bluuebunny/crossref_metadata_2025 huggingface dataset:
DOP68 Thiomyristoyl ameliorates colitis by blocking the differentiation of Th17 cells via STAT3/IL-6 signal pathway
Authors: XU Y , zhang M , Xu G , Zou X
Published: January 2020

Background

The pathogenesis of inflammatory bowel disease (IBD), including Crohn’s disease (CD) and ulcerative colitis (UC), has not been fully elucidated, but a strong correlation between IBD and the immune dysregulation of the host has been persistently detected. Sirtuin 2 (SIRT2), a histone deacetylase, has recently been found to play an important role in inflammation, infection and immunomodulatory. As a potent SIRT2-specific inhibitor, thiomyristoyl (TM) has an extensive anticancer activity, but its anti-inflammatory property remains unclear.

Methods

Human peripheral blood mononuclear cells (PBMC) were differentiated into T helper 17 (Th17) cells in vitro and were treated with TM simultaneously. Interleukin (IL)-17A in the culture supernatant, the ratio of T cells, the levels of phosphorylated-signal transducer and activator of transcription (p-stat3) and phosphorylated-nuclear factor kappa-B (p-NF κB) were determined by enzyme-linked immunosorbent assay (ELISA), flow cytometry or western blotting, respectively. Then, colitis C57BL/6J mice induced by 2.5% dextran sulphate sodium salt (DSS) were treated with TM, and were evaluated by disease activity index (DAI) and haematoxylin-eosin (HE) staining. T-cell subsets in the mice spleen, IL-6, IL-10 and IL-17A in serum, related factors retinoic acid receptor-related orphan receptor-γ t (ROR-γt), forkhead box protein P3 (FOXP3), IL-17A, interferon γ γ (IFN-γ), IL-23, IL-10 and hypoxia-inducible factor (HIF)-1α in mice colon were measured by flow cytometry, ELISA, quantitative real-time polymerase chain reaction (Q-PCR) and western blotting, respectively.

Results

Compared with the positive control group, the levels of IL-17A in culture supernatant, the percentage of Th17, the levels of p-stat3 and p-NF kappa B in cell lysate were lower in the TM group. Compared with PBS-treated colitis mice, TM-treated colitis mice had longer colons, fewer weight-losses, lower DAI and histopathologic scores. Interestingly, although the expression of IFN-γ, IL-17A, and ROR-γt was inhibited in the colons of TM-treated mice, the level of FOXP3 did not change. Consistently, the percentage of spleen Th17 cells was decreased in the TM group while the percentage of Treg cells was not affected. In addition, the TM group had reduced levels of IL-23 and HIF-1α, and an increased level of IL-10 in the colon, compared with colitis group.

Conclusion

TM ameliorates DSS-induced experimental colitis by blocking the differentiation of Th17 cells, which may be associated with the STAT3/IL-6 signal pathway. SIRT2 may represent a potential target for the treatment of IBD.
DOP76 Corticosteroid sparing effects of ustekinumab therapy for ulcerative colitis through 2 years: UNIFI long-term extension
Authors: Peyrin-Biroulet L , Sandborn W , Sands B , Scherl E , Marano C , O’Brien C , Zhang H , Johanns J , Zhou Y , Abreu M , Arasaradnam R P , Rowbotham D S , Leong R W L , Danese S
Published: January 2020

Background

Ustekinumab (UST) is an IL12/23 blocker approved for Crohn’s disease and ulcerative colitis (UC). In the UNIFI maintenance study of patients with moderate–severe UC, >90% of the patients who achieved clinical response or remission at week 44 were able to eliminate corticosteroids, an important goal of therapy. In this analysis, we describe the corticosteroid (CS) sparing effects of UST treatment through week 92 among patients who were treated in the UNIFI long-term extension (LTE).

Methods

Responders to UST 8 weeks after IV induction entered the maintenance study and were randomised to UST 90mg SC (q12wks or q8wks) or PBO. Patients who did not respond to UST IV induction received a dose of UST 90mg SC at week 8 and were evaluated at week 16. Those in clinical response at week 16 (delayed responders), were not randomised on entry into the maintenance study and received UST 90mg q8wk. All patients who completed week 44 of the maintenance study were eligible to enter the LTE and continue to receive the same dose. Patients could receive UST dose adjustment during the LTE (q12wk to q8wk or q8wk to q8wk [sham dose adjustment]) from week 56 onward. Patients who remained on PBO were discontinued from the LTE after maintenance study unblinding. During the maintenance study, CS tapering was recommended for all patients receiving CS at maintenance baseline. At week 92 of the LTE, CS-free symptomatic remission rates and CS use were calculated using intention-to-treat analyses with a dose adjustment considered to be a treatment failure and with a dose adjustment not considered to be a treatment failure.

Results

Of the 284 patients in the randomised population who were treated with UST in the LTE, 139 were receiving CS at maintenance baseline. Of these, 92.8% (n = 129) were CS-free at week 92. CS-free symptomatic remission rates at week 92 are summarised in Table 1. Results were similar for the q8wk and q12wk maintenance doses. Of the UST-treated patients who were in symptomatic remission at week 92, 98.4% (182/185) were CS-free; similar results were seen when dose adjustment was not considered to be a treatment failure. Of the 116 delayed responders (non-randomised population) who continued to receive UST in the LTE, 92 patients were in symptomatic remission at week 92; of whom, 94.6% (87/92) were CS-free.

