bluuebunny/crossref_metadata_2025_split huggingface dataset:
P008 Whole-genome RNA sequencing in collagenous colitis: a subgroup analysis
Authors: Escudero-Hernández C , van Beelen Granlund A , Bruland T , Sandvik A K , Koch S , Østvik A E , Münch A 
Published: January 2020

Abstract

Background
Collagenous colitis (CC) is a common inflammatory bowel disease that exhibits chronic watery diarrhoea. Treatment with the glucocorticoid budesonide is effective to induce and maintain clinical remission in most cases; however, few patients do not respond to treatment. To distinguish between different CC subgroups on a molecular level, we performed genome-wide RNA sequencing (RNAseq) analysis on mucosal samples.

Methods
We collected colonic biopsies from healthy controls (Hc) and CC patients with active disease. Additionally, we obtained matched samples from budesonide-responsive patients (clinical remission) under treatment with budesonide (9 mg/day, 8 weeks), and biopsies from budesonide-refractory patients (n = 9 patients/group). Ulcerative colitis (UC) samples were included as a separate control. Total RNA was isolated for RNAseq. Whole-genome expression data were analysed by principal component analysis (PCA) and differential gene expression (DGE) using R software. The Benjamini–Hochberg FDR method adjusted p values, considering &lt;0.05 as statistically significant. Gene-set enrichment analysis (GSEA) was performed using GSEA software and visualised in Cytoscape. Results were validated in tissue samples from the same patients by immunohistochemistry (IHC).

Results
PCA of all samples identified three principal components which separated sample groups into distinct clusters of gene expression, explaining 31% of the transcriptional variation. PCA clearly demarcated UC samples from Hc and CC. Mucosal samples from budesonide-refractory patients exhibited a discrete RNA expression profile that was distinct from all other groups. Moreover, PCA of budesonide-responsive persons also separated active from treated CC. Subsequent DGE analysis revealed differential expression of 395 genes in patients with active CC compared with Hc, and that these genes were mainly involved in immune response, cell cycle and apoptosis, according to GSEA. Using IHC, we confirmed the impaired proliferation and immune response on the protein level. In samples from budesonide-treated patients, 75 genes were differentially regulated compared with controls. A paired comparison of matched samples from CC patients before and after induction of remission showed an expression change of 337 genes, mainly involved in DNA recognition.

Conclusion
CC is a transcriptionally homogeneous disease that can be characterised by differential gene expression. Unique gene expression profiles describe patient sub-populations, which could define budesonide-refractory CC as a distinct disease entity. Further study of the transcriptional landscape of CC may reveal pathogenic mechanisms and therapeutic targets for this common, debilitating inflammatory bowel disease.
P013 The JAK-3 and TYK-2 / STAT pathways are activated in moderate to severe ulcerative colitis
Authors: Loza M , Brea J M , Calviño-Suarez C , Baston-Rey I , Ferreiro-Iglesias R , Dominguez-Munoz J E , Barreiro-de Acosta M 
Published: January 2020

Abstract

Background
Ulcerative colitis (UC) is a chronic, progressive and disabling disease with a complex pathology of unknown aetiology influenced by genetic, environmental and microbiota factors that lead to an immunological and inflammatory response in the colon. Janus Activated Kinase (JAK) family plays a key role in modulating the adaptive and innate inflammatory response. The JAK/STAT pathway involvement in UC has been demonstrated in both animal models and human studies. Thus, overexpressed JAK-3 has been detected in the intestine of patients with UC, suggesting a key role in their pathophysiology and the inhibition of TYK-2 in animal models resulted in an improvement of the disease, which would explain its implication in the inflammatory process. We hypothesise here that there could be an activation of JAK-3 and TYK-2 signalling pathways in UC patients. Thus, we aimed to detect the activation of both signalling pathways by means of western-blot studies in UC patient samples

Methods
A prospective, observational single-centre study was designed. Inclusion criteria were adult patients with endoscopic active UC (more than Mayo-0) confirmed in a programmed colonoscopy. All patients signed informed consent. Samples were obtained from overstock of routine biopsies in the more severe segment affected of the large bowel. Tissues were homogenised and processed in order to obtain cell lysates by employing RIPA buffer and ultrasounds. The degree of activation of the JAK-3 and TYK-2 pathways was measured by detecting the phosphorylation of both targets as well as of STAT1, STAT3, STAT4, STAT5 and STAT6 through western blot by employing specific antibodies for total and phosphorylated proteins.

