bluuebunny/crossref_metadata_2025_split huggingface dataset:
P007 Identification and characterisation of intestine-derived circulating resident memory T cells (ex-Trm) in health and Inflammatory Bowel Disease
Authors: Rodger B , Hoti I , Gordon H , Lindsay J , Stagg A 
Published: May 2021

Abstract

Background
Tissue resident memory T cells (Trm) persist in peripheral tissues where they protect against pathogens but can also contribute to inflammatory disease. Recent work shows that Trm can re-enter the circulation and give rise to new effector T cell and Trm populations in secondary tissue sites. Such ‘ex -Trm’ derived from the skin co-express the residency marker CD103 with cutaneous leukocyte antigen (CLA), a marker associated with skin tropism. Many T cells in the human intestine are Trm but it is unknown whether these cells re-enter the circulation; the existence of gut-derived ex-Trm would have important implications for IBD treatment targeting the recruitment of circulating gut-homing cells. Here, we identify a population of blood cells that co-express CD103 and the gut-homing integrin a4b7 and determine how they are changed in IBD.

Methods
Peripheral blood mononuclear cells (PBMCs) were isolated from healthy volunteers and patients with active IBD (Crohn’s disease or ulcerative colitis). Cell surface staining and multi-colour flow cytometry were used to identify CD4+ and CD8+ subsets of antigen experienced (CD45RA-) conventional T cells (abTCR+) and determine expression of markers associated with tissue tropism and residency.

Results
Staining with antibodies to CD103 and b7 integrin were used to define CD103b7+a4b7+ putative gut ex-Trm based on the excess per cell expression of b7 resulting from its contribution to both integrins. A separate CD103b7+a4b7- population defined by 1:1 expression of CD103 and b7 contained CLA+ skin ex-Trm. Gut ex-Trm comprised 0.3% total circulating CD8+ T cells (range 0.02–1.4%), and 1.2% CD4+ T cells (range 0.3–3%). Gut and skin ex-Trm were phenotypically similar; both expressed the residency associated markers CD101 and CD9 but lacked expression of CD69. Gut ex-Trm were phenotypically distinct from both traditional CD103-a4b7+ gut tropic CD45RA- antigen-experienced T cells and naïve T cells; significantly more gut ex-Trm expressed CD101 and CD9 and fewer expressed CD27. The proportion of gut ex-Trm did not differ between heath and IBD. However, the ratio of gut:skin ex Trm was significantly reduced in active Crohn’s disease but not ulcerative colitis indicating a selective reduction in the population derived from the intestine.

Conclusion
A putative population of gut-derived ex-Trm can be identified in the blood of healthy controls and IBD patients. This population has a distinctive phenotype similar to that of previously described skin-derived ex-Trm. Circulating ex-Trm could link discreet areas of intestinal inflammation in Crohn’s disease and there is a selective loss of the gut ex-Trm population from the blood of these patients. The role of ex-Trm in IBD merits further study.
P011 Serum adhesion G protein coupled receptor levels are associated with disease activity in pediatric patients with Crohn’s disease
Authors: Werner L , Weiss B , Shamir R , Shouval D 
Published: May 2021

Abstract

Background
Despite increasing number of therapies available for patients with Crohn’s disease (CD), response rates are generally not high. Therefore, there is a great need to identify novel targets in patients with active disease.

Methods
Serum samples were obtained from pediatric patients with CD at the time of diagnosis and following induction therapy with exclusive enteral nutrition (EEN) or anti-TNFa therapy, and from control subjects, defined as having normal colonoscopy without past or present history of inflammatory bowel diseases or other immune-mediated disorder. Associated clinical and laboratory data from these subjects was also collected. Serums were subjected to Olink technology (metabolism panel), based on proximity extension assay, enabling analysis of 92 proteins in each sample. Analysis was performed on a Normalized Protein eXpression (NPX) file comparing effect of different factors on the overall biomarker expression by supervised, multivariate, principal component analysis and verification by univariate ANOVA with Benjamini-Hochberg and post-hoc Tukey analysis.