Conclusion

UST maintenance therapy, with both q8wk and q12wk dosing regimens, was effective in reducing and eliminating the use of CS in patients with UC. Nearly all patients in the LTE who were in symptomatic remission at week 92, were not taking steroids.
DOP88 Visceral fat area correlates well with anti-TNFα drug levels and secondary loss of response in Crohn’s disease patients
Authors: Lim Z , Welman C , Raymond W , Thin L
Published: January 2020

Background

A high body mass index (BMI) is known to adversely affect anti-TNFα trough levels and secondary loss of response (SLOR); however, the literature is scarce in defining what aspect of body mass determine these outcomes. We hypothesise that a large visceral fat area is associated with a lower anti-TNFα level and a higher rate of SLOR. Our aim was to determine the impact of fat and muscle compartment areas on these outcomes.

Methods

Crohn’s disease (CD) patients who were prescribed standard doses of an anti-TNFα agent [5 mg/kg, 8 weekly infliximab (IFX) or 40 mg EOW of adalimumab (ADA)] from 1 February 2015 to 30 June 2018 were examined retrospectively. Primary responders with a minimum therapy duration of 12 weeks, at least 12 months of follow-up and a trough level within 6 months of a CT or MRI, were eligible for inclusion. The primary outcome was the trough level and the secondary outcome was time to SLOR, defined as a need to dose escalate, change out of class, requiring ≥3 courses of corticosteroids in a 12-month period, or surgery. Patients were followed until they met a SLOR or the census date of 30 June 2019. Visceral fat area (VFA), subcutaneous fat area (SFA) and skeletal muscle area (SMA) were measured on the CT/MRI at the L3 vertebral level in cm2 by a radiologist blinded to the clinical outcome and corrected for patient height (cm2/m2).

Results

Of 813 patients prescribed an anti-TNFα agent in the study period, data from 69 eligible CD patients were included for analysis. The median age was 43.5 ± 16.2 years, and 42 (60.9%) were males. Forty-four (63.8%) and 25 (36.2%) patients were treated with IFX and ADA respectively. The mean BMI was 26.9 ± 5.2. Univariate analysis of infliximab trough levels found that total fat area, VFA, visceral fat index [VFI (VFA/height in m2)] and VFA/SMA ratio were inversely correlated with anti-TNFα trough level (p < 0.05). No association was found between ADA trough level and muscle/fat areas. After multivariate adjustment for CRP, albumin, presence of antibodies, concurrent immunomodulator use and gender, IFX levels were inversely associated with VFA [-0.021 (−0.038, −0.003) p = 0.025], VFI [−0.066 (−0.119, −0.013) p = 0.016] and VFA/SMA [−3.805 (−7.132, −0.477) p = 0.026] but not BMI [−0.232 (−0.520, 0.055) p = 0.11). No association was found for ADA trough levels. Kaplan–Meir analyses showed a trend towards a shorter time to SLOR with an increasing tertile of VFI in both IFX and ADA treated patients.

Conclusion

Visceral fat area corrected for height (VFI) is a better determinant of anti-TNFα trough levels and SLOR than BMI.
P001 The novel DYRK1a inhibitor VRN024219 alleviates disease severity on the IBD mouse models by modulating T-cell differentiation
Authors: Seo D H , Ma H W , Kim S , Kim D H , Kim H K , Lee S H , Kim S , Ahn J Y , Yoon S H , Kim D K , Cheon J H
Published: January 2020

Background

DYRK1A belongs to dual-specificity tyrosine (Y) phosphorylation regulated kinase (DYRK) family which is known to be activated through autophosphorylation of tyrosine residues in the activation loop and phosphorylates their substrates on serine and threonine residues. Other members of this family include DYRK1B, DYRK2, DYRK3, and DYRK4. Studies have revealed that DYRK kinase family plays an important role in regulating cell proliferation and apoptosis. DYRK1A has been reported to be strongly expressed in the brain and known to regulate various functions. However, the role and underlying mechanisms of DYRK1a in inflammation in general, specifically in IBD, remain poorly understood. Accordingly, we present the underlying mechanisms of DYRK1a on the course of IBD by using the novel DYRK1a inhibitor VRN024219.

Methods

First, we tested the effects of our compound VRN024219 on T-cell differentiation using naïve CD4 T cells extracted from the mouse spleen. Then we assessed the efficacy and mechanism of VRN024219 on the dextran sodium sulphate (DSS) and T-cell transfer-induced experimental colitis that mimics human ulcerative colitis (UC), comparing to that of Tofacitinib. Finally, we evaluated the effect of VRN024219 on pro-inflammatory cytokines such as interleukin (IL) −17A, IL-6, and tumour necrosis factor (TNF) α expressed by peripheral blood mononuclear cells (PBMCs) from 20 UC patient samples (Severance Hospital, Seoul, Korea).