Results
19 UC patients were consecutively included. Mean age was 46 years old. 53% were female, 47% were extensive colitis (E3) and 53% left-side colitis (E2). Regarding endoscopic activity, 26% had Mayo-1, 53% Mayo-2, and 21% Mayo-3. Immunoreactive bands for both phosphorylated JAK-3 and TYK-2 were detected in the biopsies from UC patients, evidencing that colonic inflammation leads to an activation of both targets. The study of STATs phosphorylation showed immunoreactive bands for phosphorylated forms of STAT1, STAT3, STAT4, STAT5 and STAT6 confirming the activation of both signalling-pathways in these patients (Figure 1).

Conclusion
The developed translational workflows involving basic/clinical research confirm the activation of both JAK-3 and TYK-2-dependent signalling pathways in UC patients, validating both kinases as targets for treating UC. The developed methodology allows studying the target engagement for future JAK-3/ TYK-2 inhibitors employed in clinical trials.
P016 Efficiency of evaluating the expression of Toll-like receptors 2, 4, 6 and proteins proteomic profiling as integral indicators for predicting the risk of early relapse of ulcerative colitis
Authors: Tarasova G , Volkov A , Iakovlev A , Ghazudin T , Shcherbakova I 
Published: January 2020

Abstract

Background
Success was made in the study of TLR in congenital and adaptive immunity, which determined a new look at immune processes at ulcerative colitis (UC). Modern achievements of proteomic methods of analysis research allow to define molecular characteristics of the inflammation in colon mucosa of patients with UC.

Methods
The study included 86 patients with UC, an average age of 39.0 ± 1.4 years. Groups: 1–15 (17.4%) patients with distal form of UC, 2–42 (48%) left-sided form, 3–29 (33.7%) patients with total UC. The expression of TLR on peripheral blood monocytes was determined in the immunofluorescence test. Two-colour analysis was performed on a flowthrough laser cytofluorimeter (Cytomics FC500, Beckman Coulte). The percentage of monocytes (CD14 + cells) carrying TLR2, TLR4, and TLR6 on their surface was assessed. The separation of proteins of colon mucosa was based on technologies of IEF, SDS–PAGE, 2DPAGE, by standard sets (MB-HIC C8 Kit, MB-IMAC Cu, MB-Wax Kit, Bruker, USA). The getting of mass spectrogram was determined by matrix-assisted laser desorption-ionisation time-of-flight mass spectrometry (MALDI-TOF-MS/MS, Ultraflex II, Bruker, USA). Statistical analysis was performed using the software Statistica 10.0 (Statsoft).

Results
The direct average relationship was established between the number of monocytes expressing CD14 + CD282 +, CD14 + CD284 +, CD14 + CD286 + and the area of inflammation (r = 0.49, r = 0.55, r = 0.42, p &lt; 0.05). The nonlinear regression equation was used. Calculation example: the risk of recurrence UC = exp (−26.1 + (0.4) × TLR 2)/(1 + exp (−26.1 + (0.4) × TLR 2)), χ2 = 130, 59, p &lt; 0.0001. Thus, when the number of monocytes expressing TLR2 is not more than 60%, the risk of recurrence of the UC is not more than 11%, with values above 70%, the probability of recurrence exceeds 80%. We identified potentially new molecular markers of the early relapse of ulcerative colitis: SMAD2 activates the transcription of TFG1β and leads to development of fibrosis in colon submucosa in patients with UC; significant decrease of the expression of PPARγ promotes the activation of STAT and AP-1 signalling pathways that promotes the increase of the synthesis of IL-2, 6, 8, 12, TNFα, the activity of immune and inflammation processes in colon mucosa; the reduction of expression of β-defensin-1 in cells of colon mucosa is accompanied with increased expression of CCR6 that promotes the formation of inflammatory infiltrates in colonic submucosa in UC.

Conclusion
Expression of TLR 2, 4, 6 in blood monocytes (the risk of recurrence UC ratio) and new protein molecular markers of colon mucosa (SMAD2, PPARγ, and apoС-III) can be used as a tool for prediction of early relapse UC.
P019 Colonic mucosal kinase activity, cytokine and chemokine profiles in inflammatory bowel disease
Authors: Brand E , Roosenboom B , Malvar Fernandez B , Lutter L , van Koolwijk E , van Lochem E G , Horjus Talabur Horje C S , Reedquist K A , van Wijk F , Oldenburg B 
Published: January 2020

Abstract

Background
With the approval of tofacitinib, an oral Janus Kinase (JAK) inhibitor, modulation of kinase activity has been added to the therapeutic armamentarium of inflammatory bowel disease (IBD). Despite its established efficacy, at least a third of patients will not respond to this or other therapeutic options such as anti-tumour necrosis factor (TNF), anti-interleukin (IL)23/IL12 compounds or vedolizumab. A better understanding of the inflammatory profile could aid in tailoring drugs to individual patients. We therefore explored mucosal cytokine, chemokine and kinase activity profiles in IBD.