Results
Eighty-eight serum samples were collected: 30 from control subjects and 58 from 32 patients with CD. The median age of patients in the control and CD groups was 13.9 years (IQR 11.1–16.6) and 14.6 years (IQR 12.2–16.9), respectively (P=0.32). Twenty-four patients were treated with anti-TNFa agents and 8 with EEN. The median pediatric Crohn’s Disease Activity Index decreased from 35 (22.5–42.5) to 5 (0–12.5, P&lt;0.001) following induction therapy. We identified 34 proteins that significantly differed between patients and controls. Moreover, within patients with CD, 8/34 of these proteins were differentially expressed in patients with active or non-active disease. Among these, adhesion G-protein coupled receptors E2 (ADGRE2) and G2 (ADGRG2), important for macrophage differentiation, were significantly lower among CD patients vs. controls. When stratifying according to disease activity, low levels of both markers were identified in CD patients with active disease (either patients that were treatment-naïve or those with primary non-response to anti-TNFa therapy) or with increased CRP levels. Principal component analysis showed that a combination of ADGRE2 and ADGRG2 showed good separation between control subjects and treatment naïve patients with active CD.

Conclusion
Serum levels of ADGRE2 and ADGRG2 are differentially regulated in patients with active CD, and should further be validated in additional Cohorts. The expression patterns and function of these receptors on different immune cells should be defined in future studies.
P019 Differential effects of first-line IFX and conventional treatment on inflammatory serum proteins of paediatric moderate- to-severe Crohn’s disease patients
Authors: Jongsma M , Costes L M M , Tindemans I , Cozijnsen M A , Raatgeep R H C , van Pieterson M , Li Y , Escher J C , de Ridder L , Samsom J N 
Published: May 2021

Abstract

Background
Recently, we established that First-line infliximab (FL-IFX) was superior to conventional treatment (CONV) in achieving and maintaining clinical remission in newly diagnosed paediatric patients with moderate-to-severe Crohn’s disease (CD) (PMID: 33384335). Selection of patients eligible for FL-IFX can be corroborated by combining clinical criteria with immunological biomarkers relating to the patients’ underlying immunopathology. Therefore, we investigated differential effects of FL-IFX and CONV induction treatment on serum inflammatory protein concentrations and assessed whether stratification of CD patients according to baseline serum profiles of IFX-modulated proteins relates to maintaining remission at week 52.

Methods
In the TISKIDS randomised controlled trial, newly diagnosed therapy-naïve CD patients with a weighted paediatric CD activity index (wPCDAI) &gt;40 were included. FL-IFX consisted of 5 IFX infusions (5 mg/kg) combined with azathioprine (AZA) maintenance treatment. CONV treatment consisted of induction therapy with Exclusive Enteral Nutrition or oral prednisolone combined with AZA maintenance. Serum samples of 48 FL-IFX and 43 CONV patients, collected at time of diagnosis and after 10–14 weeks of induction treatment, were available. Concentrations of 92 inflammatory proteins were determined with Olink Proteomics® proximity extension technology. Hierarchical clustering was performed to stratify patients according to their serum immune profiles. Clinical disease remission was defined as wPCDAI&lt;12.5.

Results
At baseline, prior to therapy, clinical disease characteristics and serum inflammatory protein concentrations did not significantly differ between FL-IFX and CONV treated patients. After 10–14 weeks, FL-IFX reduced 28 proteins and enhanced 2 proteins while CONV treatment reduced 13 proteins and enhanced 1 protein. Eighteen proteins were differentially regulated by FL-IFX only.
Hierarchical clustering of patients based on their baseline profiles of the 30 IFX-modulated proteins revealed 2 patient clusters. The CD-hi cluster had an increased abundance of 24/30 proteins amongst which Oncostatin-M, TNFSF14, HGF and TGF-alpha and higher clinical inflammatory parameters compared to the CD-lo cluster.
Crucially, both baseline inflammatory protein profile and type of induction therapy significantly determined whether patients reached clinical disease remission without treatment escalation at week 52; with CD-hi patients having the lowest change.