Results

When VRN024219 was treated to splenocyte, the compound significantly downregulated Th17 and enhanced T reg cell differentiation. Protein levels of IL-17a, IL-6, and TNF α were increased in the DSS-induced colitis mice, whereas administration of DYRK1a inhibitor VRN024219 substantially improved clinical score than that of Tofacitinib-treated group. Additionally from T-cell transfer-induced colitis model, VRN024219 treated group demonstrated a larger population of ROR γ T-cell than that of tofacitinib Finally, the protein levels of proinflammatory cytokines were significantly down-regulated in VRN024219 treated 20 patient samples.

Conclusion

Our data provide clear evidence that the novel DYRK1a inhibitor plays a protective role in DSS- and T-cell transfer-induced colitis which was closely related to a Th17/Treg modulation. Thus, VRN024219 might a promising candidate for a new drug for IBD.
P020 MAM, an anti-inflammatory protein derived from Faecalibacterium prausnitzii as a biomarker in Crohn’s disease?
Authors: McLellan P , Lavelle A , Brot L , Straube M , de Sordi L , Grill J P , Bourrier A , Kirchgesner J , Chatel J M , Beaugerie L , Sokol H , Seksik P
Published: January 2020

Background

The intestinal microbiota of patients with Crohn’s disease (CD) is characterised by a specific dysbiosis, including the loss of Faecalibacterium prausnitzii (F. prau), a major bacterium with anti-inflammatory properties. MAM protein (microbiotia anti-inflammatory molecule) produced by F. prau has recently been identified as partially carrying the anti-inflammatory activity of this bacterium. The description of microbiota for diagnostic purposes in CD is currently difficult to achieve. MAM could be a biomarker of dysbiosis. Our goal was to study the expression of MAM in the intestinal ecosystem and to evaluate its interest as a biomarker.

Methods

An in silico approach was used to determine primers of MAM gene sequences specific to two currently known phylogroups (I and II) of F. prau. Faecal samples were collected from a monocentric cohort of CD patients in flare (n = 24) and in remission (n = 24) and from healthy controls (n = 12). To measure MAM expression in vivo in the microbiota, we extracted total RNA and DNA from each sample and performed quantitative PCR (qPCR) with MAM primers (I and II), F. prau primers and universal bacterial primers. We compared the relative expression (ΔCt) of MAM mRNA and DNA with qPCR to the overall amount of bacteria in faecal samples.

Results

MAM analyses of F. prau phylogroup II were the most informative. We observed that the relative expression of MAM mRNA was statistically lower in active patients 5.84 × 10−5 [95% CI 3.8 × 10−6–2.1 × 10−3] and in remission 5.36 × 10−5 [95% CI 2.5 × 10−8–1.9 × 10−2] compared with healthy controls 5.42 × 10−4 [95% CI 8.5 × 10−5–9.1 × 10−2], (p = 0.004). There was no difference in ΔCt between flare and remission in CD patients. The most relevant relative expression threshold was 1.6 × 10−4. At this threshold, sensitivity was 81%, specificity 76%, positive predictive value 94% and negative predictive value 47%. In multivariate analysis, the relative expression of MAM was an independently associated with the diagnosis of CD; OR 2.961 [95% CI 2.026–4.831] (p = 0.003). Using relative expression of MAM DNA, we found a significant decrease in the flare group compared with remission patients and controls. We found a negative correlation of MAM DNA expression with CRP (p = 0.0053) and HBI (p = 0.025).

Conclusion

Our results showed that the loss of MAM RNA expression is observed in CD patients. Moreover, MAM DNA detection was associated with CD activity parameters. This opens up the prospect of an ecological biomarker of CD based on the expression of molecules from the intestinal ecosystem. These results will need to be validated in larger independent cohorts and their specificities compared with other pathological situations.
P021 The influence of short-chain fatty acids produced by commensal bacteria on macrophage phenotype
Authors: Jukes C , Ricci L , von Coburg E , Nasr M , Walker A W , Duncan S H , Hegazy A N
Published: January 2020

Background

Macrophages are dynamic immune cells that react to environmental signals to adapt to their environment. Among this diverse array of signals are multiple bacterially derived metabolites including short-chain fatty acids (SCFAs) in the gastrointestinal tract. It has been previously shown that butyrate, one of several SCFAs produced by bacteria, is able to drive an antimicrobial phenotype in macrophages. However, it is still unknown which bacterial species are able to drive this impact and whether this effect can be recapitulated in vivo. The identification of a common molecular signature that drives this phenotype may allow us to easily identify bacteria with this beneficial phenotype and give insight into potential therapeutic strains.

Methods

Monocytes were differentiated into macrophages in the presence of SFCAs or bacterial supernatants from a wide range of gut commensals isolated from healthy human donors. RNA was isolated post-differentiation and gene expression screened using a defined gene panel using the Fluidgm Biomark system. In addition, cytokines present in the supernatant were measured by ELISA and the ability of the macrophages to kill intracellular bacteria determined. Currently, we are carrying out the same procedure using bone marrow-derived macrophages (BMDM) from mice in order to expand this work flow into the mouse system so that we can determine if bacteria identified can elicit their impact in vivo.