Methods
Colonic mucosal biopsies were collected from (1) patients with Crohn’s disease (CD, N = 8), (2) patients with ulcerative colitis (UC, N = 8) and (3) healthy controls (N = 4). IBD samples were collected both from inflamed and non-inflamed tissue from the same patients. All IBD patients were biological-naïve and had not used corticosteroids in the past 3 months. Biopsies were snap frozen for later kinase activity determination or directly used in a 24-h explant culture. Whole biopsy kinase activity (tyrosine, serine and threonine kinases) was assessed using the Pamgene platform. A 64-analyte panel was examined in the supernatant of the cultured biopsies employing a multiplex assay (Luminex).

Results
Whole-biopsy kinase activity differed between inflamed and non-inflamed mucosa of IBD patients, with more overall tyrosine kinase activity in inflamed mucosa in UC, and serine/threonine kinase activity in inflamed mucosa in CD as compared with non-inflamed mucosa (Figure 1). The kinase activity profile of non-inflamed mucosa of CD and UC patients was similarly different from mucosa of healthy control participants (Figure 2). The cytokine and chemokine profile of inflamed biopsies differed from non-inflamed IBD biopsies and healthy control biopsies, with higher levels of S100A8, TNFα, IL-6, oncostatin M (OSM) and triggering receptor expressed on myeloid cells-1 (TREM-1), amongst others (Figure 3).

Conclusion
In IBD, inflammation in the mucosa can be characterised both by explant-culture and kinase activity assessment. The difference in kinase activity between non-inflamed IBD mucosa and healthy control mucosa suggests the presence of sub-clinical alterations in cell signalling. The observed differences in the kinase, cytokine and chemokine profiles underscore the importance of this approach in the elucidation of the pathophysiology in IBD.
P022 Impact of increasing disease symptom concordance between patients with inflammatory bowel disease and their physicians: insights from US, France, Germany, Italy, Spain and UK (5EU)
Authors: Kershaw J , Wild R , Lukanova R , Bruce Wirta S , Sikirica M 
Published: January 2020

Abstract

Background
Accurate disease activity assessment builds on dialogue between patients with CD and UC and their doctors and is key for management of IBD.

Objective
Assess the level of patient and physician symptom concordance on disease, health-related quality of life (HrQol), work productivity and patient satisfaction using the Adelphi Disease-Specific Programme.

Methods
Cross-sectional study among CD and UC patients and their gastroenterologists in US and 5EU. Data were collected in two waves, from November 2014–March 2015, and September 2017–January 2018, and then merged. Physicians completed patient record forms (PRFs) for the next seven UC and eight CD patients; same patients completed patient self-completion (PSC) forms describing how UC/CD affected them. PRFs collected diagnosis data, disease history, symptoms, treatment and management history. PSCs collected similar data, including patient reported outcome measures to assess the humanistic burden. Patients were grouped based on degree of alignment with physicians in symptom reporting (full agreement, discordance on 1–4 symptoms, discordance on 5+ symptoms out of 22 symptoms). Comparisons between groups were made using Chi-squared and Kruskal–Wallis for continuous and categorical variables.

Results
A total of 2524 CD and 2153 UC patients were included in the analysis (mean age CD 38.5/UC 40.3, CD 50.2% male/UC 52.0%, mean time since diagnosis CD 5.6 years/UC 5.1 years). There was overall reasonable agreement in symptom reporting between patients and their physicians; however, amongst 17% of UC and CD patients and physicians, there was discordance on 5+ symptoms. Higher symptom concordance significantly associated with better patient outcomes: higher remission rates, productivity, HrQoL scores and satisfaction (Table 1).