Conclusion
FL-IFX achieves stronger reduction and modulates more proteins than CONV treatment. Stratification of patients according to profiles of IFX-modulated proteins discerns patients with a lower chance of reaching clinical remission without treatment escalation at week 52.
P026 Microbial Diversity is affected by Cognitive Behavioral and Mindfulness-Based Stress Reduction and accompanied by an Inflammatory Response in Patients with Crohn’s Disease
Authors: Ilan K , Motro Y , Nemirovsky A , Lerner L , Schwartz D , Goren G , Sergienko R , Greenberg D , Slonim-Nevo V , Sarid O , Friger M , Regev S , Odes S , Moran-Gilad J , Monsonego A 
Published: May 2021

Abstract

Background
Crohn’s disease (CD) is associated with psychological stress and alteration of gut microbiota as compared to healthy individuals. Previous work has reported imbalances in CD patients across four major bacterial phyla including Firmicutes, Bacteroidetes, Proteobacteria and Actinobacteria. We aimed to determine the effect on gut microbiota of a 3-month period of Cognitive Behavioral and Mindfulness-Based Stress Reduction (COBMINDEX), a psychological intervention shown to improve the wellbeing and inflammatory state of CD patients.

Methods
Microbial analysis of stool samples, circulating inflammatory markers and a wide range of psychological parameters related to stress, well-being and daily activities were measured and compared among 25 CD patients before (T1) and after (T2) COBMINDEX, and 25 matched CD wait-list controls at the corresponding time-points T1 and T2. Stool DNA extracts were subjected to 16S amplicon sequencing (Illumina) and analysed for taxonomical abundance, alpha and beta diversity using the QIIME2 pipeline and correlated with inflammatory and clinical parameters.

Results
Microbial alpha diversity among all CD patients at T1 significantly correlated to the 3 key cytokines: IL-10 (p=0.04, correlation=0.48), INFg (p=0.02, correlation=0.53), and INFa (p=0.02, correlation=0.51). At T2, while beta diversity was significantly increased in the COBMINDEX group (p=0.03), it was significantly decreased (p=0.0001) among the wait-list controls (Figure 1). Furthermore, compared with the wait-list controls, changes in inflammatory and clinical markers such as INFg, TNFa, IL-6, calprotectin and CRP, occurring following COBMINDEX, were accompanied by changes in the 4 main phyla including Firmicutes, Bacteroidetes, Proteobacteria and Actinobacteria (Figure 2). Lastly, changes in a variety of unique taxa, not previously implicated in CD, were also associated with changes in inflammatory and clinical markers such as INFg, TNFa, IL-6, calprotectin and CRP in the COBMINDEX but not in wait-list controls (Figure 2).

Conclusion
Our results show that microbial diversity is connected to the inflammatory profile of CD patients, and is significantly altered by COBMINDEX. Moreover, changes in the 4 main phyla that are known to be different among CD patients are linked to changes in various inflammatory markers, demonstrating the microbial-inflammatory relationship among CD patients. Lastly, we found that changes in the abundancies of 7 taxa, hitherto untied to CD in the literature, correlate with inflammatory markers, hinting at new microbial targets in CD.
P044 Transcriptome analysis identifies dysregulated pathways and targets for therapy in patients undergoing surgical resection for Crohn’s disease
Authors: Akiror S , Acharjee A , Jeffery L , Shivaji U N , Zardo D , Gkoutos G , Ghosh S , Iacucci M 
Published: May 2021

Abstract

Background
In Crohn’s disease, intestinal strictures develop in 40% of patients often requiring repeated surgeries. Current treatments have limited efficacy. Therefore, better understanding of dysregulated molecular pathways is needed to identify targets for therapy. The aim of this study was to identify novel pathways involved in stricture pathogenesis.

Methods
Patients undergoing resection for CD-related strictures were recruited (IRAS ID-248494). RNA was extracted from proximal, strictured and distal segments of resected ileal/ileo.cecal segments. cDNA libraries were prepared for 9 patients using QIAseq UPX 3’ Transcriptome reagents and sequenced. Normalised expressions were obtained through the CLC-Genomics Workbench (Qiagen). Differentially expressed genes (adj.P.Val&lt;0.05) were determined using the Limma package and PCA and PLS-DA modelling performed. Pathway-related genes were enriched using EnrichR. Area under the curve (AUC) analysis was done by the Random Forest method and qPCR used for gene validation.

