegon M , Bussolaro G , Salviato E , Bistoletti M , Caputi V , Giaroni C , Giron M C 
Published: January 2020

Abstract

Background
Changes in serotonin (5-HT) levels, anomalies in serotonergic and cholinergic machinery and altered Toll-like receptor 4 (TLR4) expression have been shown in IBD in patients and related animal models. Thus, we aimed to assess the crosstalk of enteric serotonergic system and TLR4 signalling in a mouse model of dinitrobenzene sulfonic acid (DNBS)-induced colitis.

Methods
Male C57/Bl6 (WT) and TLR4−/− mice (9 ± 2 weeks old; N = 10 mice) were presensitised with 1% dinitrobenzene sulfonic acid (DNBS), and after 1 week was intrarectally instilled with 2.5% DNBS. Small intestine inflammation was measured by disease activity index and histological analysis. Changes in ileal muscle tension were isometrically recorded following: (1) cumulative addition of carbachol (CCh; 0.1–100 µM); (2) electric field stimulation (EFS, 0–40 Hz); (3) 60 mM KCl; (4) 30 μM 5-HT addition with or without 0.1 μM ondansetron (5-HT3R antagonist). Immunofluorescence distribution of the neuronal HuC/D and nNOS and glial GFAP markers were determined in longitudinal-muscle-myenteric plexus whole mounts (LMMPs) by confocal microscopy.

Results
In WT mice, DNBS treatment altered receptor and not-receptor mediated responses (+120% of Emax to CCh and +103% of contraction to KCl, respectively; p &lt; 0.001, N = 5 mice/group) together with an altered cholinergic neurotransmission (−50% at 10 Hz; p &lt; 0.01, N = 5 mice/group) and 2-fold increase to 30 μM 5-HT-mediated response (p &lt; 0.001, N = 5 mice/group). After DNBS treatment TLR4−/− mice showed a significant increase in excitatory-mediated response (+98% of Emax to CCh; +80% of contraction to KCl; +120% at 10 Hz; p &lt; 0.001, N = 5 mice/group) together with a significant reduction of 30 μM 5-HT-mediated response (−50%, p &lt; 0.001, N = 5 mice/group). These changes were associated to a significant decrease of the total number of HuC/D+ neurons (−44% and −19% for WT DNBS and TLR4 DNBS mice, respectively) together with a 1.3-fold increase in S100b immunofluorescence in WT mice after DNBS treatment.

Conclusion
These findings not only suggest an important role of TLR4 in small intestine neuromuscular dysfunction during colitis but also provide novel information on the potential benefits of targeting TLR4 in various gut disorders that exhibit aberrant cholinergic and 5-HT signalling.
P098 Receptor-interacting protein 1 kinase inhibition therapeutically ameliorates experimental T-cell-dependent colitis in mice
Authors: Gobbetti T , Kathryn F , Beal A , Rowles A , Pearse G , Harada I , Bertin J , Haynes A , Berger S 
Published: January 2020

Abstract

Background
Inflammatory bowel disease (IBD) is a multifactorial disorder characterised by chronic and relapsing intestinal inflammation. Receptor-interacting protein 1 (RIP1) kinase activity is emerging as a driver of pro-inflammatory cytokine production and cell death and has been implicated in multiple inflammatory diseases including IBD. To this end, recent work has shown that RIP1 kinase inhibition reduces the spontaneous production of cytokines from human ulcerative colitis (UC) and Crohn’s disease explants.

Methods
The aim of this study was to investigate the anti-inflammatory effect of a highly selective inhibitor of RIP1 kinase activity (GSK547A) in the mouse T-cell transfer model of colitis; a model that shares many features with human IBD. All animal studies were ethically reviewed and carried out in accordance with Animals (Scientific Procedures) Act 1986 and the GSK Policy on the Care, Welfare and Treatment of Animals. Chronic colitis was achieved by transferring CD4+CD45RBhigh T cells into immunodeficient female SCID mice (6/8 weeks old). After confirming the development of pathology using endoscopy and MRI, animals were treated therapeutically with the RIP1 inhibitor GSK547A (50 mg/kg twice a day PO) or vehicle (0.5% hydroxypropyl methylcellulose in water). The severity of colitis was measured 5 weeks after cell transfer both macro- and microscopically. Mucosal damage by endoscopy was also assessed. RT-PCR and MSD analysis were performed to detect colon cytokine/chemokine and calprotectin levels. SAA in the plasma was measured using ELISA.