Conclusion
We observed moderate discordance between patient and physician reporting of current UC and CD symptoms. While even small discordance was associated with worse clinical outcomes: HrQol, productivity and satisfaction. Future studies are needed to improve understanding of the reasons behind discordance and association with worsened outcomes.
P027 Repertoire analysis of memory T-cell receptors in Japanese patients with inflammatory bowel disease
Authors: Takeru N , Kakuta Y , Izumiyama Y , Ichikawa R , Okamoto D , Moroi R , Kuroha M , Kanazawa Y , Kinouchi Y , Masamune A 
Published: January 2020

Abstract

Background
The T-cell receptor (TCR) repertoire was assessed in response to various antigens and was considered to be associated with the pathogenesis of inflammatory bowel disease (IBD). Thus, we performed TCR repertoire analysis to examine the pathology of IBD from changes in the TCR repertoire of memory T cells in the intestinal lamina propria mononuclear cells (LPMCs) and peripheral blood mononuclear cells (PBMCs) of patients with IBD.

Methods
LPMCs in the surgical specimens and PBMCs were isolated from 12 patients with IBD [five patients with ulcerative colitis (UC) and seven patients with Crohn’s disease (CD)]. PBMCs were collected from 10 healthy individuals as controls. Comprehensive TCR sequence analyses of adaptor-ligation PCR products were performed using MiSeq.

Results
The diversity of TCR-α and -β in PBMCs was significantly lower in patients with IBD than that in controls (p = 0.00084 and 0.0013, respectively). Comparisons of TCR diversity in LPMCs and PBMCs between CD and UC showed that the diversity in LPMC was not affected by diseases, whereas that in PBMCs was significantly lower in CD than in UC (p = 0.045 and 0.049, respectively). Some TCR clones may have shown a specific increase or decrease in CD and UC and many clones were common to both LPMCs and PBMCs in the same patients.

Conclusion
The diversity of TCR clones in LPMCs and PBMCs in patients with IBD was significantly lower than that of PBMCs in controls. TCR diversity in PBMCs was particularly low in patients with CD.
P028 The anti-inflammatory effects of a poly-probiotic on the oral mucosa
Authors: Marlina E , Hirani B , Mercadante V , Shephard M , Kishida S , Sebepos-Rogers G , McMillan R , Hodgson T , Leeson R , Porter S , Barber J , Fedele S , Smith A 
Published: January 2020

Abstract

Background
Probiotics have previously showed evidence of being efficacious in inducing and maintaining remission in post-operative recurrent pouchitis. The potential mechanism of action of probiotics has been attributed to their ability to reduce pro-inflammatory cytokine production, both within the mucosal tissue and systemically. We present our study which characterises the anti-inflammatory effects of probiotics on the oral epithelium and in the treatment of oral lichen planus (OLP), a chronic inflammatory disease of the oral mucosa.

Methods
VSL#3 (VSL#3-ACTIVE batch 703093 Exp date 04/2019) is a highly concentrated (450 billion live bacteria per sachet) poly-probiotic food supplement that contains eight different strains of live bacteria. The mouth ordinary epithelium cell line (MOE-1a) was stimulated with VSL#3 plus or minus the pro-inflammatory bacteria E. coli. The resultant effects on cytokine production and wound healing were measured using ELISA and live cell imaging. Wound closure was calculated using ImageJ software. OLP patients (n = 80) and healthy controls (n = 44) were recruited from UCLH Eastman Dental Hospital (Ethics 17/LO/0475) and saliva and blood samples tested for CXCL10 levels using an ELISA. OLP patients with active disease (n = 30) were recruited into a double-blind placebo-controlled proof-of-concept trial investigating the potential benefit of VSL#3 in the treatment of clinical symptoms (NCT03052179). Patients consumed two sachets twice daily for 30 days with a 30-day follow-up. Clinical questionnaires, saliva and peripheral blood were collected on days 0, 30 and 60. A daily quality of life and compliance diary was used by all participants.

Results
The addition of VSL#3 to MOE1a cells stimulated with E. coli resulted in a significant reduction in pro-inflammatory cytokine secretion and an acceleration in wound healing. OLP patients were found to have an elevation in the pro-inflammatory chemokine CXCL10 both locally (saliva) and systemically (serum) compared with healthy controls. Finally, the clinical trial demonstrated that VSL#3 was tolerated and safe for patients with OLP. Although there is no statistical evidence, descriptive results suggest that VSL#3 can confer some beneficial effects on patients with active OLP. We noted a reduction in the number of sites of disease activity and an improvement in quality of life in the VSL#3 group compared with placebo. Corticosteroid usage was also reduced in the VSL#3 group.