Results

We have screened numerous SCFAs alone and in combination in the bacterial killing assay to determine how these are able to influence macrophage phenotype. From these experiments, we have repeated the earlier observation that butyrate can increase macrophage killing of intracellular bacteria. In addition, we have identified four bacterial isolates that induce significantly increased microbial killing in macrophages. This is associated with the production of butyrate, acetate and formate. Using RNA from these different conditions, we are in the process of determining if there is a common molecular pathway by which this increased antimicrobial phenotype is driven.

Conclusion

Both SCFAs and SCFA producing bacteria are able to induce an antimicrobial phenotype in macrophages that increased the ability of these cells to kill intracellular pathogens. Identification of a common signature that drives these changes could help to screen bacterial isolates for this beneficial trait. Ongoing work hopes to identify such a signature, to enable us to screen large numbers of commensals using high-throughput molecular methods. This can then be verified using both in vitro and in vivo techniques.
P055 Effect of Mimosa caesalpiniifolia extract on DSS-induced colitis in Wistar rats
Authors: Garnevi Fávero A , Cordeiro T S , Silva K N , Cardili L , Silva M J D , Paiotti A P R
Published: January 2020

Background

Iinflammatory bowel disease (IBD) is a chronic disease characterised by dysregulation of the immune function response and imbalanced release of cytokines and unresolved inflammatory progress associated with the intestinal mucosa. In this way, new drugs are constantly tested as a novel treatment option. Mimosa caesalpiniifolia (MC) has been described as a potent analgesic, anti-inflammatory, anti-haemorrhagic, astringent, anthelmintic and diuretic. This study aimed to evaluate the effect of MC extract on dextran sulphate sodium (DSS)-induced colitis in Wistar rats.

Methods

Forty male Wistar rats were randomised into five groups: Sham; 5% DSS-induced colitis; control MC extract (125 mg/kg/day); 5% DSS-induced colitis treated with MC (125 mg/kg/day); 5% DSS-induced colitis treated with MC + 5-ASA (125 mg/kg/day for both). Colitis was induced by administration of 5% DSS (MP Biomedicals), diluted in drinking water and offered ad libitum for 5 days. MC plant extract was diluted in water and administered by oral gavage per 6 days. The leaf extracts were prepared in 70 gl ethanol and dried in a Buchi B19 Mini-spray dryer using 20% aerosol. The selection of the operating conditions used was followed by the Laboratory of Medicinal and Phytotherapeutic Plants of Unifal-MG. After the induction of colitis, body weight, general health status and faecal characteristics will be evaluated daily. All rats were euthanised on the ninth day of the experiment. The entire colon was dissected and fixed in 10% neutral-buffered formalin at room temperature and embedded in paraffin to provide sections for histological evaluation (Ethics Committee on the Use of Animals number 4362180119). The severity of colitis was evaluated in sections stained with Hematoxylin and Eosin. Parameters such as inflammation extent, regeneration and crypt damage were graded according to Dieleman et al. Statistical analysis was performed by one-way analysis of variance (ANOVA), followed by Tukey post hoc test using Graph Pad Prism (version 6.0).

Results

MC extract treatment reduced significantly the severity of DSS-induced colitis evidenced by a decreased in clinical symptoms (p < 0.0001). The rats treated with MC extract showed a decreased in diarrhoea episodes and rectal bleeding. We observed histopathological changes of colon tissue between the DSS-treatment and control groups, however, without statistical significance. The MC extract treatment reduced the damage to the colonic tissue evidenced by a decreased of inflammatory cells, ulcerations, and goblet cell preservation.

Conclusion

MC extract reduces the clinical signs and suggests a potential anti-inflammatory effect on DSS-induced colitis.
P061 The molecular landscape of perianal fistula in Crohn’s disease: opportunities for new therapeutic approaches
Authors: Bislenghi G , Verstockt B , Sabino J , Caenepeel C , Verstockt S , Wolthuis A , Ferrante M , D’Hoore A , Vermeire S
Published: January 2020

Background

Perianal Crohn’s disease (CD) is a debilitating condition, often refractory to medical therapy and requiring repetitive surgical interventions. Nonetheless, its pathophysiology is very poorly understood. Hence, we molecularly characterised the fistula tract and compared it to the molecular landscape of its inner rectal orifice.

Methods

We collected paired surgical biopsies from the fistula tract and the inner rectal fistula orifice in 29 CD patients with draining perianal fistula, requiring surgical examination under anaesthesia. RNA was extracted and single-end RNA sequencing performed using Illumina HiSeq4000. Sequencing data were analysed through differential gene expression (DESeq2) and corrected for the presence of proctitis. A false discovery rate of 0.001 was considered significant. In addition, cellular deconvolution methods (CIBERSORT) were applied to study the cellular composition of the fistula tract.