Results
We found that treatment with GSK547 significantly ameliorated experimental T-cell-dependent colitis in mice. GSK547A-treated mice displayed decreased weight loss, colon density (ratio weight/length), macroscopic disease activity index, colon thickness compared with vehicle-treated animals. Mucosal damage, assessed by endoscopy and histopathology, was also reduced following treatment with RIP1 kinase inhibitor. In addition, GSK547A reduced the expression of cytokines in the colon (TNF-α, IL-17A, INF-γ, IL-6 and MCP-1), both at a protein and RNA level. Relevant translational biomarkers such as SAA in plasma and RNA calprotectin in the colon were also decreased in the GSK547A treated group when compared with vehicle.

Conclusion
Our results suggest that RIP1K inhibition is a strong protective factor with anti-inflammatory potential in the progression of chronic colitis, when applied to the translationally relevant T-cell transfer model of colitis. These findings suggest the potential application in the management of inflammatory bowel disease and support the ongoing clinical program evaluating the effect of RIP1 kinase inhibition in UC patients.
P104 Defective metabolism mediates NK cell dysfunction in inflammatory bowel disease (IBD)
Authors: Zaiatz Bittencourt V , Ryan E , Jones F , Doherty G 
Published: January 2020

Abstract

Background
Cell metabolism, in addition to providing energy and biochemical building blocks, also regulates immune cell function. We have increased our understanding of how natural killer cells (NK cell) metabolism impacts human NK cell function; however, little is known about NK cell role and metabolic changes in autoimmune diseases. The dysfunctional NK cell phenotype observed in pathological settings linked to defective cell metabolism will give insights into the development of new therapies for inflammatory diseases. IBD is an autoimmune disease of unknown origin characterised by severe gut inflammation and increased risk of colorectal cancer. NK cells can augment autoimmune responses through their activation of adaptive immune response by interaction with macrophages, dendritic cells and T cells; therefore, it is essential to decipher NK cells function in IBD to propose novel therapeutic targets. In this study, we defined the phenotype and metabolic profile of NK cells isolated from peripheral blood of IBD patients.

Methods
Blood samples were obtained from healthy donors and IBD patients from whom written consent had been obtained. PBMC were isolated by Lymphoprep (Axis-Shield) gradient. PBMC were stained with a combination of cell surface receptors, intracellular cytokines, proteins and analysed by flow cytometry. Cells were either stained ex vivo or after overnight stimulation with cytokine combination IL-12 + IL-15 and in the presence or absence of mTORC1 inhibitor rapamycin. Changes in OxPhos or glycolysis were detected by XFp extracellular ﬂux analyser (Agilent Technologies). Mitochondria mass and polarisation were measured by flow cytometry utilising MitoTracker Green and MitoTracker Red, respectively (ThermoFisher).

Results
NK cells from IBD patients have impaired mTORC1 activity (a key signalling pathway regulating cell metabolism), which is associated with increased expression of pro-inflammatory cytokines IL-17 and TNF-α. Patients NK cells have dysfunctional mitochondria characterised by reduced mitochondria mass, low membrane potential and limited OXPHOS and glycolysis. Furthermore, patients NK cells have limited IFNγ production, reduced TRAIL expression and defective capability of killing cancer cells.