Conclusion
VSL#3 has the ability to improve oral epithelial wound healing and reduce pro-inflammatory cytokines secretion in vitro. In OLP, the consumption of VSL#3 seems to provide some clinical benefits, but due to the study size a more substantial multi-centre trial is necessary to confirm these observations.
P029 TRIM 21 protects against ulcerative colitis and colitic cancer via smad7 inhibition
Authors: Hahm K B 
Published: January 2020

Abstract

Background
Proteins of the tripartite motif-containing (TRIM) superfamily are critical in a variety of biological processes in either innate immunity or eliminating invading pathogens, by which had been implicated in pathogenesis of autoimmune diseases including inflammatory bowel diseases. The typical structure of TRIM proteins contains a RING motif in the N-terminal end, followed by a B-box motif, a coiled-coil domain and a B30.2/PRYSPRY region in the C-terminal end led to the regulation of TGF-β anti-inflammatory cytokines, by which TRIM21 has been reported to regulate IBD negatively through inhibiting Th1/Th17 cell differentiation.

Methods
Since antisense oligonucleotide targeting smad7 was withdrawn from clinical trial due to insufficient efficacy, in this study, we generated TRIM21 overexpressed cell lines to study the binding of TRIM21 to smad7 as well as the regulation of consequent TGF-β receptor.

Results
TRIM21 significantly binds to smad7 as well as repressed levels of TGF-b type I/II receptor. SBE-luc and 3TP-luc assay showed significantly decreased activities under TRIM21 + TGF-β. Since TRIM21 contains ubiquitin ligase, PRYSPRY, TRIM21 with TGF-β significantly decreased TGFRII via UPL. These in vitro evidences that TRIM21 significantly repressed TGF-β after binding smad7 were validated with DSS-induced colitis and colitic cancer model. TRIM21 was significantly decreased in DSS-induced ulcerative colitis, whereas ameliorated colitis showed significant restoration of TRIM21

Conclusion
Leading to conclusion that loss of TRIM21 led to significant bout of IBD.
P030 Dynamics of intestinal inflammation and microbial dysbiosis in pregnant women with IBD and their infants
Authors: Tarassishin L , Kim E , Eisele C , Barre A , Thjømøe A , Mørk E , Dubinsky M , Stone J , Nair N , Debebe A , Hawkins K , Rendon A , Sabino J , Torres J , Colombel J F , Peter I , Hu J 
Published: January 2020

Abstract

Background
IBD often affects women during their reproductive years; however, the effect of pregnancy on disease course remains poorly understood. We aimed to assess intestinal inflammation in IBD patients compared with controls as measured by faecal calprotectin (FC). We also investigated whether maternal IBD diagnosis was associated with altered FC in the offspring and if there were particular bacterial taxa that correlated with FC levels.

Methods
Pregnant women with or without IBD and their infants were prospectively enrolled in the MECONIUM study during 2015–2018 years. FC levels at each trimester of pregnancy and in babies throughout the first 3 years of life were measured using a quantitative enzyme immunoassay (CALPRO AS, Norway). Multivariate regression analysis was applied to investigate FC levels. Stool microbiota composition in the maternal and baby stool was assessed using 16s rRNA sequencing.

Results
617 faecal samples from 342 mothers (91 IBD, 251 control) and 1005 faecal samples from 288 infants (born to 76 mothers with and 212 without IBD) were analysed. FC levels in pregnant women with IBD were significantly higher than in control pregnant women regardless of IBD type (Crohn’s disease vs. ulcerative colitis). In IBD mothers FC levels showed a decline during pregnancy (µg/g, median (interquartile range); first trimester 130.8 (102.3–147.3) vs. third trimester 85.9 (44.5–125.4), p = 0.02) (Figure 1). Patients with flare at stool collection had the highest, while those treated with anti-TNF plus thiopurine showed the lowest FC at each trimester. Babies born to mothers with IBD presented higher FC levels than those born to control mothers at multiple time points between 2 and 36 months of age (Figure 2A); the median FC levels were the highest after 1 year up to 3 years in those babies whose mothers presented active disease during pregnancy (Figure 2B). Microbial α-diversity showed an inverse relationship with FC in both mothers (r = −0.25, p = 0.032) and babies (r = −0.31, p = 0.0035). Blautia and Streptococcus abundance were positively correlated with FC in babies.

Conclusion
FC levels in IBD patients decreased throughout pregnancy. Maternal IBD, lower microbiome diversity and the abundance of certain microbial genera were associated with higher FC in the offspring up to 3 years of life. These findings suggest a potential favourable impact of pregnancy on IBD activity and highlight a possible effect of IBD during pregnancy on the intestinal inflammation in offspring, which could be mediated through altered microbiome.