Results

Differential gene expression revealed 2701 transcripts being differentially expressed (1727 up, 974 down in fistula) between the fistula tract and the paired rectal mucosa. The top upregulated gene, LBP (fold change [FC]=2858.8, p = 3.6E−13), highlights the potential contribution of the microbiome. LBP has a central role in the innate immune system by binding to bacterial lipopolysaccharides (LPS) and facilitating the affinity between LPS and CD14, with the subsequent release of various cytokines. Several extracellular matrix proteins could be identified within the top 25 of upregulated genes, including MMP13 (FC = 358.8, p = 1.3E−11), ADAM12 (FC = 175.6, p = 1.4E−12), COL1A1 (FC = 77.1, p = 2.7E−10) and COL5A3 (FC = 32.1, p = 1.7E−12), emphasising the intense tissue remodelling going on in the fistula tract. Despite correcting for the confounding effect of proctitis, the fistula tract expressed higher levels of IL6 (FC = 133.1, p = 3.7E−9), TNF(FC = 14.2, p = 4.8E−5), OSM (FC = 24.3, p = 8.9E−5), IL12p40 (FC = 10.0, p = 8.1E−3), integrin α 4 (FC = 4.5, p = 9.3E−3), integrin β 7(FC = 3.8, p = 4.1E−3) but not IL23p19 (FC = 1.2, p = 0.9). Top downregulated genes were linked to the intestinal epithelium, including KRT19 (FC = −489.1, p = 5.5E−17), KRT8 (FC = −324.0, p = 1.1E−16), CEACAM6 (FC=−515.1, p = 4.5E−16) and MUC2 (FC=−795.4, p = 3.0E−15). Cellular deconvolution identified CD4 memory resting T cells (18.5%), M0 macrophages (17%), M2 macrophages (15.2%), neutrophils (9.2%) and plasma cells (7.5%) as the most abundant cells within the fistula tract.

Conclusion

We molecularly characterised the fistula tract in perianal CD and identified clear biological differences in comparison to the luminal tract, highlighting the potential of new therapeutic targets and cell types driving this debilitating condition.
P065 Metabolic utilisation of propanediol by adherent-invasive E. coli regulates intestinal tissue immunity
Authors: Viladomiu M , Metz M , Lima S , Guo C J , Simpson K , Scherl E , Longman R
Published: January 2020

Background

Our lab has recently shown that adherent-invasive E. coli (AIEC), which is enriched in patients with Crohn’s disease, is sufficient to induce intestinal Th17 cell inflammation. Although AIEC lack pathogenic factors including type III secretion systems, virulence-associated metabolic enzymes, including propanediol dehydratase (PduC), enable AIEC to use fucose-derived propanediol as an alternate carbon source in the gut. We hypothesised that this propanediol utilisation pathway functionally modulates AIEC contribution to intestinal tissue immunity and inflammation.

Methods

We generated pduC-deficient (ΔpduC) and catalytically inactive pduC (pduC*) mutants of a patient-derived, AIEC isolate. Gnotobiotic and genetic mouse models were used to evaluate the cellular and metabolic contribution of pduC to mucosal Th17-cell inflammation.

Results

We found that pduC is enriched in the microbiome and within E. coli genomes in Crohn’s disease patients compared with healthy controls. Mono-colonisation of mice with pduC-deficient AIEC resulted in reduced, inflammatory Th17 cells and attenuated weight loss using a T-cell transfer colitis model. Using mouse models for inducible deletion of CX3CR1+mononuclear phagocytes (MNPs) or IL10-deficiency, we found that MNPs are required for AIEC-mediated induction of Th17 cells and that IL-10 is required to restrain pduC-dependent colitis. Using a catalytically inactive mutant in mono-colonised mice, we determined that PduC metabolic activity was required for this immune phenotype. Cell-free supernatants from WT AIEC (but not pduC-deficient AIEC) promoted ex vivo Th17 cell polarisation and LC-MS of these supernatants defined PduC-dependent metabolites capable of promoting Th17 polarisation.

Conclusion

These data reveal a link between AIEC microbial metabolism and inflammatory Th17 cells with the potential to serve as a therapeutic target for tissue inflammation in the treatment of Crohn’s disease.
P078 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

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 flow-through laser cytofluorimeter (Cytomics FC500, Beckman Coulte). The percentage of monocytes (CD14 + cells) carrying TLR2, TLR4, TLR6 on their surface was assessed. The separation of proteins of colon mucosa was based on technologies of IEF, SDS–PAGE, 2DPAGE (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 < 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 < 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 are 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.
P092 Pathogenic B-cell response in ulcerative colitis that associates with treatment resistance and disease complications
Authors: Uzzan M , Martin J , Keningsberg E , Castro-Dopico T , Huang R , Ungaro R , Cohen B , Rahman A , Rosenstein A , Gurunathan S , Dubinsky M , Clatworthy M , Suarez-Farinas M , Argmann C , Colombel J F , Mehandru S
Published: January 2020

Background

Among the adaptive immune cells, B cells, dominated by IgA producing plasma cells (PC), are critical for intestinal homeostasis. However, the B-cell response remains under studied in ulcerative colitis (UC).

Methods

Using single-cell RNA sequencing (scRNA seq), single-cell IgH sequencing (sc IgH seq), multiparameter flow cytometry (FC), mass cytometry (MC) and immunofluorescent microscopy (IF), we have comprehensively defined the landscape of intestinal and circulating B cells in patients with UC. A Cohort of 88 patients with active UC, 14 patients with quiescent UC and 43 healthy volunteers were recruited. Two validation cohorts comprising of 53 patients with active UC, 59 patients with quiescent UC and 65 healthy volunteers (Validation cohort 1) and 65 patients with active UC, 42 patients with quiescent UC and 36 healthy volunteers (Validation cohort 2) were also studied.