Results
Strictured, proximal and distal tissues had unique transcriptomes; stricture segment clustered separately from proximal and distal segments, which were very similar by comparison. 64 genes were commonly up-regulated and 17 down-regulated in stricture compared to either control segments. Pathway enrichment on these 81 genes identified 30 targets. 3 of 5 down-regulated genes, associated with the epithelial barrier (KLF5, KRKT20, DSK) whilst most up-regulated genes related to extracellular matrix and tissue remodelling (COL1A1, CTSK, HYAL2, GREM-1, SERPINE1), inflammation (PECAM1, CCL2, EDNRB, LY96, CTSK), adipogenesis (IGF-1, NDN, HADHA) and cellular stress.
Based on statistical significance and biological relevance LY96, AKAP11, SRM, GREM1, EHD2, SERPINE1, HDAC1 and FGF2 were further studied for association with stricture. A combined AUC score of 0.979 (95%CI 0.902–1) confirmed strong association and was supported by qPCR validation with additional patients. (AUC=0.81, (0.56–0.95)).

Conclusion
Loss of epithelial integrity, increased inflammation, tissue remodelling and adiposity characterise the stricture and may be modulated by existing anti-fibrotic drugs not tested in Crohn’s disease as well as new drug development.
P076 The endocannabinoid system in the intestinal biopsies of children with Inflammatory Bowel Disease
Authors: Creoli M , Strisciuglio C , Vitale A , Oglio F , Paino S , Luongo L , Tortora C , Argenziano M , Di Paola A , Maione S , Rossi F 
Published: May 2021

Abstract

Background
The endocannabinoid (EC) system has been recently indicated as a possible therapeutic target in inflammatory bowel disease patients (IBD). G-protein-coupled cannabinoid receptors type 1 and 2 (CB1 and CB2) are the two most important components of the EC system. In our previous study, we demonstrated that a specific CB2 variant contributes to the risk for pediatric IBD, especially UC, and that this variant is associated with a more severe phenotype in both UC and CD. The aim of this study was to directly evaluate on intestinal biopsies of pediatric IBD patients the expression of EC system receptors and of several molecules associated with inflammatory pathway in IBD.

Methods
In this preliminary study, we analyzed the CB1 and CB2 expression in the intestinal mucosa of paediatric patients affected by Crohn’s disease (CD) or ulcerative colitis (UC), recruited at diagnosis, and of non-IBD healthy controls (HC). Three patients were enrolled for each group. In particular, we analyzed biopsies taken from the ileum and rectum, which are the tract often more inflamed in CD and UC respectively. Moreover, we evaluated Toll-like receptor 4 (TLR 4), interleukin- 6 (IL6), interleukin-1 β (IL1-β), prokineticin receptor 2 (PKR2) expression, which are known as pro-inflammatory elements. The expression of these receptors and pro- inflammatory cytokines was analyzed by western blot and immunofluorescence.

Results
We found an increased expression of CB2 and TLR4 in UC rectum biopsies compared to CD and HC children. Accordingly, there was also a significant increase of IL-6 expression in UC rectum biopsies compared to CD and HC children (p≤0.05 UC vs HC), as well as, we identified a high expression of IL-6 in CD ileum compared to UC and HC patients. Moreover, in CD ileum, we found an increase in TLR4 compared to UC and HC ileum. We did not find statistical difference for the low number of analyzed samples, but to be a preliminary work we are confident to reach statistical significance increasing the samples enrolled.

Conclusion
In according with our previous data, our results confirm that the EC system and in particular the receptor CB2 is more expressed in the intestine of IBD patients, especially in the rectum of UC patients, which is the tract with greater inflammatory involvement. We can speculate that CB2 receptor is involved in the pathogenesis of paediatric IBD, but additional functional studies are required to understand how the EC system contributes to disease development in pediatric IBD, and if these cannabinoid receptors might represent a new disease marker and could be used as a potential therapeutic target.

Expression of CB2 receptor in rectum biopsies
P077 What matters to Ulcerative Colitis patients when they make treatment decisions? A systematic review
Authors: Folan A M , Jones G , Baker D , Brown S , Lee M , Lobo A 
Published: May 2021

Abstract

Background
The decision for ulcerative colitis (UC) patients to opt for elective surgery or continue medical treatment is dependent on patient preferences taking into account a range of factors. In addition to choosing between medical and surgical treatment, patients undergoing elective surgery are presented with a further decision regarding which operation to choose. The aim of this systematic review is to identify and understand what matters to UC patients when they are making these decisions.