Conclusion
Taken together, our results strongly imply that NK cells from IBD patients have a distinct bioenergetic programme that favours the increased production of pro-inflammatory cytokines. This might be an important unknown defect of NK cells in the regulation of inflammation in IBD that can be used for the development of new treatments. Further studies need to be conducted to identify the reason for the observed defects and determination of other key metabolic pathways involved in NK-cell function during IBD.
P107 The expression and regulation of Oncostatin M and its receptor in intestinal inflammation
Authors: Cineus R , Boesel D , Hainbuch S , Jukes C , Hsieh Y H , Reich J , Borek Z , Kuehl A , Hegazy A N 
Published: January 2020

Abstract

Background
Intestinal homeostasis depends on the interplay between the gut microbiota, epithelium and immune cells. A novel role of Oncostatin M (OSM), a pro-inflammatory cytokine has recently been identified in mouse and human intestinal inflammation. Previous studies have shown OSM as a key driver of chronic inflammation in anti-TNF-α-refractory colitis. A single-nucleotide polymorphism in the human OSM genetic locus is strongly associated with risk of developing inflammatory bowel disease (IBD), thus, biological therapies targeting OSM could have therapeutic potential. Our project aims to explore the impact of OSM on intestinal barrier function in health and disease.

Methods
To evaluate the role of OSM in intestinal inflammation, we utilized a combination of in vitro and in vivo techniques. This included the generation of 3D intestinal organoids from mice and patients. Organoids were stimulated with a repertoire of different cytokines to determine the responsiveness of OSM receptor (OSMR) to different cytokine signals using a quantitative-PCR-based approach. For in vivo modelling of disease, the Helicobacter hepaticus colitis model was used, as it combines both immune and dysbiosis-driven aspects of disease. This allowed us to measure OSM and OSMR expression in response to inflammation and within specific organs and cell subsets. Furthermore, RNAscope in situ hybridisation was used to determine the localisation of OSM- and OSMR-expressing cells in inflamed mucosal tissue from colitic mice and IBD patients.

Results
RNAscope in situ hybridisation as well as gene expression analysis have shown that the OSM and OSMR were highly expressed in C57BL/6 mice upon induction of colitis in the H. hepaticus model of disease and in mucosal tissues of IBD patients. In addition, a plethora of pro-inflammatory cytokines were upregulated during colitis, with colitic mice showing increased tissue pathology. Furthermore, FACS analysis shows excessive immune cell infiltration in the spleen, colon and mesenteric lymph nodes of colitic mice.

Conclusion
Our preliminary results have shown that different gut-resident hematopoietic and non-hematopoietic cell types express OSM and OSMR and this expression was modulated by pro-inflammatory cytokines. We therefore hypothesis that OSM might drive distinct transcriptional responses in various gut-resident cell populations. Thus, differential targeting of the OSM receptor might be a potential therapeutic approach in IBD.
P108 The importance of developing consensus-driven clinical data standards for Crohn’s disease to amplify data
Authors: Owen J , Baker R 
Published: January 2020

Abstract

Background
When CDISC standards are applied, data is collected, organised and analysed in a clear and consistent manner so that all researchers can leverage and share information from individuals and studies around the world. Required by the USA Food and Drug Administration (FDA) and Japan’s Pharmaceuticals and Medical Devices Agency (PMDA) and adopted by the world’s leading research organisations, CDISC standards enable the accessibility, interoperability, and reusability of data. Adopting CDISC standards is an invaluable investment that leads to more meaningful, more efficient research for your organisation and the entire global research community.

Methods
With support from The Leona M. and Harry B. Helmsley Charitable Trust and following the CDISC consensus-based standards development process, a team of Crohn’s disease and standards development experts was created to develop clinical data standards for Crohn’s disease. The CDISC standards development process consists of five stages:

Results
The project scoping stage completed in July 2019. The scoping team, consisting of Crohn’s disease clinical SMEs, identified the following topics that would most benefit from development of clinical data standards and to move into the concept modelling and standards development phases of the project:
Package 1
Package 2 Endoscopy assessments
The first package of data standards for Crohn’s disease will be available for Public Review in July 2020 and is anticipated to be published in Q1 2021 for use by researchers and industry. The second package of data standards will be available for Public Review in Q3 2020 with publication in Q2 2021.

Conclusion
CDISC’s vision is to encourage the global adoption of standards to leverage and realise the full potential of data and produce faster, more efficient research, paving the way for more breakthroughs in Crohn’s disease treatments, therapies and cures. ECCO attendees and the Crohn’s disease community are encouraged to participate in CDISC’s informational webinars and Public Review stages to foster all-inclusive development, approval and adoption of the data standards.