Results

scRNA seq and FC demonstrate a dramatic increase in proliferating (Ki67+) plasmablasts (PB) as well as IgG+ PC in the inflamed colon of UC patients. Additionally, a significant increase in short-lived (CD19+CD45+) PC was observed during colonic inflammation. scIgH seq further demonstrates a significant decrease in somatic hypermutations and the diversity of intestinal PC of patients with active UC. Genes associated with Fc-dependent macrophage signalling were enriched in areas of active disease and were associated with non-response to TNF therapy. In circulation, a significant increase in a4b7+ gut-homing PC was noted that related to disease extent, severity and complications (defined by treatment non-response, hospitalisations and surgery).

Conclusion

Active UC results in an immature, IgG-dominated PB/PC response in inflamed GI tissue and an increase in the frequency of gut-homing PC in circulation that relates to disease activity and complications and may be used to monitor disease course. These data provide novel insights into the pathogenesis of UC with major implications for existing and emerging therapeutic agents.
P097 Depletion of delta-like ligand 4 (Dll4) contributes to the alteration of secretory progenitor differentiation in SAMP1/YitFc mice with Crohn’s disease-like ileitis
Authors: Hong S N , Song J H , Seong G , Kong S M , Shin J B , Eun Ran K , Dong Kyung C , Young-Ho K
Published: January 2020

Background

When the intestinal epithelium would be exposed to harmful substances, mucosal damage can be occurred and followed by epithelial restoration with regenerative epithelium derived from intestinal stem cells (ISCs). Crohn’s disease (CD) is characterised by chronic mucosal damage and ulceration predominantly involved in the ileum. Thus, we hypothesised that epithelial regeneration in CD might be impaired due to the dysfunction of ileal ISCs or its niche.

Methods

SAMP1/YitFcsI (SAMP1) mice are recombinant-inbred mice that spontaneously develop ileitis resembling human CD. Alterations in the crypt-villus structure, epithelial differentiation, organoid formation ability, and niche signalling pathways in the ileum of 10- to 14-week-old SAMP1 mice were analysed and compared with age-matched AKR mice.

Results

In the ileum of SAMP1 mice, the depth of the crypts increased and the surface area of the villi decreased. While the number of ISCs in the ileal crypts did not differ between SAMP1 and AKR mice. The number of Paneth cells decreased and the number of transient amplifying cells increased. The organoid formation ability of the ileal crypts of SAMP1 mice decreased significantly compared with that of AKR mice. RNA-seq revealed a differential gene expression pattern of ileal crypts of SAMP1 mice compared with those of AKR mice. Among the ISCs and niche signalling, the expression of delta-like ligand 4 (Dll4) was significantly decreased and the expression of genes associated with Notch signalling tended to decrease in the ileal crypts of SAMP1 mice.

Conclusion

The characteristic ileitis in SAMP1 mice may be caused by impaired Dll4-Notch signalling.
P116 The occurrence of venous thromboembolisms in paediatric-onset IBD
Authors: Aardoom M , Kemos P , Ruemmele F M , van Ommen C H , Croft N M , de Ridder L
Published: January 2020

Background

Venous thromboembolisms (VTEs) are one of the severe adverse events that may occur in patients with paediatric-onset inflammatory bowel disease (IBD). Based on available literature VTE incidence in the general paediatric population ranges from 0.07 to 0.49 per 10,000 children. Several studies in adults suggest an increased risk of venous thromboembolism in adult IBD patients, but data in paediatric IBD patients is limited. Due to the rarity of these events, international studies are needed to gain information on risk factors for this complication and optimise guidelines. With this study, we aim to assess the incidence and characteristics of VTEs in children with IBD.

Methods

As part of an international prospective safety registry in the PIBD-SETQuality project, paediatric gastroenterologists reply monthly to an electronic survey to indicate whether they have seen one of 10 predetermined complications, including VTE, in an IBD patient <19 years of age. When a VTE is reported, additional information on patient and disease characteristics and previous therapy strategies is collected. In addition, participating physicians annually report the number of new and current PIBD patients under their care, which combined with the reporting period, is the denominator data for this project.

Results

Since October 2016, 150 paediatric gastroenterologists from 26 different countries are participating in the safety registry. In total, 14 cases of VTE were reported for 28,793 patient-years of follow-up. This results in an observed incidence of 4.86 per 10,000 PIBD patients annually (95% CI 2.66–8.16), which is a 10-fold increase compared with the VTE incidence reported in the literature in the general paediatric population. Among cases with available additional information (n = 10) were five cases of intracranial thromboembolism, four upper or lower limbs VTE and one pulmonary embolism, which occurred mainly in children with ulcerative colitis (UC) or IBD unclassified (IBDU) (5/8). Seventy per cent of cases occurred during an exacerbation of IBD, but only 40% during hospitalisation. Physicians reported steroid therapy (n = 4), immobility (n = 1), surgery (n = 1) and the presence of a central venous catheter (n = 1) as risk factors.