Methods
Five electronic databases (PubMed, Scopus, CINAHL, Medline, and Embase) were searched for relevant literature up to 15 October 2020. Qualitative, quantitative and mixed-methods studies were included in this review. Studies reporting on what was important to UC patients (over 16 years of age) when they make treatment decisions were included. The Mixed Methods Appraisal Tool was used to assess the quality of the papers. Thematic analysis was used to analyse the data.

Results
The searches identified 6,917 papers and a final 19 (eight quantitative, seven qualitative, four mixed methods) papers were included. All studies were published since 2007 and included a total of 3,328 participants from nine countries. Five overarching themes (and their associated 20 sub-themes) were generated to describe the factors reported as important to UC patients in making treatment decisions. These were: 1. Information provision (information content, knowledge about their illness, quality of information); 2. Impact of the treatment upon daily life (controlling physical symptoms, quality of life); 3. Levels of risk (trade-off, high risk, concern and worry); 4. Burden of treatment (the need to see benefits of medication, route and size of medication, side effects, dosing frequency, costs, effort of being the patient, adherence to medication, surgery concerns, timing of surgery); and 5. Patient-clinician relationship (shared decision-making, communication, mismatch between what clinicians and patients consider to be important).

Conclusion
Communication between patients and their IBD teams should take into account the range of factors that influence their treatment decision making. Decision support interventions that incorporate such factors may better support the patient-clinician relationship and improve knowledge of treatment options and how these impact on what matters to them. Future studies are needed to determine which factors identified in this review are dominant.
P079 Valproic acid increases MiR449a expression in intestinal mucosa from patients with inflammatory bowel disease
Authors: Felice C , Lewis A , Armuzzi A , Lindsay J O , Silver A 
Published: May 2021

Abstract

Background
Histone acetylation/deacetylation and microRNAs (MiRNA) are epigenetic mechanisms involved in the pathogenesis of inflammatory bowel diseases (IBD). Histone deacetylase (HDAC) inhibitors, such as valproic acid (VPA), demonstrated anti-inflammatory and anti-tumour effects in animal models of colitis.1,2 Modulation of MiRNAs is hypothesized to be a possible mechanism of action of HDAC inhibition.3 Recently, VPA was shown to modulate histone-3 (H3) acetylation and cytokine production in the human gut.4 In this study, we analysed whether VPA influences MiRNA expression in ex-vivo biopsies from IBD patients.

Methods
Colonic biopsies from IBD patients collected during routine endoscopic procedures were cultured with VPA (5 mM) or control media for 24 hours. Total RNA was extracted and a MiRNA array was performed (miScript™ miRNA PCR Array), profiling the expression of 84 MiRNAs predicted to regulate pro- or anti-inflammatory genes. Significantly altered MiRNAs were then validated in an independent cohort of samples using quantitative (q) RT-PCR. Caco-2 (ATCC® HTB-37™) cell cultures treated with VPA (5 mM) or control media were used to confirm changes of H3 acetylation and MiRNA expression. After 24 hours, H3 acetylation was evaluated by immunofluorescence and apoptotic markers were analysed by qPCR.

Results
Colonic biopsies from inflamed mucosa of 6 IBD patients (2 with Crohn’s disease (CD) and 4 with ulcerative colitis (UC)) were cultured with VPA or control media. MiRNA analysis showed a significant increase of miR449a, miR373-3p, miR300 and miR520d-3p in treated biopsies compared to control (p=0.004, p=0.024, p=0.04 and p=0.014, respectively). Validation was performed in an independent cohort of 14 IBD patients (7 CD; 7 UC), confirming a significant increase of MiR449a (p=0.026) in VPA treated biopsies. VPA treated Caco-2 cell cultures showed a significant increase of H3 acetylation density level (p=0.02), confirming the HDAC inhibitory activity. Gene expression analysis showed increased Caspase-3 (p=0.015), a pro-apoptotic marker, and decreased BCL-3 (p&lt;0.01), an anti-apoptotic marker. The level of MiR449a expression in VPA treated cells was significantly elevated (p&lt;0.01).

Conclusion
VPA increases the expression of MiR449a in cultured ex-vivo biopsies and in Caco-2 cells. This MiRNA has recently demonstrated to be involved in IBD pathogenesis and in colitis-associated colon cancer. This may represent a new insight into IBD pathogenesis and colitis-associated colon cancer development. Further work is required to determine the functional significance of miR-499a in the human intestine.

References