Conclusion

This registry has the ability to collect rare but severe complications and patient-at-risk information in paediatric IBD. Our data suggest a 10-fold increase of the VTE incidence in the paediatric IBD population compared with the general paediatric population. Most VTEs were reported in UC/IBD-U patients and occurred during an exacerbation of IBD. Ongoing data collection will help to understand possible causes and gain insight in differences in management and outcomes of these complications per country or region.
P119 Brain function and fatigue in patients with inflammatory bowel disease
Authors: Thomann A , Schmitgen M , Kmuche D , Griebe M , Ebert M , Wolf R C , Reindl W
Published: January 2020

Background

Fatigue is common in patients with inflammatory bowel diseases (IBD). It occurs in up to 80% of patients with active disease, but also a considerable proportion of patients in remission, and significantly affects quality of life. The underlying mechanisms are still poorly understood, and it is still unclear which patients will suffer from fatigue even in luminally quiescent disease. Task-based brain functional studies have examined neural correlates of fatigueability and found changes in the orbitofrontal cortex (OFC), among other regions. To the best our knowledge, the relationship between brain function and fatigue in IBD has not been investigated. This study aimed to examine the association between fatigue and resting-state brain function in remitted IBD patients.

Methods

We obtained resting-state-functional MRI (rs-fMRI) data from 45 IBD patients in stable remission without current steroid or biological therapy and 17 healthy controls (HCs). Fatigue was assessed with Würzburger Erschöpfungsinventar Multiple Sklerose (WEIMuS). Preprocessing of rs-fMRI-data and calculation of amplitude of low frequency fluctuations (ALFF) was performed using the Data Processing Assistant for rs-fMRI. The resulting individual maps were analysed via second-level SPM multiple regression models in patients and HC to test for correlations between ALFF data and WEIMuS scores. Age, gender and mean framewise displacement were included as covariates of no interest and results were displayed at p < 0.001 (peak level) with a threshold of spatial extent (k) according to the expected voxels per cluster estimated by SPM.

Results

Fatigue scores did not differ significantly between patients and controls (mean WEIMuS-scores 17.9 (SD 13.9) vs. 12.3 (SD 16.5), p = .17). Proportions of participants with fatigue scores above the cutoff (>32P.) were nearly identical in patients and HC (8/45 vs. 3/17). In patients, fatigue scores correlated positively with ALFF in the right central operculum and negatively with ALFF in the left OFC and left cerebellum (all p < .001, Figure 1). Fatigue and ALFF in the left cerebellum were also found to correlate in HC.

Conclusion

This study shows fatigue-associated changes in brain activity in several brain regions. The negative association between fatigue and ALFF in the left OFC of IBD patients was not seen in HC, indicating that reduced ALFF in the OFC may represent a neural correlate of IBD-related fatigue. The OFC is thought to be involved in decision-making, which is described to be impaired in many fatigued IBD patients. If the association between fatigue and brain function detected in our study is confirmed in longitudinal IBD studies, these regions could serve as biomarkers when targeting fatigue in IBD.
P141 Scoring endoscopy in pediatric inflammatory bowel disease: a way to improve quality
Authors: Norsa L , Ferrari A , Arrigo S , Bramuzzo M , Deganello Saccomani M , Di Nardo G , Illiceto M T , Miele E , Paci M , Romano C , Romeo E , Daperno M , oliva S
Published: January 2020

Background

The aim of mucosal healing (MH) as a therapeutic target in paediatric inflammatory bowel diseases (IBD) has emphasised the role of the endoscopy. There is a great variability in evaluating mucosal lesions among different operators, especially in paediatric patients. This multicentre prospective study aims to evaluate the interobserver agreement among paediatric endoscopists in using validated endoscopic scores of IBD in children.

Methods

Fifteen videos of follow-up ileocolonoscopies in children with IBD (8 ulcerative colitis –UC-, 7 Crohn’s disease –CD-) were selected from 3 different referral sites in Italy. Eleven paediatric endoscopists from different centres were asked to evaluate all videos as independent and blinded readers. The scoring systems used were ulcerative colitis Endoscopic Index of Severity (UCEIS) for UC and simple endoscopic score for Crohn’s disease (SES-CD) for CD. Kappa statistics and intraclass correlation coefficients were used to measure agreement. Furthermore, an experienced adult gastroenterologist evaluated the same videos and scores them. His results were compared with paediatric endoscopists’ findings.

Results

The median age of the participants was 40 (interquartile range: 6) with a median experience of 12 (14) years in centres with a median number of 140 (230) of paediatric IBDs. Intercluster correlation agreement was 0.298 (95% CI: 0.13–0.55) for UC and 0.266 (0.11–0.52) for CD. When a disease activity categorisation was adopted (remission, moderate, mild and severe) Fleiss’ kappa coefficient was 0.408 (0.29–0.53) for UC and 0.552 (0.43–0.73) for CD (Figure 1). When stratified for item vascular pattern of UC was the most reliable item IC: 0.624 (0.321–0.854). The comparison between paediatric and expert gastroenterologist’s scores is shown in Figure 2. In the multivariate analysis none of the reviewer characteristic affected the readers’ errors.

Conclusion

This pilot multicentre study shows that there is a low level of agreement among paediatric endoscopists in evaluating children with IBDs. Agreement improved after using a disease activity categorisation, with better results for CD. Regardless to experience, all readers showed a low-grade accordance with adult gastroenterologist. According to these findings, the use of scoring systems should be implemented for all paediatric endoscopists. Future specific training programs should be considered to pursue this goal.
P185 A comparison of early response parameters to inform a treat-to-target approach in ulcerative colitis
Authors: Dulai P , Feagan B , Sands B , Lasch K , Lirio R , Feng D
Published: January 2020

Background

We aimed to evaluate early markers of response after vedolizumab (VDZ) induction therapy for their prognostic value in determining the long-term risk of colectomy.

Methods

Post hoc analysis of VDZ-treated patients (n = 588) enrolled in the GEMINI 1 and long-term safety (LTS) clinical trial programs. Rates of colectomy during the 7-year follow-up period were compared according to treatment response status at week 6 (induction) for various definitions of Clinical remission [CR] (rectal bleeding sub-score [RBS] 0 + stool frequency sub-score [SFS] ≤1), Biomarker remission [BR] (faecal calprotectin [FC] ≤50, ≤100, ≤250), or endoscopic improvement [EI] (Mayo endoscopic sub-score [MES] ≤1) and endoscopic remission [ER] (MES 0). Time-to-event with log-rank and Cox proportional hazard analyses were used and presented as hazard ratios (HR) with 95% confidence intervals (CIs).

Results

The risk of colectomy was comparable between UC patients achieving week 6 CR vs. those with persistent symptoms (HR 0.74, 95% CI 0.37–1.48, p = 0.39). The risk of colectomy trended lower in patients achieving week 6 EI (HR 0.55, 95% CI 0.28–1.10, p = 0.08), but was comparable between those achieving an MES of 1 vs. 0 (HR 1.45, 95% CI 0.39–5.48, p = 0.58). The risk of colectomy was significantly lower in patients achieving week 6 BR using an FC cut-off of 250 (HR 0.30, 95% CI 0.13–0.68, p = 0.002) and 100 (HR 0.18, 95% CI 0.04–0.73, p = 0.007) but not with a cut-off of 50 (HR 0.34, 95% CI 0.08–1.41, p = 0.12). The proportion of patients requiring colectomy over a 7-year period was lower among those achieving week 6 BR (FC 250, 3.3%; FC 100, 1.8%) compared with patients who achieved week 6 EI (5.6%), although over one-third of patients who achieved week 6 BR had moderate–severe endoscopic activity (MES 2–3) at week 6.

Conclusion

FC as an early biomarker of response at week 6 may aid in identifying patients who remain at low risk of future colectomy, despite having persistent symptomatic or endoscopic activity. These findings may be limited by overall low rates of colectomy in GEMINI LTS, but can inform considerations for early treat to target approaches.
P197 A snap-shot survey of IBD patients’ at a tertiary centre: a focus on knowledge and willingness to self-manage
Authors: DYALL L , Kamperidis N , Fofaria R , Fenech V , Tyrrell T , Barber S , Shah J , Randall S , Okpeh C , Arebi N
Published: January 2020

Background

Good quality Care in inflammatory bowel disease (IBD) includes the provision of patient-relevant information.1 Data on the level of patient understanding is lacking. We studied patients’ understanding of their IBD condition.

Methods

The IBD team and patient representatives devised a 16-item questionnaire that collected data on baseline demographics and disease-specific characteristics. Answers denoting understanding were scored positively. A sum (Q-sum) of the individual question scores was expressed as median (range).

Results

150 patients were surveyed (49% male). Thirty-nine per cent had ulcerative colitis, 51% Crohn’s Disease and 10% IBD-unclassified. There were: 39 aged <30, 94 aged 30–60, and 15≥60.

The median (range) Qsum was 9 (1–16). Twelve per cent of patients had a Qsum of ³13. The Qsum of patients < 60 years old was 10 (2–16) compared with 7 (1–13) of the ≥60 age group (p = 0.02). Sixty-two per cent of the younger and 60% of the older group perceived themselves as well informed (p = 0.9). The Qsum scores were 12 (2–14), 10 (2–16) and 8.5 (3–14) for patients with disease duration of ≤1 year, 2–15 and ≥15 years respectively (p < 0.05). There was no difference in scores according to disease and medication type.

96% of patients in the <60 age group were willing to access online information compared with 60% in the ≥60 age group (p < 0.01). Younger patients were more confident in recognising symptoms of a flare [86%] compared with those aged ≥60 [60%] (p = 0.01), and were also more likely to understand how their condition may progress compared with older patients (66% vs. 40%, p < 0.05).

Among patients <60 years only 14% of males and 26% of females (p = 0.08) reported having enough information on fertility. Sixty-six per cent of patients on biologic drugs and 53% on non-biologic drugs considered themselves well informed (p = 0.12). Forty-six per cent of the non-biologics cohort also reported low levels of knowledge on self-management of mild flares. Seventy-six per cent of the biologics group and 87% of the non-biologics group expressed willingness for information on self-management (p = 0.1).

Conclusion

Older patients are less well informed, less confident in recognising symptoms of a flare, and less likely to access on-line resources. The level of knowledge inversely correlated with disease duration. This may reflect the heightened recent awareness on the inclusion of patient education in clinical care and availability of resources. There is an interest in receiving information on self-management of mild flares. Education sessions should be personalised according to patient characteristics and objective measures such as PAM score (Insignia Health), implemented to show benefits.

Reference